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Antonio PERSICO

Professore Ordinario
Dipartimento di Scienze Biomediche, Metaboliche e Neuroscienze sede Policlinico

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Pubblicazioni

2023 - A human leukocyte antigen imputation study uncovers possible genetic interplay between gut inflammatory processes and autism spectrum disorders [Articolo su rivista]
Lombardi, L.; Le Clerc, S.; Wu, C. -L.; Bouassida, J.; Boukouaci, W.; Sugusabesan, S.; Richard, J. -R.; Lajnef, M.; Tison, M.; Le Corvoisier, P.; Barau, C.; Banaschewski, T.; Holt, R.; Durston, S.; Persico, A. M.; Oakley, B.; Loth, E.; Buitelaar, J.; Murphy, D.; Leboyer, M.; Zagury, J. -F.; Tamouza, R.
abstract

Autism spectrum disorders (ASD) are neurodevelopmental conditions that are for subsets of individuals, underpinned by dysregulated immune processes, including inflammation, autoimmunity, and dysbiosis. Consequently, the major histocompatibility complex (MHC)-hosted human leukocyte antigen (HLA) has been implicated in ASD risk, although seldom investigated. By utilizing a GWAS performed by the EU-AIMS consortium (LEAP cohort), we compared HLA and MHC genetic variants, single nucleotide polymorphisms (SNP), and haplotypes in ASD individuals, versus typically developing controls. We uncovered six SNPs, namely rs9268528, rs9268542, rs9268556, rs14004, rs9268557, and rs8084 that crossed the Bonferroni threshold, which form the underpinnings of 3 independent genetic pathways/blocks that differentially associate with ASD. Block 1 (rs9268528-G, rs9268542-G, rs9268556-C, and rs14004-A) afforded protection against ASD development, whilst the two remaining blocks, namely rs9268557-T, and rs8084-A, associated with heightened risk. rs8084 and rs14004 mapped to the HLA‐DRA gene, whilst the four other SNPs located in the BTNL2 locus. Different combinations amongst BTNL2 SNPs and HLA amino acid variants or classical alleles were found either to afford protection from or contribute to ASD risk, indicating a genetic interplay between BTNL2 and HLA. Interestingly, the detected variants had transcriptional and/or quantitative traits loci implications. As BTNL2 modulates gastrointestinal homeostasis and the identified HLA alleles regulate the gastrointestinal tract in celiac disease, it is proposed that the data on ASD risk may be linked to genetically regulated gut inflammatory processes. These findings might have implications for the prevention and treatment of ASD, via the targeting of gut-related processes.

2023 - Autism Is Associated With Interindividual Variations of Gray and White Matter Morphology [Articolo su rivista]
Mei, Ting; Forde, Natalie J; Floris, Dorothea L; Dell'Acqua, Flavio; Stones, Richard; Ilioska, Iva; Durston, Sarah; Moessnang, Carolin; Banaschewski, Tobias; Holt, Rosemary J; Baron-Cohen, Simon; Rausch, Annika; Loth, Eva; Oakley, Bethany; Charman, Tony; Ecker, Christine; Murphy, Declan G M; Persico, Antonio M.; incluso nel EU-AIMS LEAP, Group; Beckmann, Christian F; Llera, Alberto; Buitelaar, Jan K
abstract

Background: Although many studies have explored atypicalities in gray matter (GM) and white matter (WM) morphology of autism, most of them relied on unimodal analyses that did not benefit from the likelihood that different imaging modalities may reflect common neurobiology. We aimed to establish brain patterns of modalities that differentiate between individuals with and without autism and explore associations between these brain patterns and clinical measures in the autism group. Methods: We studied 183 individuals with autism and 157 nonautistic individuals (age range, 6-30 years) in a large, deeply phenotyped autism dataset (EU-AIMS LEAP [European Autism Interventions-A Multicentre Study for Developing New Medications Longitudinal European Autism Project]). Linked independent component analysis was used to link all participants' GM volume and WM diffusion tensor images, and group comparisons of modality shared variances were examined. Subsequently, we performed univariate and multivariate brain-behavior correlation analyses to separately explore the relationships between brain patterns and clinical profiles. Results: One multimodal pattern was significantly related to autism. This pattern was primarily associated with GM volume in bilateral insula and frontal, precentral and postcentral, cingulate, and caudate areas and co-occurred with altered WM features in the superior longitudinal fasciculus. The brain-behavior correlation analyses showed a significant multivariate association primarily between brain patterns that involved variation of WM and symptoms of restricted and repetitive behavior in the autism group. Conclusions: Our findings demonstrate the assets of integrated analyses of GM and WM alterations to study the brain mechanisms that underpin autism and show that the complex clinical autism phenotype can be interpreted by brain covariation patterns that are spread across the brain involving both cortical and subcortical areas.

2023 - Commentary: Research status and prospects of acupuncture for autism spectrum disorders [Recensione in Rivista]
Persico, A. M.
abstract

2023 - Diagnostic yield and clinical impact of chromosomal microarray analysis in autism spectrum disorder [Articolo su rivista]
Cucinotta, F.; Lintas, C.; Tomaiuolo, P.; Baccarin, M.; Picinelli, C.; Castronovo, P.; Sacco, R.; Piras, I. S.; Turriziani, L.; Ricciardello, A.; Scattoni, M. L.; Persico, A. M.
abstract

Background: Autism spectrum disorder (ASD) is characterized by high heritability estimates and recurrence rates; its genetic underpinnings are very heterogeneous and include variable combinations of common and rare variants. Array-comparative genomic hybridization (aCGH) offers significant sensitivity for the identification of copy number variants (CNVs), which can act as susceptibility or causal factors for ASD. Methods: The aim of this study was to evaluate both diagnostic yield and clinical impact of aCGH in 329 ASD patients of Italian descent. Results: Pathogenic/likely pathogenic CNVs were identified in 50/329 (15.2%) patients, whereas 89/329 (27.1%) carry variants of uncertain significance. The 10 most enriched gene sets identified by Gene Ontology Enrichment Analysis are primarily involved in neuronal function and synaptic connectivity. In 13/50 (26.0%) patients with pathogenic/likely pathogenic CNVs, the outcome of array-CGH led to the request of 25 additional medical exams which would not have otherwise been prescribed, mainly including brain MRI, EEG, EKG, and/or cardiac ultrasound. A positive outcome was obtained in 12/25 (48.0%) of these additional tests. Conclusions: This study confirms the satisfactory diagnostic yield of aCGH, underscoring its potential for better, more in-depth care of children with autism when genetic results are analyzed also with a focus on patient management.

2023 - Processing of social and monetary rewards in autism spectrum disorders [Articolo su rivista]
Baumeister, Sarah; Moessnang, Carolin; Bast, Nico; Hohmann, Sarah; Aggensteiner, Pascal; Kaiser, Anna; Tillmann, Julian; Goyard, David; Charman, Tony; Ambrosino, Sara; Baron-Cohen, Simon; Beckmann, Christian; Bölte, Sven; Bourgeron, Thomas; Rausch, Annika; Crawley, Daisy; Dell'Acqua, Flavio; Dumas, Guillaume; Durston, Sarah; Ecker, Christine; Floris, Dorothea L; Frouin, Vincent; Hayward, Hannah; Holt, Rosemary; Johnson, Mark H; Jones, Emily J H; Lai, Meng-Chuan; Lombardo, Michael V; Mason, Luke; Oakley, Bethany; Oldehinkel, Marianne; Persico, Antonio M; San José Cáceres, Antonia; Wolfers, Thomas; Loth, Eva; Murphy, Declan G M; Buitelaar, Jan K; Tost, Heike; Meyer-Lindenberg, Andreas; Banaschewski, Tobias; Brandeis, Daniel
abstract

BackgroundReward processing has been proposed to underpin the atypical social feature of autism spectrum disorder (ASD). However, previous neuroimaging studies have yielded inconsistent results regarding the specificity of atypicalities for social reward processing in ASD.AimsUtilising a large sample, we aimed to assess reward processing in response to reward type (social, monetary) and reward phase (anticipation, delivery) in ASD.MethodFunctional magnetic resonance imaging during social and monetary reward anticipation and delivery was performed in 212 individuals with ASD (7.6-30.6 years of age) and 181 typically developing participants (7.6-30.8 years of age).ResultsAcross social and monetary reward anticipation, whole-brain analyses showed hypoactivation of the right ventral striatum in participants with ASD compared with typically developing participants. Further, region of interest analysis across both reward types yielded ASD-related hypoactivation in both the left and right ventral striatum. Across delivery of social and monetary reward, hyperactivation of the ventral striatum in individuals with ASD did not survive correction for multiple comparisons. Dimensional analyses of autism and attention-deficit hyperactivity disorder (ADHD) scores were not significant. In categorical analyses, post hoc comparisons showed that ASD effects were most pronounced in participants with ASD without co-occurring ADHD.ConclusionsOur results do not support current theories linking atypical social interaction in ASD to specific alterations in social reward processing. Instead, they point towards a generalised hypoactivity of ventral striatum in ASD during anticipation of both social and monetary rewards. We suggest this indicates attenuated reward seeking in ASD independent of social content and that elevated ADHD symptoms may attenuate altered reward seeking in ASD.

2023 - RNA sequencing of blood from sex- and age-matched discordant siblings supports immune and transcriptional dysregulation in autism spectrum disorder [Articolo su rivista]
Tomaiuolo, Pasquale; Piras, Ignazio Stefano; Sain, Simona Baghai; Picinelli, Chiara; Baccarin, Marco; Castronovo, Paola; Morelli, Marco J; Lazarevic, Dejan; Scattoni, Maria Luisa; Tonon, Giovanni; Persico, Antonio M
abstract

: Autism spectrum disorder (ASD) is a neurodevelopmental condition with onset in early childhood, still diagnosed only through clinical observation due to the lack of laboratory biomarkers. Early detection strategies would be especially useful in screening high-risk newborn siblings of children already diagnosed with ASD. We performed RNA sequencing on peripheral blood, comparing 27 pairs of ASD children vs their sex- and age-matched unaffected siblings. Differential gene expression profiling, performed applying an unpaired model found two immune genes, EGR1 and IGKV3D-15, significantly upregulated in ASD patients (both p adj = 0.037). Weighted gene correlation network analysis identified 18 co-expressed modules. One of these modules was downregulated among autistic individuals (p = 0.035) and a ROC curve using its eigengene values yielded an AUC of 0.62. Genes in this module are primarily involved in transcriptional control and its hub gene, RACK1, encodes for a signaling protein critical for neurodevelopment and innate immunity, whose expression is influenced by various hormones and known "endocrine disruptors". These results indicate that transcriptomic biomarkers can contribute to the sensitivity of an intra-familial multimarker panel for ASD and provide further evidence that neurodevelopment, innate immunity and transcriptional regulation are key to ASD pathogenesis.

2023 - The future of child and adolescent clinical psychopharmacology: A systematic review of phase 2, 3, or 4 randomized controlled trials of pharmacologic agents without regulatory approval or for unapproved indications [Articolo su rivista]
Cortese, S.; Mcginn, K.; Hojlund, M.; Apter, A.; Arango, C.; Baeza, I.; Banaschewski, T.; Buitelaar, J.; Castro-Fornieles, J.; Coghill, D.; Cohen, D.; Grunblatt, E.; Hoekstra, P. J.; James, A.; Jeppesen, P.; Nagy, P.; Pagsberg, A. K.; Parellada, M.; Persico, A. M.; Purper-Ouakil, D.; Roessner, V.; Santosh, P.; Simonoff, E.; Stevanovic, D.; Stringaris, A.; Vitiello, B.; Walitza, S.; Weizman, A.; Wohlfarth, T.; Wong, I. C. K.; Zalsman, G.; Zuddas, A.; Moreno, C.; Solmi, M.; Correll, C. U.
abstract

We aimed to identify promising novel medications for child and adolescent mental health problems. We systematically searched https://clinicaltrials.gov/ and https://www.clinicaltrialsregister.eu/ (from 01/01/2010–08/23/2022) for phase 2 or 3 randomized controlled trials (RCTs) of medications without regulatory approval in the US, Europe or Asia, including also RCTs of dietary interventions/probiotics. Additionally, we searched phase 4 RCTs of agents targeting unlicensed indications for children/adolescents with mental health disorders. We retrieved 234 ongoing or completed RCTs, including 26 (11%) with positive findings on ≥ 1 primary outcome, 43 (18%) with negative/unavailable results on every primary outcome, and 165 (70%) without publicly available statistical results. The only two compounds with evidence of significant effects that were replicated in ≥ 1 additional RCT without any negative RCTs were dasotraline for attention-deficit/hyperactivity disorder, and carbetocin for hyperphagia in Prader-Willi syndrome. Among other strategies, targeting specific symptom dimensions in samples stratified based on clinical characteristics or established biomarkers may increase chances of success in future development programmes.

2023 - Updated consensus guidelines on the management of Phelan–McDermid syndrome [Articolo su rivista]
Srivastava, S.; Sahin, M.; Buxbaum, J. D.; Berry-Kravis, E.; Soorya, L. V.; Thurm, A.; Bernstein, J. A.; Asante-Otoo, A.; Bennett, W. E.; Betancur, C.; Brickhouse, T. H.; Passos Bueno, M. R.; Chopra, M.; Christensen, C. K.; Cully, J. L.; Dies, K.; Friedman, K.; Gummere, B.; Holder, J. L.; Jimenez-Gomez, A.; Kerins, C. A.; Khan, O.; Kohlenberg, T.; Lacro, R. V.; Levi, L. A.; Levy, T.; Linnehan, D.; Eva, L.; Moshiree, B.; Neumeyer, A.; Paul, S. M.; Phelan, K.; Persico, A.; Rapaport, R.; Rogers, C.; Saland, J.; Sethuram, S.; Shapiro, J.; Tarr, P. I.; White, K. M.; Wickstrom, J.; Williams, K. M.; Winrow, D.; Wishart, B.; Kolevzon, A.
abstract

Phelan–McDermid syndrome (PMS) is a genetic condition caused by SHANK3 haploinsufficiency and characterized by a wide range of neurodevelopmental and systemic manifestations. The first practice parameters for assessment and monitoring in individuals with PMS were published in 2014; recently, knowledge about PMS has grown significantly based on data from longitudinal phenotyping studies and large-scale genotype–phenotype investigations. The objective of these updated clinical management guidelines was to: (1) reflect the latest in knowledge in PMS and (2) provide guidance for clinicians, researchers, and the general community. A taskforce was established with clinical experts in PMS and representatives from the parent community. Experts joined subgroups based on their areas of specialty, including genetics, neurology, neurodevelopment, gastroenterology, primary care, physiatry, nephrology, endocrinology, cardiology, gynecology, and dentistry. Taskforce members convened regularly between 2021 and 2022 and produced specialty-specific guidelines based on iterative feedback and discussion. Taskforce leaders then established consensus within their respective specialty group and harmonized the guidelines. The knowledge gained over the past decade allows for improved guidelines to assess and monitor individuals with PMS. Since there is limited evidence specific to PMS, intervention mostly follows general guidelines for treating individuals with developmental disorders. Significant evidence has been amassed to guide the management of comorbid neuropsychiatric conditions in PMS, albeit mainly from caregiver report and the experience of clinical experts. These updated consensus guidelines on the management of PMS represent an advance for the field and will improve care in the community. Several areas for future research are also highlighted and will contribute to subsequent updates with more refined and specific recommendations as new knowledge accumulates.

2022 - Actionable Genomics in Clinical Practice: Paradigmatic Case Reports of Clinical and Therapeutic Strategies Based upon Genetic Testing [Articolo su rivista]
Butler, M. G.; Moreno-De-luca, D.; Persico, A.
abstract

In clinical settings, the information provided by genetic testing can explain the triggers and processes underlying clinical presentations, such as neurodevelopmental disorders, in up to one third of affected individuals. However, translating this knowledge into better and more personalized clinical management to many appears a distant target. This article presents three paradigmatic cases to exemplify how this translational effort can, at least in some instances, be undertaken today with very positive results: (a) a young girl carrying a chr. 16p11.2 duplication can be screened using targeted exams and undertake therapeutic/preventive interventions related to her genetic diagnosis; (b) a 13-year-old boy with intellectual disability and autism spectrum disorder carries a chr. 11q14.1 deletion, partly spanning the DLG2 gene important for synaptic function, and gained over 20 I.Q. points ostensibly due to carbolithium, prescribed in the absence of affective symptoms, exclusively following the pathophysiology pointed out by the genetic results; (c) a 58-year-old woman carries a COL3A1 gene variant responsible for the vascular form of Ehler–Danlos syndrome with colon rupture. Detection of this variant in six members of her extended family allows for better clinical management of the proband and targeted genetic counselling for family members at risk of this connective tissue disorder. The unprecedented flow of genetic information available today through new technologies, if interpreted in the light of current knowledge in clinical diagnosis and care of those with connective tissue disorders and neurodevelopmental disturbances, in biology and in neuropsychopharmacology, can promote better clinical and pharmacological treatment, disease surveillance, and management provided and incorporated into the clinical setting.

2022 - Depression and Catatonia Associated With Lansoprazole in an Adolescent With Phelan-McDermid Syndrome: A Case Report [Articolo su rivista]
Persico, A.; Ricciardello, A.; Alessandrini, S.; Viola, L.; Bergonzini, P.; Iughetti, L.; Pani, L.
abstract

2022 - Developing Gene-Based Personalised Interventions in Autism Spectrum Disorders [Articolo su rivista]
Freitag, C. M.; Persico, A; Vorstman, J. A. S.
abstract

2022 - Differential Predictors of Response to Early Start Denver Model vs. Early Intensive Behavioral Intervention in Young Children with Autism Spectrum Disorder: A Systematic Review and Meta-Analysis [Articolo su rivista]
Asta, Lisa; Persico, Antonio M
abstract

The effectiveness of early intensive interventions for Autism Spectrum Disorder (ASD) is now well-established, but there continues to be great interindividual variability in treatment response. The purpose of this systematic review is to identify putative predictors of response to two different approaches in behavioral treatment: Early Intensive Behavioral Interventions (EIBI) and the Early Start Denver Model (ESDM). Both are based upon the foundations of Applied Behavioral Analysis (ABA), but the former is more structured and therapist-driven, while the latter is more naturalistic and child-driven. Four databases (EmBase, PubMed, Scopus and WebOfScience) were systematically screened, and an additional search was conducted in the reference lists of relevant articles. Studies were selected if participants were children with ASD aged 12-48 months at intake, receiving either EIBI or ESDM treatment. For each putative predictor, p-values from different studies were combined using Fisher's method. Thirteen studies reporting on EIBI and eleven on ESDM met the inclusion criteria. A higher IQ at intake represents the strongest predictor of positive response to EIBI, while a set of social cognitive skills, including intention to communicate, receptive and expressive language, and attention to faces, most consistently predict response to ESDM. Although more research will be necessary to reach definitive conclusions, these findings begin to shed some light on patient characteristics that are predictive of preferential response to EIBI and ESDM, and may provide clinically useful information to begin personalizing treatment.

2022 - Efficacy and Safety of Q10 Ubiquinol With Vitamins B and E in Neurodevelopmental Disorders: A Retrospective Chart Review [Articolo su rivista]
Cucinotta, F.; Ricciardello, A.; Turriziani, L.; Mancini, A.; Keller, R.; Sacco, R.; Persico, A
abstract

Increased oxidative stress and defective mitochondrial functioning are shared features among many brain disorders. The aim of this study was to verify retrospectively the clinical efficacy and safety of a metabolic support therapy with Q10 ubiquinol, vitamin E and complex-B vitamins in various neurodevelopmental disorders. This retrospective chart review study included 59 patients (mean age 10.1 ± 1.2 y.o., range 2.5–39 years; M:F = 2.47:1), diagnosed with Autism Spectrum Disorder (n = 17), Autism Spectrum Disorder with co-morbid Intellectual Disability (n = 19), Intellectual Disability or Global Developmental Delay (n = 15), Attention-Deficit/Hyperactivity Disorder (n = 3) and Intellectual Disability in Phelan-McDermid syndrome due to chr. 22q13.33 deletion (n = 5). After a minimum of 3 months of therapy, a positive outcome was recorded in 45/59 (76.27%) patients, with Clinical Global Impression—Improvement scores ranging between 1 (“very much improved”) and 3 (“minimally improved”). The most widespread improvements were recorded in cognition (n = 26, 44.1%), adaptative functioning (n = 26, 44.1%) and social motivation (n = 19, 32.2%). Improvement rates differed by diagnosis, being observed most consistently in Phelan-McDermid Syndrome (5/5, 100%), followed by Intellectual Disability/Global Developmental Delay (13/15, 86.7%), Autism Spectrum Disorder with co-morbid Intellectual Disability (15/19, 78.9%), Autism Spectrum Disorder (11/17, 64.7%) and ADHD (1/3, 33.3%). No significant adverse event or side effect leading to treatment discontinuation were recorded. Mild side effects were reported in 18 (30.5%) patients, with the most frequent being increased hyperactivity (9/59, 15.3%). This retrospective chart review suggests that metabolic support therapy with Q10 ubiquinol, vitamin E and complex-B vitamins is well tolerated and produces some improvement in the majority of patients with neurodevelopmental disorders, especially in the presence of intellectual disability. Randomized controlled trials for each single neurodevelopmental disorder are now warranted to conclusively demonstrate the efficacy of these mitochondrial bioenergetic and antioxidant agents and to estimate their therapeutic effect size.

2022 - From Genes to Therapy in Autism Spectrum Disorder [Articolo su rivista]
Vorstman, J. A. S.; Freitag, C. M.; Persico, A
abstract

2022 - Gut mobilization improves behavioral symptoms and modulates urinary p-cresol in chronically constipated autistic children: A prospective study [Articolo su rivista]
Turriziani, L.; Ricciardello, A.; Cucinotta, F.; Bellomo, F.; Turturo, G.; Boncoddo, M.; Mirabelli, S.; Scattoni, M. L.; Rossi, M.; Persico, A.
abstract

Chronic constipation is common among children with ASD and is associated with more severe hyperactivity, anxiety, irritability, and repetitive behaviors. Young autistic children with chronic constipation display higher urinary, and foecal concentrations of p-cresol, an aromatic compound produced by gut bacteria, known to negatively affect brain function. Acute p-cresol administration to BTBR mice enhances anxiety, hyperactivity and stereotypic behaviors, while blunting social interaction. This study was undertaken to prospectively assess the behavioral effects of gut mobilization in young autistic children with chronic constipation, and to verify their possible correlation with urinary p-cresol. To this aim, 21 chronically constipated autistic children 2–8 years old were evaluated before (T0), 1 month (T1), and 6 months (T2) after intestinal mobilization, recording Bristol stool scale scores, urinary p-cresol concentrations, and behavioral scores for social interaction deficits, stereotypic behaviors, anxiety, and hyperactivity. Gut mobilization yielded a progressive and highly significant decrease in all behavioral symptoms over the 6-month study period. Urinary p-cresol levels displayed variable trends not significantly correlated with changes in behavioral parameters, mainly increasing at T1 and decreasing at T2. These results support gut mobilization as a simple strategy to ameliorate ASD symptoms, as well as comorbid anxiety and hyperactivity, in chronically constipated children. Variation in p-cresol absorption seemingly provides limited contributions, if any, to these behavioral changes. Further research will be needed to address the relative role of reduced abdominal discomfort following mobilization, as compared to specific modifications in microbiome composition and in gut bacteria-derived neuroactive compounds.

2022 - In-depth characterization of neuroradiological findings in a large sample of individuals with autism spectrum disorder and controls [Articolo su rivista]
Ambrosino, S.; Elbendary, H.; Lequin, M.; Rijkelijkhuizen, D.; Banaschewski, T.; Baron-Cohen, S.; Bast, N.; Baumeister, S.; Buitelaar, J.; Charman, T.; Crawley, D.; Dell'Acqua, F.; Hayward, H.; Holt, R.; Moessnang, C.; Persico, A; Sacco, R.; San Jose Caceres, A.; Tillmann, J.; Loth, E.; Ecker, C.; Oranje, B.; Murphy, D.; Durston, S.
abstract

Background: Autism spectrum disorder (ASD) is a group of neurodevelopmental conditions associated with quantitative differences in cortical and subcortical brain morphometry. Qualitative assessment of brain morphology provides complementary information on the possible underlying neurobiology. Studies of neuroradiological findings in ASD have rendered mixed results, and await robust replication in a sizable and independent sample. Methods: We systematically and comprehensively assessed neuroradiological findings in a large cohort of participants with ASD and age-matched controls (total N = 620, 348 ASD and 272 controls), including 70 participants with intellectual disability (47 ASD, 23 controls). We developed a comprehensive scoring system, augmented by standardized biometric measures. Results: There was a higher incidence of neuroradiological findings in individuals with ASD (89.4 %) compared to controls (83.8 %, p = .042). Certain findings were also more common in ASD, in particular opercular abnormalities (OR 1.9, 95 % CI 1.3–3.6) and mega cisterna magna (OR 2.4, 95 % CI 1.4–4.0) reached significance when using FDR, whereas increases in macrocephaly (OR 2.0, 95 % CI 1.2–3.2), cranial deformities (OR 2.4, 95 % CI: 1.0–5.8), calvarian / dural thickening (OR 1.5, 95 % CI 1.0–2.3), ventriculomegaly (OR 3.4, 95 % CI 1.3–9.2), and hypoplasia of the corpus callosum (OR 2.7, 95 % CI 1.1–6.3) did not survive this correction. Furthermore, neuroradiological findings were more likely to occur in isolation in controls, whereas they clustered more frequently in ASD. The incidence of neuroradiological findings was higher in individuals with mild intellectual disability (95.7 %), irrespective of ASD diagnosis. Conclusion: There was a subtly higher prevalence of neuroradiological findings in ASD, which did not appear to be specific to the condition. Individual findings or clusters of findings may point towards the neurodevelopmental mechanisms involved in individual cases. As such, clinical MRI assessments may be useful to guide further etiopathological (genetic) investigations, and are potentially valuable to fundamental ASD research.

2022 - Interindividual Differences in Cortical Thickness and Their Genomic Underpinnings in Autism Spectrum Disorder [Articolo su rivista]
Ecker, Christine; Pretzsch, Charlotte M; Bletsch, Anke; Mann, Caroline; Schaefer, Tim; Ambrosino, Sara; Tillmann, Julian; Yousaf, Afsheen; Chiocchetti, Andreas; Lombardo, Michael V; Warrier, Varun; Bast, Nico; Moessnang, Carolin; Baumeister, Sarah; Dell'Acqua, Flavio; Floris, Dorothea L; Zabihi, Mariam; Marquand, Andre; Cliquet, Freddy; Leblond, Claire; Moreau, Clara; Puts, Nick; Banaschewski, Tobias; Jones, Emily J H; Mason, Luke; Bölte, Sven; Meyer-Lindenberg, Andreas; Persico, Antonio M; Durston, Sarah; Baron-Cohen, Simon; Spooren, Will; Loth, Eva; Freitag, Christine M; Charman, Tony; Dumas, Guillaume; Bourgeron, Thomas; Beckmann, Christian F; Buitelaar, Jan K; Murphy, Declan G M
abstract

Objective: Autism spectrum disorder (ASD) is accompanied by highly individualized neuroanatomical deviations that potentially map onto distinct genotypes and clinical phenotypes. This study aimed to link differences in brain anatomy to specific biological pathways to pave the way toward targeted therapeutic interventions. Methods: The authors examined neurodevelopmental differences in cortical thickness and their genomic underpinnings in a large and clinically diverse sample of 360 individuals with ASD and 279 typically developing control subjects (ages 6-30 years) within the EU-AIMS Longitudinal European Autism Project (LEAP). The authors also examined neurodevelopmental differences and their potential pathophysiological mechanisms between clinical ASD subgroups that differed in the severity and pattern of sensory features. Results: In addition to significant between-group differences in "core" ASD brain regions (i.e., fronto-temporal and cingulate regions), individuals with ASD manifested as neuroanatomical outliers within the neurotypical cortical thickness range in a wider neural system, which was enriched for genes known to be implicated in ASD on the genetic and/or transcriptomic level. Within these regions, the individuals' total (i.e., accumulated) degree of neuroanatomical atypicality was signifi-cantly correlated with higher polygenic scores for ASD and other psychiatric conditions, and it scaled with measures of symptom severity. Differences in cortical thickness deviations were also associated with distinct sensory subgroups, especially in brain regions expressing genes involved in excitatory rather than inhibitory neurotransmission. Conclusions: The study findings corroborate the link between macroscopic differences in brain anatomy and the molecular mechanisms underpinning heterogeneity in ASD, and provide future targets for stratification and subtyping.

2022 - Patterns of connectome variability in autism across five functional activation tasks: findings from the LEAP project [Articolo su rivista]
Looden, Tristan; Floris, Dorothea L; Llera, Alberto; Chauvin, Roselyne J; Charman, Tony; Banaschewski, Tobias; Murphy, Declan; Marquand, Andre F; Buitelaar, Jan K; Beckmann, Christian F; Persico, Antonio M; incluso nell'AIMS-2-TRIALS, Group
abstract

Background: Autism spectrum disorder (autism) is a complex neurodevelopmental condition with pronounced behavioral, cognitive, and neural heterogeneities across individuals. Here, our goal was to characterize heterogeneity in autism by identifying patterns of neural diversity as reflected in BOLD fMRI in the way individuals with autism engage with a varied array of cognitive tasks. Methods: All analyses were based on the EU-AIMS/AIMS-2-TRIALS multisite Longitudinal European Autism Project (LEAP) with participants with autism (n = 282) and typically developing (TD) controls (n = 221) between 6 and 30 years of age. We employed a novel task potency approach which combines the unique aspects of both resting state fMRI and task-fMRI to quantify task-induced variations in the functional connectome. Normative modelling was used to map atypicality of features on an individual basis with respect to their distribution in neurotypical control participants. We applied robust out-of-sample canonical correlation analysis (CCA) to relate connectome data to behavioral data. Results: Deviation from the normative ranges of global functional connectivity was greater for individuals with autism compared to TD in each fMRI task paradigm (all tasks p < 0.001). The similarity across individuals of the deviation pattern was significantly increased in autistic relative to TD individuals (p < 0.002). The CCA identified significant and robust brain-behavior covariation between functional connectivity atypicality and autism-related behavioral features. Conclusions: Individuals with autism engage with tasks in a globally atypical way, but the particular spatial pattern of this atypicality is nevertheless similar across tasks. Atypicalities in the tasks originate mostly from prefrontal cortex and default mode network regions, but also speech and auditory networks. We show how sophisticated modeling methods such as task potency and normative modeling can be used toward unravelling complex heterogeneous conditions like autism.

2022 - Qualitative differences in the spatiotemporal brain states supporting configural face processing emerge in adolescence in autism [Articolo su rivista]
Haartsen, R.; Mason, L.; Garces, P.; Gui, A.; Charman, T.; Tillmann, J.; Johnson, M. H.; Buitelaar, J. K.; Loth, E.; Murphy, D.; Jones, E. J. H.; Ahmad, J.; Ambrosino, S.; Banashewski, T.; Baron-Cohen, S.; Bast, N.; Baumeister, S.; Beckmann, C.; Bolte, S.; Bourgeron, T.; Bours, C.; Brandeis, D.; Cornelissen, I.; Crawley, D.; Davidson, C.; Dell' Acqua, F.; Durston, S.; Ecker, C.; Ellis, C.; Faulkner, J.; Hayward, H.; Hipp, J.; Holt, R.; Lai, M. -C.; Leblond, C.; Meyer-Lindenberg, A.; Moessnang, C.; Oakley, B.; O'Dwyer, L.; Persico, A.; Rausch, A.; Sabet, J.; San Jose Caceres, A.; Simonoff, E.; Tost, H.; Rhein, D. V.
abstract

Background: Studying the neural processing of faces can illuminate the mechanisms of compromised social expertise in autism. To resolve a longstanding debate, we examined whether differences in configural face processing in autism are underpinned by quantitative differences in the activation of typical face processing pathways, or the recruitment of non-typical neural systems. Methods: We investigated spatial and temporal characteristics of event-related EEG responses to upright and inverted faces in a large sample of children, adolescents, and adults with and without autism. We examined topographic analyses of variance and global field power to identify group differences in the spatial and temporal response to face inversion. We then examined how quasi-stable spatiotemporal profiles – microstates – are modulated by face orientation and diagnostic group. Results: Upright and inverted faces produced distinct profiles of topography and strength in the topographical analyses. These topographical profiles differed between diagnostic groups in adolescents, but not in children or adults. In the microstate analysis, the autistic group showed differences in the activation strength of normative microstates during early-stage processing at all ages, suggesting consistent quantitative differences in the operation of typical processing pathways; qualitative differences in microstate topographies during late-stage processing became prominent in adults, suggesting the increasing involvement of non-typical neural systems with processing time and over development. Conclusions: These findings suggest that early difficulties with configural face processing may trigger later compensatory processes in autism that emerge in later development.

2022 - Rare coding variation provides insight into the genetic architecture and phenotypic context of autism [Articolo su rivista]
Fu, J. M.; Satterstrom, F. K.; Peng, M.; Brand, H.; Collins, R. L.; Dong, S.; Wamsley, B.; Klei, L.; Wang, L.; Hao, S. P.; Stevens, C. R.; Cusick, C.; Babadi, M.; Banks, E.; Collins, B.; Dodge, S.; Gabriel, S. B.; Gauthier, L.; Lee, S. K.; Liang, L.; Ljungdahl, A.; Mahjani, B.; Sloofman, L.; Smirnov, A. N.; Barbosa, M.; Betancur, C.; Brusco, A.; Chung, B. H. Y.; Cook, E. H.; Cuccaro, M. L.; Domenici, E.; Ferrero, G. B.; Gargus, J. J.; Herman, G. E.; Hertz-Picciotto, I.; Maciel, P.; Manoach, D. S.; Passos-Bueno, M. R.; Persico, A.; Renieri, A.; Sutcliffe, J. S.; Tassone, F.; Trabetti, E.; Campos, G.; Cardaropoli, S.; Carli, D.; Chan, M. C. Y.; Fallerini, C.; Giorgio, E.; Girardi, A. C.; Hansen-Kiss, E.; Lee, S. L.; Lintas, C.; Ludena, Y.; Nguyen, R.; Pavinato, L.; Pericak-Vance, M.; Pessah, I. N.; Schmidt, R. J.; Smith, M.; Costa, C. I. S.; Trajkova, S.; Wang, J. Y. T.; Yu, M. H. C.; Aleksic, B.; Artomov, M.; Benetti, E.; Biscaldi-Schafer, M.; Borglum, A. D.; Carracedo, A.; Chiocchetti, A. G.; Coon, H.; Doan, R. N.; Fernandez-Prieto, M.; Freitag, C. M.; Gerges, S.; Guter, S.; Hougaard, D. M.; Hultman, C. M.; Jacob, S.; Kaartinen, M.; Kolevzon, A.; Kushima, I.; Lehtimaki, T.; Rizzo, C. L.; Maltman, N.; Manara, M.; Meiri, G.; Menashe, I.; Miller, J.; Minshew, N.; Mosconi, M.; Ozaki, N.; Palotie, A.; Parellada, M.; Puura, K.; Reichenberg, A.; Sandin, S.; Scherer, S. W.; Schlitt, S.; Schmitt, L.; Schneider-Momm, K.; Siper, P. M.; Suren, P.; Sweeney, J. A.; Teufel, K.; del Pilar Trelles, M.; Weiss, L. A.; Yuen, R.; Cutler, D. J.; De Rubeis, S.; Buxbaum, J. D.; Daly, M. J.; Devlin, B.; Roeder, K.; Sanders, S. J.; Talkowski, M. E.
abstract

Some individuals with autism spectrum disorder (ASD) carry functional mutations rarely observed in the general population. We explored the genes disrupted by these variants from joint analysis of protein-truncating variants (PTVs), missense variants and copy number variants (CNVs) in a cohort of 63,237 individuals. We discovered 72 genes associated with ASD at false discovery rate (FDR) ≤ 0.001 (185 at FDR ≤ 0.05). De novo PTVs, damaging missense variants and CNVs represented 57.5%, 21.1% and 8.44% of association evidence, while CNVs conferred greatest relative risk. Meta-analysis with cohorts ascertained for developmental delay (DD) (n = 91,605) yielded 373 genes associated with ASD/DD at FDR ≤ 0.001 (664 at FDR ≤ 0.05), some of which differed in relative frequency of mutation between ASD and DD cohorts. The DD-associated genes were enriched in transcriptomes of progenitor and immature neuronal cells, whereas genes showing stronger evidence in ASD were more enriched in maturing neurons and overlapped with schizophrenia-associated genes, emphasizing that these neuropsychiatric disorders may share common pathways to risk.

2022 - Resting state EEG power spectrum and functional connectivity in autism: a cross-sectional analysis [Articolo su rivista]
Garces, P.; Baumeister, S.; Mason, L.; Chatham, C. H.; Holiga, S.; Dukart, J.; Jones, E. J. H.; Banaschewski, T.; Baron-Cohen, S.; Bolte, S.; Buitelaar, J. K.; Durston, S.; Oranje, B.; Persico, A.; Beckmann, C. F.; Bougeron, T.; Dell'Acqua, F.; Ecker, C.; Moessnang, C.; Charman, T.; Tillmann, J.; Murphy, D. G. M.; Johnson, M.; Loth, E.; Brandeis, D.; Hipp, J. F.
abstract

BACKGROUND: Understanding the development of the neuronal circuitry underlying autism spectrum disorder (ASD) is critical to shed light into its etiology and for the development of treatment options. Resting state EEG provides a window into spontaneous local and long-range neuronal synchronization and has been investigated in many ASD studies, but results are inconsistent. Unbiased investigation in large and comprehensive samples focusing on replicability is needed. METHODS: We quantified resting state EEG alpha peak metrics, power spectrum (PS, 2-32 Hz) and functional connectivity (FC) in 411 children, adolescents and adults (n = 212 ASD, n = 199 neurotypicals [NT], all with IQ > 75). We performed analyses in source-space using individual head models derived from the participants' MRIs. We tested for differences in mean and variance between the ASD and NT groups for both PS and FC using linear mixed effects models accounting for age, sex, IQ and site effects. Then, we used machine learning to assess whether a multivariate combination of EEG features could better separate ASD and NT participants. All analyses were embedded within a train-validation approach (70%-30% split). RESULTS: In the training dataset, we found an interaction between age and group for the reactivity to eye opening (p = .042 uncorrected), and a significant but weak multivariate ASD vs. NT classification performance for PS and FC (sensitivity 0.52-0.62, specificity 0.59-0.73). None of these findings replicated significantly in the validation dataset, although the effect size in the validation dataset overlapped with the prediction interval from the training dataset. LIMITATIONS: The statistical power to detect weak effects-of the magnitude of those found in the training dataset-in the validation dataset is small, and we cannot fully conclude on the reproducibility of the training dataset's effects. CONCLUSIONS: This suggests that PS and FC values in ASD and NT have a strong overlap, and that differences between both groups (in both mean and variance) have, at best, a small effect size. Larger studies would be needed to investigate and replicate such potential effects.

2022 - Stratifying the autistic phenotype using electrophysiological indices of social perception [Articolo su rivista]
Mason, L.; Moessnang, C.; Chatham, C.; Ham, L.; Tillmann, J.; Dumas, G.; Ellis, C.; Leblond, C. S.; Cliquet, F.; Bourgeron, T.; Beckmann, C.; Charman, T.; Oakley, B.; Banaschewski, T.; Meyer-Lindenberg, A.; Baron-Cohen, S.; Bolte, S.; Buitelaar, J. K.; Durston, S.; Loth, E.; Oranje, B.; Persico, A.; Dell'Acqua, F.; Ecker, C.; Johnson, M. H.; Murphy, D.; Jones, E. J. H.
abstract

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by difficulties in social communication, but also great heterogeneity. To offer individualized medicine approaches, we need to better target interventions by stratifying autistic people into subgroups with different biological profiles and/or prognoses. We sought to validate neural responses to faces as a potential stratification factor in ASD by measuring neural (electroencephalography) responses to faces (critical in social interaction) in N = 436 children and adults with and without ASD. The speed of early-stage face processing (N170 latency) was on average slower in ASD than in age-matched controls. In addition, N170 latency was associated with responses to faces in the fusiform gyrus, measured with functional magnetic resonance imaging, and polygenic scores for ASD. Within the ASD group, N170 latency predicted change in adaptive socialization skills over an 18-month follow-up period; data-driven clustering identified a subgroup with slower brain responses and poor social prognosis. Use of a distributional data-driven cutoff was associated with predicted improvements of power in simulated clinical trials targeting social functioning. Together, the data provide converging evidence for the utility of the N170 as a stratification factor to identify biologically and prognostically defined subgroups in ASD.

2022 - Urinary Untargeted Metabolic Profile Differentiates Children with Autism from Their Unaffected Siblings [Articolo su rivista]
Timperio, Anna Maria; Gevi, Federica; Cucinotta, Francesca; Ricciardello, Arianna; Turriziani, Laura; Scattoni, Maria Luisa; Persico, Antonio M
abstract

Autism Spectrum Disorder (ASD) encompasses a clinical spectrum of neurodevelopmental conditions that display significant heterogeneity in etiology, symptomatology, and severity. We previously compared 30 young children with idiopathic ASD and 30 unrelated typically-developing controls, detecting an imbalance in several compounds belonging mainly to the metabolism of purines, tryptophan and other amino acids, as well as compounds derived from the intestinal flora, and reduced levels of vitamins B6, B12 and folic acid. The present study describes significant urinary metabolomic differences within 14 pairs, including one child with idiopathic ASD and his/her typically-developing sibling, tightly matched by sex and age to minimize confounding factors, allowing a more reliable identification of the metabolic fingerprint related to ASD. By using a highly sensitive, accurate and unbiased approach, suitable for ensuring broad metabolite detection coverage on human urine, and by applying multivariate statistical analysis, we largely replicate our previous results, demonstrating a significant perturbation of the purine and tryptophan pathways, and further highlight abnormalities in the "phenylalanine, tyrosine and tryptophan" pathway, essentially involving increased phenylalanine and decreased tyrosine levels, as well as enhanced concentrations of bacterial degradation products, including phenylpyruvic acid, phenylacetic acid and 4-ethylphenylsulfate. The outcome of these within-family contrasts consolidates and extends our previous results obtained from unrelated individuals, adding further evidence that these metabolic imbalances may be linked to ASD rather than to environmental differences between cases and controls. It further underscores the excess of some gut microbiota-derived compounds in ASD, which could have diagnostic value in a network model differentiating the metabolome of autistic and unaffected siblings. Finally, it points toward the existence of a "metabolic autism spectrum" distributed as an endophenotype, with unaffected siblings possibly displaying a metabolic profile intermediate between their autistic siblings and unrelated typically-developing controls.

2022 - Yield of array-CGH analysis in Tunisian children with autism spectrum disorder [Articolo su rivista]
Chehbani, F.; Tomaiuolo, P.; Picinelli, C.; Baccarin, M.; Castronovo, P.; Scattoni, M. L.; Gaddour, N.; Persico, A.
abstract

Background: Autism spectrum disorder (ASD) is a neurodevelopmental disorder with strong genetic underpinnings. Microarray-based comparative genomic hybridization (aCGH) technology has been proposed as a first-level test in the genetic diagnosis of ASD and of neurodevelopmental disorders in general. Methods: We performed aCGH on 98 Tunisian children (83 boys and 15 girls) diagnosed with ASD according to DSM-IV criteria. Results: “Pathogenic” or “likely pathogenic” copy number variants (CNVs) were detected in 11 (11.2%) patients, CNVs of “uncertain clinical significance” in 26 (26.5%), “likely benign” or “benign” CNVs were found in 37 (37.8%) and 24 (24.5%) patients, respectively. Gene set enrichment analysis involving genes spanning rare “pathogenic,” “likely pathogenic,” or “uncertain clinical significance” CNVs, as well as SFARI database “autism genes” in common CNVs, detected eight neuronal Gene Ontology classes among the top 10 most significant, including synapse, neuron differentiation, synaptic signaling, neurogenesis, and others. Similar results were obtained performing g: Profiler analysis. Neither transcriptional regulation nor immune pathways reached significance. Conclusions: aCGH confirms its sizable diagnostic yield in a novel sample of autistic children from North Africa. Recruitment of additional families is under way, to verify whether genetic contributions to ASD in the Tunisian population, differently from other ethnic groups, may involve primarily neuronal genes, more than transcriptional regulation and immune-related pathways.

2021 - Atypical Brain Asymmetry in Autism—A Candidate for Clinically Meaningful Stratification [Articolo su rivista]
Floris, D. L.; Wolfers, T.; Zabihi, M.; Holz, N. E.; Zwiers, M. P.; Charman, T.; Tillmann, J.; Ecker, C.; Dell'Acqua, F.; Banaschewski, T.; Moessnang, C.; Baron-Cohen, S.; Holt, R.; Durston, S.; Loth, E.; Murphy, D. G. M.; Marquand, A.; Buitelaar, J. K.; Beckmann, C. F.; Ahmad, J.; Ambrosino, S.; Auyeung, B.; Baumeister, S.; Bolte, S.; Bourgeron, T.; Bours, C.; Brammer, M.; Brandeis, D.; Brogna, C.; de Bruijn, Y.; Chakrabarti, B.; Cornelissen, I.; Crawley, D.; Dumas, G.; Faulkner, J.; Frouin, V.; Garces, P.; Goyard, D.; Ham, L.; Hayward, H.; Hipp, J.; Johnson, M. H.; Jones, E. J. H.; Kundu, P.; Lai, M. -C.; Liogier d'Ardhuy, X.; Lombardo, M. V.; Lythgoe, D. J.; Mandl, R.; Mason, L.; Mennes, M.; Meyer-Lindenberg, A.; Mueller, N.; Oakley, B.; O'Dwyer, L.; Oldehinkel, M.; Oranje, B.; Pandina, G.; Persico, A. M.; Ruggeri, B.; Ruigrok, A.; Sabet, J.; Sacco, R.; San Jose Caceres, A.; Simonoff, E.; Spooren, W.; Toro, R.; Tost, H.; Waldman, J.; Williams, S. C. R.; Wooldridge, C.
abstract

Background: Autism spectrum disorder (“autism”) is a highly heterogeneous neurodevelopmental condition with few effective treatments for core and associated features. To make progress we need to both identify and validate neural markers that help to parse heterogeneity to tailor therapies to specific neurobiological profiles. Atypical hemispheric lateralization is a stable feature across studies in autism, but its potential as a neural stratification marker has not been widely examined. Methods: In order to dissect heterogeneity in lateralization in autism, we used the large EU-AIMS (European Autism Interventions—A Multicentre Study for Developing New Medications) Longitudinal European Autism Project dataset comprising 352 individuals with autism and 233 neurotypical control subjects as well as a replication dataset from ABIDE (Autism Brain Imaging Data Exchange) (513 individuals with autism, 691 neurotypical subjects) using a promising approach that moves beyond mean group comparisons. We derived gray matter voxelwise laterality values for each subject and modeled individual deviations from the normative pattern of brain laterality across age using normative modeling. Results: Individuals with autism had highly individualized patterns of both extreme right- and leftward deviations, particularly in language, motor, and visuospatial regions, associated with symptom severity. Language delay explained most variance in extreme rightward patterns, whereas core autism symptom severity explained most variance in extreme leftward patterns. Follow-up analyses showed that a stepwise pattern emerged, with individuals with autism with language delay showing more pronounced rightward deviations than individuals with autism without language delay. Conclusions: Our analyses corroborate the need for novel (dimensional) approaches to delineate the heterogeneous neuroanatomy in autism and indicate that atypical lateralization may constitute a neurophenotype for clinically meaningful stratification in autism.

2021 - Genotype-phenotype correlation in Phelan-McDermid syndrome: A comprehensive review of chromosome 22q13 deleted genes [Articolo su rivista]
Ricciardello, Arianna; Tomaiuolo, Pasquale; Persico, Antonio M
abstract

Phelan-McDermid syndrome (PMS, OMIM #606232), also known as chromosome 22q13 deletion syndrome, is a rare genetic disorder characterized by intellectual disability, hypotonia, delayed or absent speech, motor impairment, autism spectrum disorder, behavioral anomalies, and minor aspecific dysmorphic features. Haploinsufficiency of SHANK3, due to intragenic deletions or point mutations, is sufficient to cause many neurobehavioral features of PMS. However, several additional genes located within larger 22q13 deletions can contribute to the great interindividual variability observed in the PMS phenotype. This review summarizes the phenotypic contributions predicted for 213 genes distributed along the largest 22q13.2-q13.33 terminal deletion detected in our sample of 63 PMS patients by array-CGH analysis, spanning 9.08 Mb. Genes have been grouped into four categories: (1) genes causing human diseases with an autosomal dominant mechanism, or (2) with an autosomal recessive mechanism; (3) morphogenetically relevant genes, either involved in human diseases with additive co-dominant, polygenic, and/or multifactorial mechanisms, or implicated in animal models but not yet documented in human pathology; (4) protein coding genes either functionally nonrelevant, with unknown function, or pathogenic through mechanisms other than haploinsufficiency; piRNAs, noncoding RNAs, miRNAs, novel transcripts and pseudogenes. Our aim is to understand genotype-phenotype correlations in PMS patients and to provide clinicians with a conceptual framework to promote evidence-based genetic work-ups, clinical assessments, and therapeutic interventions.

2021 - Imbalanced social-communicative and restricted repetitive behavior subtypes of autism spectrum disorder exhibit different neural circuitry [Articolo su rivista]
Bertelsen, N.; Landi, I.; Bethlehem, R. A. I.; Seidlitz, J.; Busuoli, E. M.; Mandelli, V.; Satta, E.; Trakoshis, S.; Auyeung, B.; Kundu, P.; Loth, E.; Dumas, G.; Baumeister, S.; Beckmann, C. F.; Bolte, S.; Bourgeron, T.; Charman, T.; Durston, S.; Ecker, C.; Holt, R. J.; Johnson, M. H.; Jones, E. J. H.; Mason, L.; Meyer-Lindenberg, A.; Moessnang, C.; Oldehinkel, M.; Persico, A.; Tillmann, J.; Williams, S. C. R.; Spooren, W.; Murphy, D. G. M.; Buitelaar, J. K.; Baron-Cohen, S.; Lai, M. -C.; Lombardo, M. V.
abstract

Social-communication (SC) and restricted repetitive behaviors (RRB) are autism diagnostic symptom domains. SC and RRB severity can markedly differ within and between individuals and may be underpinned by different neural circuitry and genetic mechanisms. Modeling SC-RRB balance could help identify how neural circuitry and genetic mechanisms map onto such phenotypic heterogeneity. Here, we developed a phenotypic stratification model that makes highly accurate (97-99%) out-of-sample SC = RRB, SC > RRB, and RRB > SC subtype predictions. Applying this model to resting state fMRI data from the EU-AIMS LEAP dataset (n = 509), we find that while the phenotypic subtypes share many commonalities in terms of intrinsic functional connectivity, they also show replicable differences within some networks compared to a typically-developing group (TD). Specifically, the somatomotor network is hypoconnected with perisylvian circuitry in SC > RRB and visual association circuitry in SC = RRB. The SC = RRB subtype show hyperconnectivity between medial motor and anterior salience circuitry. Genes that are highly expressed within these networks show a differential enrichment pattern with known autism-associated genes, indicating that such circuits are affected by differing autism-associated genomic mechanisms. These results suggest that SC-RRB imbalance subtypes share many commonalities, but also express subtle differences in functional neural circuitry and the genomic underpinnings behind such circuitry.

2021 - Molecular biomarkers to track clinical improvement following an integrative treatment model in autistic toddlers [Articolo su rivista]
Piras, Ignazio S; Manti, Filippo; Costa, Anna; Carone, Valentina; Scalese, Bruna; Talboom, Joshua S; Veronesi, Christian; Tabolacci, Claudio; Persico, Antonio M; Huentelman, Matthew J; Sacco, Roberto; Lintas, Carla
abstract

Objective: Identifying an objective, laboratory-based diagnostic tool (e.g., changes in gene expression), when used in conjunction with disease-specific clinical assessment, could increase the accuracy of the effectiveness of a therapeutic intervention. Methods: We assessed the association between treatment outcome and blood RNA expression before the therapeutic intervention to post-treatment (after one year) of five Autism Spectrum Disorder (ASD) toddlers who underwent an intensive cognitive-behavioral intervention integrated with psychomotor and speech therapy. Results: We found 113 significant Differentially Expressed genes (DEGs) enriched for the nervous system, immune system, and transcription and translation-related pathways. Some of these genes, as MALAT-1, TSPO, and CFL1, appear to be promising candidates. Conclusion: Our findings show that changes in peripheral gene expression could be used in conjunction with clinical scales to monitor a rehabilitation intervention's effectiveness in toddlers affected by ASD. These results need to be validated in a larger cohort.

2021 - Overview of prognostic systems for hepatocellular carcinoma and ITA.LI.CA external validation of MESH and CNLC classifications [Articolo su rivista]
Vitale, A.; Farinati, F.; Finotti, M.; Di Renzo, C.; Brancaccio, G.; Piscaglia, F.; Cabibbo, G.; Caturelli, E.; Missale, G.; Marra, F.; Sacco, R.; Giannini, E. G.; Trevisani, F.; Cillo, U.; Bhoori, S.; Borzio, M.; Burra, P.; Casadei Gardini, A.; Carrai, P.; Conti, F.; Cozzolongo, R.; Cucchetti, A.; D'Ambrosio, R.; Dell'Unto, C.; De Matthaeis, N.; Di Costanzo, G. G.; Di Sandro, S.; Famularo, S.; Foschi, F. G.; Fucilli, F.; Galati, G.; Gambato, M.; Gasbarrini, A.; Giuliante, F.; Ghinolfi, D.; Grieco, A.; Gruttadauria, S.; Guarino, M.; Iavarone, M.; Kostandini, A.; Lai, Q.; Lenci, I.; Levi Sandri, G. V.; Losito, F.; Lupo, L. G.; Marasco, G.; Manzia, T. M.; Mazzocato, S.; Masarone, M.; Melandro, F.; Mescoli, C.; Miele, L.; Morisco, F.; Muley, M.; Nicolini, D.; Pagano, D.; Persico, M.; Pompili, M.; Ponziani, F. R.; Pravisani, R.; Rapaccini, G. L.; Rendina, M.; Renzulli, M.; Romano, F.; Rossi, M.; Rreka, E.; Russo, F. P.; Sangiovanni, A.; Sessa, A.; Simonetti, N.; Sposito, C.; Tortora, R.; Vigano, L.; Vigano, M.; Villa, E.; Vincenzi, V.; Violi, P.; Azzaroli, F.; Brunetto, M. R.; Di Marco, A.; Gasbarrini, A.; Foschi, F. G.; Guarino, M.; Masotto, A.; Mega, A.; Morisco, F.; Nardone, G.; Oliveri, F.; Raimondo, G.; Rapaccini, G. L.; Svegliati Baroni, G.; Vidili, G.; Zoli, M.
abstract

Prognostic assessment in patients with HCC remains an extremely difficult clinical task due to the complexity of this cancer where tumour characteristics interact with degree of liver dysfunction, patient general health status, and a large span of available treatment options. Several prognostic systems have been proposed in the last three decades, both from the Asian and European/North American countries. Prognostic scores, such as the CLIP score and the recent MESH score, have been generated on a solid statistical basis from real life population data, while staging systems, such as the BCLC scheme and the recent CNLC classification, have been created by experts according to recent HCC prognostic evidences from the literature. A third category includes combined prognostic systems that can be used both as prognostic scores and staging systems. A recent example is the ITA.LI.CA prognostic system including either a prognostic score and a simplified staging system. This review focuses first on an overview of the main prognostic systems for HCC classified according to the above three categories, and, second, on a comprehensive description of the methodology required for a correct comparison between different systems in terms of prognostic performance. In this second section the main studies in the literature comparing different prognostic systems are described in detail. Lastly, a formal comparison between the last prognostic systems proposed for each of the above three categories is performed using a large Italian database including 6882 HCC patients in order to concretely apply the comparison rules previously described.

2021 - Preference for biological motion is reduced in ASD: implications for clinical trials and the search for biomarkers [Articolo su rivista]
Mason, L.; Shic, F.; Falck-Ytter, T.; Chakrabarti, B.; Charman, T.; Loth, E.; Tillmann, J.; Banaschewski, T.; Baron-Cohen, S.; Bolte, S.; Buitelaar, J.; Durston, S.; Oranje, B.; Persico, A.; Beckmann, C.; Bougeron, T.; Dell'Acqua, F.; Ecker, C.; Moessnang, C.; Murphy, D.; Johnson, M. H.; Jones, E. J. H.; Ahmad, J.; Ambrosino, S.; Baumeister, S.; Bours, C.; Brammer, M.; Brandeis, D.; Brogna, C.; de Bruijn, Y.; Chatham, C.; Cornelissen, I.; Crawley, D.; Dumas, G.; Faulkner, J.; Frouin, V.; Garces, P.; Goyard, D.; Ham, L.; Hipp, J.; Holt, R.; Lai, M. -C.; D'ardhuy, X. L.; Lombardo, M. V.; Lythgoe, D. J.; Mandl, R.; Marquand, A.; Mennes, M.; Meyer-Lindenberg, A.; Bast, N.; Oakley, B.; O'Dwyer, L.; Oldehinkel, M.; Pandina, G.; Ruggeri, B.; Ruigrok, A.; Sabet, J.; Sacco, R.; Caceres, A. S. J.; Simonoff, E.; Spooren, W.; Toro, R.; Tost, H.; Waldman, J.; Williams, S. C. R.; Wooldridge, C.; Zwiers, M. P.
abstract

Background: The neurocognitive mechanisms underlying autism spectrum disorder (ASD) remain unclear. Progress has been largely hampered by small sample sizes, variable age ranges and resulting inconsistent findings. There is a pressing need for large definitive studies to delineate the nature and extent of key case/control differences to direct research towards fruitful areas for future investigation. Here we focus on perception of biological motion, a promising index of social brain function which may be altered in ASD. In a large sample ranging from childhood to adulthood, we assess whether biological motion preference differs in ASD compared to neurotypical participants (NT), how differences are modulated by age and sex and whether they are associated with dimensional variation in concurrent or later symptomatology. Methods: Eye-tracking data were collected from 486 6-to-30-year-old autistic (N = 282) and non-autistic control (N = 204) participants whilst they viewed 28 trials pairing biological (BM) and control (non-biological, CTRL) motion. Preference for the biological motion stimulus was calculated as (1) proportion looking time difference (BM-CTRL) and (2) peak look duration difference (BM-CTRL). Results: The ASD group showed a present but weaker preference for biological motion than the NT group. The nature of the control stimulus modulated preference for biological motion in both groups. Biological motion preference did not vary with age, gender, or concurrent or prospective social communicative skill within the ASD group, although a lack of clear preference for either stimulus was associated with higher social-communicative symptoms at baseline. Limitations: The paired visual preference we used may underestimate preference for a stimulus in younger and lower IQ individuals. Our ASD group had a lower average IQ by approximately seven points. 18% of our sample was not analysed for various technical and behavioural reasons. Conclusions: Biological motion preference elicits small-to-medium-sized case–control effects, but individual differences do not strongly relate to core social autism associated symptomatology. We interpret this as an autistic difference (as opposed to a deficit) likely manifest in social brain regions. The extent to which this is an innate difference present from birth and central to the autistic phenotype, or the consequence of a life lived with ASD, is unclear.

2021 - Reevaluation of serum arylesterase activity in neurodevelopmental disorders [Articolo su rivista]
Piras, I. S.; Gabriele, S.; Altieri, L.; Lombardi, F.; Sacco, R.; Lintas, C.; Manzi, B.; Curatolo, P.; Nobile, M.; Rigoletto, C.; Molteni, M.; Persico, A. M.
abstract

Organophosphate compounds (OPs) interfere with neurodevelopment and are neurotoxic for humans and animals. They are first biotransformed to the more toxic oxon form, and then hydrolyzed to specific metabolites by the enzyme paraoxonase/arylesterase, encoded by the gene PON1 located on human chr. 7q21.3. In autism spectrum disorder (ASD) and in attention-deficit/hyperactivity disorder (ADHD), a correlation between OP exposure and disease onset has been reported. In this case-control study, we aimed to replicate our previous work showing reduced levels of serum PON1 arylesterase activity in Italian and Caucasian-American ASD samples, and to extend our analysis to other neurodevelopmental disorders, namely ADHD and developmental language disorder (DLD), also known as specific language impairment (SLI). The arylesterase activity, measured using standard spectrophotometric methods, is significantly reduced in the ADHD, and not in the ASD sample compared with the controls. Our previous results seemingly stem from spuriously high arylesterase levels in the former control sample. Finally, genotyping SNPs rs705379 and rs662 using TDI-FP, a significant effect of rs705379 alleles on the serum arylesterase activity is observed in all of the subgroups tested, regardless of diagnosis, as well as a lack of association between PON1 gene polymorphisms and ASD/ADHD susceptibility in the Italian population. In summary, the serum arylesterase activity is reduced in children and adolescents with ADHD, and this reduction is not due to the functional PON1 gene variants assessed in this study. Based on previous literature, it may more likely reflect enhanced oxidative stress than specific genetic underpinnings.

2021 - Saccade dysmetria indicates attenuated visual exploration in autism spectrum disorder [Articolo su rivista]
Bast, N.; Mason, L.; Freitag, C. M.; Smith, T.; Portugal, A. M.; Poustka, L.; Banaschewski, T.; Johnson, M.; Ahmad, J.; Ambrosino, S.; Auyeung, B.; Baron-Cohen, S.; Baumeister, S.; Beckmann, C. F.; Bolte, S.; Bourgeron, T.; Bours, C.; Brammer, M.; Brandeis, D.; Brogna, C.; Bruijn, Y. d.; Buitelaar, J. K.; Chakrabarti, B.; Charman, T.; Cornelissen, I.; Crawley, D.; Dell'Acqua, F.; Dumas, G.; Durston, S.; Ecker, C.; Faulkner, J.; Frouin, V.; Garces, P.; Goyard, D.; Ham, L.; Hayward, H.; Hipp, J.; Holt, R.; Jones, E. J. H.; Kundu, P.; Lai, M. -C.; D'Ardhuy, X. L.; Lombardo, M. V.; Loth, E.; Lythgoe, D. J.; Mandl, R.; Marquand, A.; Mennes, M.; Meyer-Lindenberg, A.; Moessnang, C.; Murphy, D. G. M.; Oakley, B.; O'Dwyer, L.; Oldehinkel, M.; Oranje, B.; Pandina, G.; Persico, A.; Ruggeri, B.; Ruigrok, A.; Sabet, J.; Sacco, R.; San Jose Caceres, A.; Simonoff, E.; Spooren, W.; Tillmann, J.; Toro, R.; Tost, H.; Waldman, J.; Williams, S. C. R.; Wooldridge, C.; Zwiers, M. P.
abstract

Background: Visual exploration in autism spectrum disorder (ASD) is characterized by attenuated social attention. The underlying oculomotor function during visual exploration is understudied, whereas oculomotor function during restricted viewing suggested saccade dysmetria in ASD by altered pontocerebellar motor modulation. Methods: Oculomotor function was recorded using remote eye tracking in 142 ASD participants and 142 matched neurotypical controls during free viewing of naturalistic videos with and without human content. The sample was heterogenous concerning age (6–30 years), cognitive ability (60–140 IQ), and male/female ratio (3:1). Oculomotor function was defined as saccade, fixation, and pupil-dilation features that were compared between groups in linear mixed models. Oculomotor function was investigated as ASD classifier and features were correlated with clinical measures. Results: We observed decreased saccade duration (∆M = −0.50, CI [−0.21, −0.78]) and amplitude (∆M = −0.42, CI [−0.12, −0.72]), which was independent of human video content. We observed null findings concerning fixation and pupil-dilation features (POWER =.81). Oculomotor function is a valid ASD classifier comparable to social attention concerning discriminative power. Within ASD, saccade features correlated with measures of restricted and repetitive behavior. Conclusions: We conclude saccade dysmetria as ASD oculomotor phenotype relevant to visual exploration. Decreased saccade amplitude and duration indicate spatially clustered fixations that attenuate visual exploration and emphasize endogenous over exogenous attention. We propose altered pontocerebellar motor modulation as underlying mechanism that contributes to atypical (oculo-)motor coordination and attention function in ASD.

2021 - The pediatric psychopharmacology of autism spectrum disorder: A systematic review - Part I: The past and the present [Articolo su rivista]
Persico, Antonio M; Ricciardello, Arianna; Lamberti, Marco; Turriziani, Laura; Cucinotta, Francesca; Brogna, Claudia; Vitiello, Benedetto; Arango, Celso
abstract

Autism Spectrum Disorder (ASD) is a severe and lifelong neurodevelopmental disorder, with high social costs and a dramatic burden on the quality of life of patients and family members. Despite its high prevalence, reaching 1/54 children and 1/45 adults in the United States, no pharmacological treatment is still directed to core symptoms of ASD, encompassing social and communication deficits, repetitive behaviors, restricted interests, and abnormal sensory processing. The purpose of this review is to provide an overview of the state-of-the-art of psychopharmacological therapy available today for ASD in children and adolescents, in order to foster best practices and to organize new strategies for future research. To date, atypical antipsychotics such as risperidone and aripiprazole represent the first line of intervention for hyperactivity, impulsivity, agitation, temper outbursts or aggression towards self or others. Tricyclic antidepressants are less prescribed because of uncertain efficacy and important side effects. SSRIs, especially fluoxetine and sertraline, may be effective in treating repetitive behaviors (anxiety and obsessive-compulsive symptoms) and irritability/agitation, while mirtazapine is more helpful with sleep problems. Low doses of buspirone have shown some efficacy on restrictive and repetitive behaviors in combination with behavioral interventions. Stimulants, and to a lesser extent atomoxetine, are effective in reducing hyperactivity, inattention and impulsivity also in comorbid ASD-ADHD, although with somewhat lower efficacy and greater incidence of side effects compared to idiopathic ADHD. Clonidine and guanfacine display some efficacy on hyperactivity and stereotypic behaviors. For several other drugs, case reports and open-label studies suggest possible efficacy, but no randomized controlled trial has yet been performed. Research in the pediatric psychopharmacology of ASD is still faced with at least two major hurdles: (a) Great interindividual variability in clinical response and side effect sensitivity is observed in the ASD population. This low level of predictability would benefit from symptom-specific treatment algorithms and from biomarkers to support drug choice; (b) To this date, no psychoactive drug appears to directly ameliorate core autism symptoms, although some indirect improvement has been reported with several drugs, once the comorbid target symptom is abated.

2020 - Appropriateness of array-CGH in the ADHD clinics: A comparative study [Articolo su rivista]
Baccarin, M.; Picinelli, C.; Tomaiuolo, P.; Castronovo, P.; Costa, A.; Verdecchia, M.; Cannizzaro, C.; Barbieri, G.; Sacco, R.; Persico, A. M.; Lintas, C.
abstract

Attention deficit hyperactivity disorder (ADHD) is one of the most common neurodevelopmental disorder with a worldwide prevalence of about 5%. The disorder is characterized by inattentive, hyperactive and impulsive behavior and is often comorbid with other neuropsychiatric conditions. Array comparative genomic hybridization (array-CGH) testing has been proved to be useful to detect chromosomal aberrations in several neuropsychiatric conditions including autism spectrum disorders (ASD) and intellectual disability (ID). The usefulness of array-CGH in the ADHD clinics is still debated and no conclusive evidence has been reached to date. We performed array-CGH in 98 children and adolescents divided in two similarly sized groups according to the clinical diagnosis: (a) one group diagnosed with ADHD as primary diagnosis; (b) the other group in which ADHD was co-morbid with ASD and/or ID. We detected pathogenetic and likely pathogenetic copy number variants (CNVs) in 12% subjects in which ADHD was co-morbid with autism and/or intellectual disability and in 8.5% subjects diagnosed with ADHD as primary diagnosis. Detection of CNVs of unknown clinical significance was similar in the two groups being 27% and 32%, respectively. Benign and likely benign CNVs accounted for 61% and 59.5% in the first and second group, respectively. Differences in the diagnostic yield were not statistically significant between the two groups (P >.05). Our data strongly suggest that array-CGH (a) is a valuable diagnostic tool to detect clinically significant CNVs in individuals with ADHD even in the absence of comorbidity with ASD and/or ID and (b) should be implemented routinely in the ADHD clinics.

2020 - Autisms [Capitolo/Saggio]
Persico, Antonio; Cucinotta, Francesca; Ricciardello, Arianna; Turriziani, Laura.
abstract

Autism Spectrum Disorder (ASD) is a neurodevelopmental condition with onset in early childhood, characterized by deficits in social interaction and communication, stereotypic behaviors, insistence on sameness, restricted interests and abnormal sensory processing. Its prevalence has risen during the last three decades from 2-5/10,000 to 1:59 children. ASD encompasses a collection of neurodevelopmental conditions sharing similar behavioral features but extremely heterogenous etiopathogenetic underpinnings. This chapter initially reviews the history of the concept of “autism”, the clinical diagnosis, epidemiology, co-morbidities, developmental trajectories and early signs. The pathophysiology of ASD is then described, spanning neuropsychology, neuroanatomy, functional brain imaging and electrophysiology, as well as systemic gastrointestinal and immune dysfunction. The complex etiology of ASD spans from genetic syndroms and non-syndromic autisms due to rare and common variants, to environmental forms and epigenetic contributions. Current evidence-based clinical pharmacological and behavioral intervention paradigms are finally outlined, as well as therapeutic perspectives opened by experimental psychopharmacology and iPSC models.

2020 - Dissecting the phenotypic heterogeneity in sensory features in autism spectrum disorder: A factor mixture modelling approach [Articolo su rivista]
Tillmann, J.; Uljarevic, M.; Crawley, D.; Dumas, G.; Loth, E.; Murphy, D.; Buitelaar, J.; Charman, T.; Ahmad, J.; Ambrosino, S.; Auyeung, B.; Baumeister, S.; Beckmann, C.; Bourgeron, T.; Bours, C.; Brammer, M.; Brandeis, D.; Brogna, C.; De Bruijn, Y.; Chakrabarti, B.; Cornelissen, I.; Acqua, F. D.; Dumas, G.; Ecker, C.; Faulkner, J.; Frouin, V.; Garces, P.; Goyard, D.; Hayward, H.; Hipp, J.; Johnson, M. H.; Jones, E. J. H.; Kundu, P.; Lai, M. -C.; D'Ardhuy, X. L.; Lombardo, M.; Lythgoe, D. J.; Mandl, R.; Mason, L.; Meyer-Lindenberg, A.; Moessnang, C.; Mueller, N.; O'Dwyer, L.; Oldehinkel, M.; Oranje, B.; Pandina, G.; Persico, A. M.; Ruggeri, B.; Ruigrok, A.; Sabet, J.; Sacco, R.; Toro, R.; Tost, H.; Waldman, J.; Williams, S. C. R.; Wooldridge, C.; Zwiers, M. P.
abstract

Background: Heterogeneity in the phenotypic presentation of autism spectrum disorder (ASD) is apparent in the profile and the severity of sensory features. Here, we applied factor mixture modelling (FMM) to test a multidimensional factor model of sensory processing in ASD. We aimed to identify homogeneous sensory subgroups in ASD that differ intrinsically in their severity along continuous factor scores. We also investigated sensory subgroups in relation to clinical variables: sex, age, IQ, social-communication symptoms, restricted and repetitive behaviours, adaptive functioning and symptoms of anxiety and attention-deficit/hyperactivity disorder. Methods: Three hundred thirty-two children and adults with ASD between the ages of 6 and 30 years with IQs varying between 40 and 148 were included. First, three different confirmatory factor models were fit to the 38 items of the Short Sensory Profile (SSP). Then, latent class models (with two-to-six subgroups) were evaluated. The best performing factor model, the 7-factor structure, was subsequently used in two FMMs that varied in the number of subgroups: a two-subgroup, seven-factor model and a three-subgroup and seven-factor model. Results: The 'three-subgroup/seven-factor' FMM was superior to all other models based on different fit criteria. Identified subgroups differed in sensory severity from severe, moderate to low. Accounting for the potential confounding effects of age and IQ, participants in these sensory subgroups had different levels of social-communicative symptoms, restricted and repetitive behaviours, adaptive functioning skills and symptoms of inattention and anxiety. Limitations: Results were derived using a single parent-report measure of sensory features, the SSP, which limits the generalisability of findings. Conclusion: Sensory features can be best described by three homogeneous sensory subgroups that differ in sensory severity gradients along seven continuous factor scores. Identified sensory subgroups were further differentiated by the severity of core and co-occurring symptoms, and level of adaptive functioning, providing novel evidence on the associated clinical correlates of sensory subgroups. These sensory subgroups provide a platform to further interrogate the neurobiological and genetic correlates of altered sensory processing in ASD.

2020 - FARP-1 deletion is associated with lack of response to autism treatment by early start denver model in a multiplex family [Articolo su rivista]
Cucinotta, F.; Ricciardello, A.; Turriziani, L.; Calabrese, G.; Briguglio, M.; Boncoddo, M.; Bellomo, F.; Tomaiuolo, P.; Martines, S.; Bruschetta, M.; La Fauci Belponer, F.; Di Bella, T.; Colombi, C.; Baccarin, M.; Picinelli, C.; Castronovo, P.; Lintas, C.; Sacco, R.; Biederer, T.; Kellam, B.; Scherer, S. W.; Persico, A. M.
abstract

Background: Children with autism spectrum disorder (ASD) display impressive clinical heterogeneity, also involving treatment response. Genetic variants can contribute to explain this large interindividual phenotypic variability. Methods: Array-CGH (a-CGH) and whole genome sequencing (WGS) were performed on a multiplex family with two small children diagnosed with ASD at 17 and 18 months of age. Both brothers received the same naturalistic intervention for one year according to the Early Start Denver Model (ESDM), applied by the same therapists, yielding dramatically different treatment outcomes. Results: The older sibling came out of the autism spectrum, while the younger sibling displayed very little, in any, improvement. This boy was subsequently treated applying a structured Early Intensive Behavioral Intervention paired with Augmentative Alternative Communication, which yielded a partial response within another year. The ESDM nonresponsive child carries a novel maternally inherited 65 Kb deletion at chr. 13q32.2 spanning FARP1. Farp1 is a synaptic scaffolding protein, which plays a significant role in neural plasticity. Conclusion: These results represent a paradigmatic example of the heuristic potential of genetic markers in predicting treatment response and possibly in supporting the targeted prescription of specific early intervention approaches.

2020 - Huntingtin gene CAG repeat size affects autism risk: Family-based and case–control association study [Articolo su rivista]
Piras, I. S.; Picinelli, C.; Iennaco, R.; Baccarin, M.; Castronovo, P.; Tomaiuolo, P.; Cucinotta, F.; Ricciardello, A.; Turriziani, L.; Nanetti, L.; Mariotti, C.; Gellera, C.; Lintas, C.; Sacco, R.; Zuccato, C.; Cattaneo, E.; Persico, A. M.
abstract

The Huntingtin (HTT) gene contains a CAG repeat in exon 1, whose expansion beyond 39 repeats consistently leads to Huntington's disease (HD), whereas normal-to-intermediate alleles seemingly modulate brain structure, function and behavior. The role of the CAG repeat in Autism Spectrum Disorder (ASD) was investigated applying both family-based and case–control association designs, with the SCA3 repeat as a negative control. Significant overtransmission of “long” CAG alleles (≥17 repeats) to autistic children and of “short” alleles (≤16 repeats) to their unaffected siblings (all p < 10−5) was observed in 612 ASD families (548 simplex and 64 multiplex). Surprisingly, both 193 population controls and 1,188 neurological non-HD controls have significantly lower frequencies of “short” CAG alleles compared to 185 unaffected siblings and higher rates of “long” alleles compared to 548 ASD patients from the same families (p <.05–.001). The SCA3 CAG repeat displays no association. “Short” HTT alleles seemingly exert a protective effect from clinically overt autism in families carrying a genetic predisposition for ASD, while “long” alleles may enhance autism risk. Differential penetrance of autism-inducing genetic/epigenetic variants may imply atypical developmental trajectories linked to HTT functions, including excitation/inhibition imbalance, cortical neurogenesis and apoptosis, neuronal migration, synapse formation, connectivity and homeostasis.

2020 - Large-Scale Exome Sequencing Study Implicates Both Developmental and Functional Changes in the Neurobiology of Autism [Articolo su rivista]
Satterstrom, F. K.; Kosmicki, J. A.; Wang, J.; Breen, M. S.; De Rubeis, S.; An, J. -Y.; Peng, M.; Collins, R.; Grove, J.; Klei, L.; Stevens, C.; Reichert, J.; Mulhern, M. S.; Artomov, M.; Gerges, S.; Sheppard, B.; Xu, X.; Bhaduri, A.; Norman, U.; Brand, H.; Schwartz, G.; Nguyen, R.; Guerrero, E. E.; Dias, C.; Aleksic, B.; Anney, R.; Barbosa, M.; Bishop, S.; Brusco, A.; Bybjerg-Grauholm, J.; Carracedo, A.; Chan, M. C. Y.; Chiocchetti, A. G.; Chung, B. H. Y.; Coon, H.; Cuccaro, M. L.; Curro, A.; Dalla Bernardina, B.; Doan, R.; Domenici, E.; Dong, S.; Fallerini, C.; Fernandez-Prieto, M.; Ferrero, G. B.; Freitag, C. M.; Fromer, M.; Gargus, J. J.; Geschwind, D.; Giorgio, E.; Gonzalez-Penas, J.; Guter, S.; Halpern, D.; Hansen-Kiss, E.; He, X.; Herman, G. E.; Hertz-Picciotto, I.; Hougaard, D. M.; Hultman, C. M.; Ionita-Laza, I.; Jacob, S.; Jamison, J.; Jugessur, A.; Kaartinen, M.; Knudsen, G. P.; Kolevzon, A.; Kushima, I.; Lee, S. L.; Lehtimaki, T.; Lim, E. T.; Lintas, C.; Lipkin, W. I.; Lopergolo, D.; Lopes, F.; Ludena, Y.; Maciel, P.; Magnus, P.; Mahjani, B.; Maltman, N.; Manoach, D. S.; Meiri, G.; Menashe, I.; Miller, J.; Minshew, N.; Montenegro, E. M. S.; Moreira, D.; Morrow, E. M.; Mors, O.; Mortensen, P. B.; Mosconi, M.; Muglia, P.; Neale, B. M.; Nordentoft, M.; Ozaki, N.; Palotie, A.; Parellada, M.; Passos-Bueno, M. R.; Pericak-Vance, M.; Persico, A. M.; Pessah, I.; Puura, K.; Reichenberg, A.; Renieri, A.; Riberi, E.; Robinson, E. B.; Samocha, K. E.; Sandin, S.; Santangelo, S. L.; Schellenberg, G.; Scherer, S. W.; Schlitt, S.; Schmidt, R.; Schmitt, L.; Silva, I. M. W.; Singh, T.; Siper, P. M.; Smith, M.; Soares, G.; Stoltenberg, C.; Suren, P.; Susser, E.; Sweeney, J.; Szatmari, P.; Tang, L.; Tassone, F.; Teufel, K.; Trabetti, E.; Trelles, M. D. P.; Walsh, C. A.; Weiss, L. A.; Werge, T.; Werling, D. M.; Wigdor, E. M.; Wilkinson, E.; Willsey, A. J.; Yu, T. W.; Yu, M. H. C.; Yuen, R.; Zachi, E.; Agerbo, E.; Als, T. D.; Appadurai, V.; Baekvad-Hansen, M.; Belliveau, R.; Buil, A.; Carey, C. E.; Cerrato, F.; Chambert, K.; Churchhouse, C.; Dalsgaard, S.; Demontis, D.; Dumont, A.; Goldstein, J.; Hansen, C. S.; Hauberg, M. E.; Hollegaard, M. V.; Howrigan, D. P.; Huang, H.; Maller, J.; Martin, A. R.; Martin, J.; Mattheisen, M.; Moran, J.; Pallesen, J.; Palmer, D. S.; Pedersen, C. B.; Pedersen, M. G.; Poterba, T.; Poulsen, J. B.; Ripke, S.; Schork, A. J.; Thompson, W. K.; Turley, P.; Walters, R. K.; Betancur, C.; Cook, E. H.; Gallagher, L.; Gill, M.; Sutcliffe, J. S.; Thurm, A.; Zwick, M. E.; Borglum, A. D.; State, M. W.; Cicek, A. E.; Talkowski, M. E.; Cutler, D. J.; Devlin, B.; Sanders, S. J.; Roeder, K.; Daly, M. J.; Buxbaum, J. D.
abstract

We present the largest exome sequencing study of autism spectrum disorder (ASD) to date (n = 35,584 total samples, 11,986 with ASD). Using an enhanced analytical framework to integrate de novo and case-control rare variation, we identify 102 risk genes at a false discovery rate of 0.1 or less. Of these genes, 49 show higher frequencies of disruptive de novo variants in individuals ascertained to have severe neurodevelopmental delay, whereas 53 show higher frequencies in individuals ascertained to have ASD; comparing ASD cases with mutations in these groups reveals phenotypic differences. Expressed early in brain development, most risk genes have roles in regulation of gene expression or neuronal communication (i.e., mutations effect neurodevelopmental and neurophysiological changes), and 13 fall within loci recurrently hit by copy number variants. In cells from the human cortex, expression of risk genes is enriched in excitatory and inhibitory neuronal lineages, consistent with multiple paths to an excitatory-inhibitory imbalance underlying ASD.

2020 - P-cresol alters brain dopamine metabolism and exacerbates autism-like behaviors in the BTBR mouse [Articolo su rivista]
Pascucci, T.; Colamartino, M.; Fiori, E.; Sacco, R.; Coviello, A.; Ventura, R.; Puglisi-Allegra, S.; Turriziani, L.; Persico, A. M.
abstract

Background: Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by deficits in social interaction/communication, stereotypic behaviors, restricted interests, and abnormal sensory-processing. Several studies have reported significantly elevated urinary and foecal levels of p-cresol in ASD children, an aromatic compound either of environmental origin or produced by specific gut bacterial strains. Methods: Since p-cresol is a known uremic toxin, able to negatively affect multiple brain functions, the present study was undertaken to assess the effects of a single acute injection of low-or high-dose (1 or 10 mg/kg i.v. respectively) of p-cresol in behavioral and neurochemical phenotypes of BTBR mice, a reliable animal model of human ASD. Results: P-cresol significantly increased anxiety-like behaviors and hyperactivity in the open field, in addition to producing stereotypic behaviors and loss of social preference in BTBR mice. Tissue levels of monoaminergic neurotransmitters and their metabolites unveiled significantly activated dopamine turnover in amygdala as well as in dorsal and ventral striatum after p-cresol administration; no effect was recorded in medial-prefrontal cortex and hippocampus. Conclusion: Our study supports a gene x environment interaction model, whereby p-cresol, acting upon a susceptible genetic background, can acutely induce autism-like behaviors and produce abnormal dopamine metabolism in the reward circuitry.

2020 - Phenotypic spectrum of NRXN1 mono- and bi-allelic deficiency: A systematic review [Articolo su rivista]
Castronovo, P.; Baccarin, M.; Ricciardello, A.; Picinelli, C.; Tomaiuolo, P.; Cucinotta, F.; Frittoli, M.; Lintas, C.; Sacco, R.; Persico, A. M.
abstract

Neurexins are presynaptic cell adhesion molecules critically involved in synaptogenesis and vesicular neurotransmitter release. They are encoded by three genes (NRXN1-3), each yielding a longer alpha (α) and a shorter beta (β) transcript. Deletions spanning the promoter and the initial exons of the NRXN1 gene, located in chromosome 2p16.3, are associated with a variety of neurodevelopmental, psychiatric, neurological and neuropsychological phenotypes. We have performed a systematic review to define (a) the clinical phenotypes most associated with mono-allelic exonic NRXN1 deletions, and (b) the phenotypic features of NRXN1 bi-allelic deficiency due to compound heterozygous deletions/mutations. Clinically, three major conclusions can be drawn: (a) incomplete penetrance and pleiotropy do not allow reliable predictions of clinical outcome following prenatal detection of mono-allelic exonic NRXN1 deletions. Newborn carriers should undergo periodic neuro-behavioral observations for the timely detection of warning signs and the prescription of early behavioral intervention; (b) the presence of additional independent genetic risk factors should always be sought, as they may influence prognosis; (c) children with exonic NRXN1 deletions displaying early-onset, severe psychomotor delay in the context of a Pitt-Hopkins-like syndrome 2 phenotype, should undergo DNA sequencing of the spared NRXN1 allele in search for mutations or very small insertions/deletions.

2020 - Social brain activation during mentalizing in a large autism cohort: The Longitudinal European Autism Project [Articolo su rivista]
Moessnang, C.; Baumeister, S.; Tillmann, J.; Goyard, D.; Charman, T.; Ambrosino, S.; Baron-Cohen, S.; Beckmann, C.; Bolte, S.; Bours, C.; Crawley, D.; Dell'Acqua, F.; Durston, S.; Ecker, C.; Frouin, V.; Hayward, H.; Holt, R.; Johnson, M.; Jones, E.; Lai, M. -C.; Lombardo, M. V.; Mason, L.; Oldenhinkel, M.; Persico, A.; Caceres, A. S. J.; Spooren, W.; Loth, E.; Murphy, D. G. M.; Buitelaar, J. K.; Banaschewski, T.; Brandeis, D.; Tost, H.; Meyer-Lindenberg, A.
abstract

Background: Autism spectrum disorder (ASD) is a neurodevelopmental condition with key deficits in social functioning. It is widely assumed that the biological underpinnings of social impairment are neurofunctional alterations in the "social brain," a neural circuitry involved in inferring the mental state of a social partner. However, previous evidence comes from small-scale studies and findings have been mixed. We therefore carried out the to-date largest study on neural correlates of mentalizing in ASD. Methods: As part of the Longitudinal European Autism Project, we performed functional magnetic resonance imaging at six European sites in a large, well-powered, and deeply phenotyped sample of individuals with ASD (N = 205) and typically developing (TD) individuals (N = 189) aged 6 to 30 years. We presented an animated shapes task to assess and comprehensively characterize social brain activation during mentalizing. We tested for effects of age, diagnosis, and their association with symptom measures, including a continuous measure of autistic traits. Results: We observed robust effects of task. Within the ASD sample, autistic traits were moderately associated with functional activation in one of the key regions of the social brain, the dorsomedial prefrontal cortex. However, there were no significant effects of diagnosis on task performance and no effects of age and diagnosis on social brain responses. Besides a lack of mean group differences, our data provide no evidence for meaningful differences in the distribution of brain response measures. Extensive control analyses suggest that the lack of case-control differences was not due to a variety of potential confounders. Conclusions: Contrary to prior reports, this large-scale study does not support the assumption that altered social brain activation during mentalizing forms a common neural marker of ASD, at least with the paradigm we employed. Yet, autistic individuals show socio-behavioral deficits. Our work therefore highlights the need to interrogate social brain function with other brain measures, such as connectivity and network-based approaches, using other paradigms, or applying complementary analysis approaches to assess individual differences in this heterogeneous condition.

2020 - Temporal Profiles of Social Attention Are Different Across Development in Autistic and Neurotypical People [Articolo su rivista]
Del Bianco, T.; Mason, L.; Charman, T.; Tillman, J.; Loth, E.; Hayward, H.; Shic, F.; Buitelaar, J.; Johnson, M. H.; Jones, E. J. H.; Ahmad, J.; Ambrosino, S.; Banaschewski, T.; Baron-Cohen, S.; Baumeister, S.; Beckmann, C. F.; Bolte, S.; Bourgeron, T.; Bours, C.; Brammer, M.; Brandeis, D.; Brogna, C.; de Bruijn, Y.; Cornelissen, I.; Crawley, D.; Dell'Acqua, F.; Dumas, G.; Durston, S.; Ecker, C.; Faulkner, J.; Frouin, V.; Garces, P.; Goyard, D.; Ham, L.; Hipp, J.; Holt, R.; Lai, M. -C.; D'Ardhuy, X. L.; Lombardo, M. V.; Lythgoe, D. J.; Mandl, R.; Marquand, A.; Mennes, M.; Meyer-Lindenberg, A.; Moessnang, C.; Mueller, N.; Murphy, D. G. M.; Oakley, B.; O'Dwyer, L.; Oldehinkel, M.; Oranje, B.; Pandina, G.; Persico, A. M.; Ruggeri, B.; Ruigrok, A.; Sabet, J.; Sacco, R.; San Jose Caceres, A.; Simonoff, E.; Spooren, W.; Toro, R.; Tost, H.; Waldman, J.; Williams, S. C. R.; Wooldridge, C.; Zwiers, M. P.
abstract

Background: Sociocommunicative difficulties, including abnormalities in eye contact, are core diagnostic features of autism spectrum disorder (ASD). Many studies have used eye tracking to measure reduced attention to faces in autistic people; however, most of this work has not taken advantage of eye-tracking temporal resolution to examine temporal profiles of attention. Methods: We used growth curve analysis to model attention to static social scenes as a function of time in a large (N = 650) sample of autistic participants and neurotypical participants across a wide age range (6–30 years). Results: The model yielded distinct temporal profiles of attention to faces in the groups. Initially, both groups showed a relatively high probability of attending to faces, followed by decline after several seconds. The neurotypical participants, however, were significantly more likely to return their attention to faces in the latter part of each 20-second trial, with increasing probability with age. In contrast, the probability of returning to the face in the autistic participants remained low across development. In participants with ASD, more atypical profiles of attention were associated with lower Vineland Adaptive Behavior Scales communication scores and a higher curvature in one data-driven cluster correlated with symptom severity. Conclusions: These findings show that social attention not only is reduced in ASD, but also differs in its temporal dynamics. The neurotypical participants became more sophisticated in how they deployed their social attention across age, a pattern that was significantly reduced in the participants with ASD, possibly reflecting delayed acquisition of social expertise.

2019 - A mutation in the mitochondrial aspartate/glutamate carrier leads to a more oxidizing intramitochondrial environment and an inflammatory myopathy in Dutch shepherd dogs [Articolo su rivista]
Diane Shelton, G.; Minor, K. M.; Li, K.; Naviaux, J. C.; Monk, J.; Wang, L.; Guzik, E.; Guo, L. T.; Porcelli, V.; Gorgoglione, R.; Lasorsa, F. M.; Leegwater, P. J.; Persico, A. M.; Mickelson, J. R.; Palmieri, L.; Naviaux, R. K.
abstract

Background: Inflammatory myopathies are characterized by infiltration of inflammatory cells into muscle. Typically, immune-mediated disorders such as polymyositis, dermatomyositis and inclusion body myositis are diagnosed. Objective: A small family of dogs with early onset muscle weakness and inflammatory muscle biopsies were investigated for an underlying genetic cause. Methods: Following the histopathological diagnosis of inflammatory myopathy, mutational analysis including whole genome sequencing, functional transport studies of the mutated and wild-type proteins, and metabolomic analysis were performed. Results: Whole genome resequencing identified a pathological variant in the SLC25A12 gene, resulting in a leucine to proline substitution at amino acid 349 in the mitochondrial aspartate-glutamate transporter known as the neuron and muscle specific aspartate glutamate carrier 1 (AGC1). Functionally reconstituting recombinant wild-type and mutant AGC1 into liposomes demonstrated a dramatic decrease in AGC1 transport activity and inability to transfer reducing equivalents from the cytosol into mitochondria. Targeted, broad-spectrum metabolomic analysis from affected and control muscles demonstrated a proinflammatory milieu and strong support for oxidative stress. Conclusions: This study provides the first description of a metabolic mechanism in which ablated mitochondrial glutamate transport markedly reduced the import of reducing equivalents into mitochondria and produced a highly oxidizing and proinflammatory muscle environment and an inflammatory myopathy.

2019 - Altered connectivity between cerebellum, visual, and sensory-motor networks in autism spectrum disorder: results from the EU-AIMS Longitudinal European Autism Project [Articolo su rivista]
Oldehinkel, Marianne; Mennes, Maarten; Marquand, Andre; Charman, Tony; Tillmann, Julian; Ecker, Christine; Dell'Acqua, Flavio; Brandeis, Daniel; Banaschewski, Tobias; Baumeister, Sarah; Moessnang, Carolin; Baron-Cohen, Simon; Holt, Rosemary; Bölte, Sven; Durston, Sarah; Kundu, Prantik; Lombardo, Michael V.; Spooren, Will; Loth, Eva; Murphy, Declan G. M.; Beckmann, Christian F.; Buitelaar, Jan K.; Ahmad, Jumana; Ambrosino, Sara; Auyeung, Bonnie; Bourgeron, Thomas; Bours, Carsten; Brammer, Michael; Brogna, Claudia; de Bruijn, Yvette; Chakrabarti, Bhismadev; Cornelissen, Ineke; Crawley, Daisy; Dumas, Guillaume; Faulkner, Jessica; Frouin, Vincent; Garcés, Pilar; Goyard, David; Ham, Lindsay; Hayward, Hannah; Hipp, Joerg; Johnson, Mark H.; Jones, Emily J. H.; Lai, Meng-Chuan; Liogier D'ardhuy, Xavier; Lythgoe, David J.; Mandl, René; Mason, Luke; Meyer-Lindenberg, Andreas; Mueller, Nico; Oakley, Bethany; O'Dwyer, Laurence; Oranje, Bob; Pandina, Gahan; Persico, Antonio M.; Ruggeri, Barbara; Ruigrok, Amber; Sabet, Jessica; Sacco, Roberto; Cáceres, Antonia San José; Simonoff, Emily; Toro, Roberto; Tost, Heike; Waldman, Jack; Williams, Steve C. R.; Wooldridge, Caroline; Zwiers, Marcel P.
abstract

Background: Resting-state fMRI-based studies on functional connectivity in autism spectrum disorder (ASD) have generated inconsistent results. Interpretation of findings is further hampered by small samples and a focus on a limited number of networks, with networks underlying sensory processing largely under-examined. We aimed to comprehensively characterize ASD-related alterations within and between 20 well-characterized resting-state networks using baseline-data from the EU-AIMS Longitudinal European Autism Project. Methods: Resting-state fMRI data was available for 265 individuals with ASD (7.5-30.3 years; 73.2% male) and 218 typically developing (TD) individuals (6.9-29.8 years; 64.2% male), all with IQ>70. We compared functional connectivity within 20 networks –obtained using independent component analysis– between the ASD and TD group, and related functional connectivity within these networks to continuous (overall) autism trait severity scores derived from the Social Responsiveness Scale-2 across all participants. Furthermore, we investigated case-control differences and autism trait-related alterations in between-network connectivity. Results: Higher autism traits were associated with increased connectivity within salience, medial motor, and orbitofrontal networks. However, we did not replicate previously reported case-control differences within these networks. The between-network analysis did reveal case-control differences showing on average 1) decreased connectivity of the visual association network with somatosensory, medial and lateral motor networks, and 2) increased connectivity of the cerebellum with these sensory and motor networks in ASD compared to TD. Conclusions: We demonstrate ASD-related alterations in within and between-network connectivity. The between-network alterations broadly affect connectivity between cerebellum, visual, and sensorymotor networks, potentially underlying impairments in multisensory and visual-motor integration frequently observed in ASD.

2019 - Deficient Emotional Self-Regulation in Preschoolers With ADHD: Identification, Comorbidity, and Interpersonal Functioning [Articolo su rivista]
Melegari, Maria Grazia; Sacco, Roberto; Manzi, Barbara; Vittori, Elena; Persico, Antonio
abstract

Objective: This study aims to develop an age-adjusted Child Behavior Checklist- (CBCL) and Teacher Report Form (TRF)-based method for the detection of deficient emotional self-regulation (DESR) in preschoolers with ADHD and to assess its incidence, comorbidities, and consequences on interpersonal functioning. Method: Eighty-six ADHD preschoolers and 104 controls were assessed using CBCL, TRF/1½ to 5, Psychiatric Interview With Preschool Age Psychiatric Assessment, Leiter-R, and ADHD rating scales. Results: Greatest sensitivity and specificity were obtained applying slightly lower threshold scores compared with school-age children (CBCL: Anxiety/Depression [A/D] ≥ 59, Attention Problems [AP] ≥ 60, Aggression Behaviors [AB] ≥ 58; TRF: A/D ≥ 59, AP ≥ 60, AB ≥ 60). DESR was detected in 33/86 (38.4%) and in 16/54 (29.6%) ADHD preschoolers versus 2/104 (1.9%) controls using CBCL and TRF, respectively. DESR is associated with significantly greater comorbidity and impairment in interpersonal functioning. Conclusion: Among ADHD preschoolers, DESR (a) requires lower CBCL and TRF threshold scores for detection, compared with school-age children, (b) displays similar incidence rates, and (c) is associated with enhanced psychiatric comorbidity and interpersonal difficulties.

2019 - Evidence that ITGB3 promoter variants increase serotonin blood levels by regulating platelet serotonin transporter trafficking [Articolo su rivista]
Gabriele, Stefano; Canali, Marco; Lintas, Carla; Sacco, Roberto; Tirindelli, Maria Cristina; Ricciardello, Arianna; Persico, Antonio M.
abstract

Elevated serotonin (5-HT) blood levels, the first biomarker identified in autism research, has been consistently found in 20-30% of patients with Autism Spectrum Disorder (ASD). Hyperserotonemia is mainly due to greater 5-HT uptake into platelets, mediated by the 5-HT transporter (SERT) located at the platelet plasma membrane. The protein complex involved in platelet SERT trafficking and externalization includes integrin β3, the beta subunit of the platelet membrane adhesive GP IIb/IIIa. Integrin β3 is encoded by the ITGB3 gene, previously identified as a QTL for 5-HT blood levels in ASD at SNP rs2317385. The present study aims to identify the functional ITGB3 gene variants contributing to hyperserotonemia. ITGB3 gene sequencing in 20 individuals selected on the basis of rs2317385 genotypes defined four haplotypes encompassing six SNPs located in the ITGB3 gene promoter region, all in linkage disequilibrium with rs2317385. Luciferase assays in two hematopoietic cell lines, K-562 and HEL 92.1.7, demonstrate that ITGB3 gene promoter activity is enhanced by the presence of the C allele at rs55827077 specifically during differentiation into megakaryocytes (P<0.01), with modulatory effects by flanking SNPs. This same allele is strongly associated with (a) higher 5-HT blood levels in 176 autistic individuals (P<0.001), (b) greater platelet integrin β3 protein expression (P<0.05), and (c) enhanced SERT trafficking from the cytosol toward the platelet plasma membrane (P=4,05 x 10-11). Our results support rs55827077 as the functional ITGB3 gene promoter variant contributing to elevated 5-HT blood levels in ASD and define a mechanistic chain of events linking ITGB3 to hyperserotonemia.

2019 - Investigating the factors underlying adaptive functioning in autism in the EU-AIMS Longitudinal European Autism Project [Articolo su rivista]
Tillmann, Julian; San José Cáceres, Antonia; Chatham, Chris H.; Crawley, Daisy; Holt, Rosemary; Oakley, Bethany; Banaschewski, Tobias; Baron-Cohen, Simon; Bölte, Sven; Buitelaar, Jan K.; Durston, Sarah; Ham, Lindsay; Loth, Eva; Simonoff, Emily; Spooren, Will; Murphy, Declan G.; Charman, Tony; Ahmad, Jumana; Ambrosino, Sara; Auyeung, Bonnie; Baumeister, Sarah; Beckmann, Christian; Bourgeron, Thomas; Bours, Carsten; Brammer, Michael; Brandeis, Daniel; Brogna, Claudia; de Bruijn, Yvette; Chakrabarti, Bhismadev; Cornelissen, Ineke; Dell’ Acqua, Flavio; Dumas, Guillaume; Ecker, Christine; Faulkner, Jessica; Frouin, Vincent; Garcés, Pilar; Goyard, David; Hayward, Hannah; Hipp, Joerg; Johnson, Mark H.; Jones, Emily J. H.; Kundu, Prantik; Lai, Meng-Chuan; D'ardhuy, Xavier Liogier; Lombardo, Michael; Lythgoe, David J.; Mandl, René; Mason, Luke; Meyer-Lindenberg, Andreas; Moessnang, Carolin; Mueller, Nico; O'Dwyer, Laurence; Oldehinkel, Marianne; Oranje, Bob; Pandina, Gahan; Persico, Antonio M.; Ruggeri, Barbara; Ruigrok, Amber; Sabet, Jessica; Sacco, Roberto; Toro, Roberto; Tost, Heike; Waldman, Jack; Williams, Steve C. R.; Wooldridge, Caroline; Zwiers, Marcel P.
abstract

Scientific Abstract: Individuals with autism spectrum disorder (ASD) exhibit significant impairments in adaptive functioning that impact on their ability to meet the demands of everyday life. A recurrent finding is that there is a pronounced discrepancy between level of cognitive ability and adaptive functioning, and this is particularly prominent among higher-ability individuals. However, the key clinical and demographic associations of these discrepancies remain unclear. This study included a sample of 417 children, adolescents, and adults with ASD as part of the EU-AIMS LEAP cohort. We examined how age, sex, IQ, levels of ASD symptom and autistic trait severity and psychiatric symptomatology are associated with adaptive functioning as measured by the Vineland Adaptive Behavior Scales-Second Edition and IQ adaptive functioning discrepancies. Older age, lower IQ and higher social-communication symptoms were associated with lower adaptive functioning. Results also demonstrate that older age, higher IQ and higher social-communication symptoms are associated with greater IQ-adaptive functioning discrepancy scores. By contrast, sensory ASD symptoms, repetitive and restricted behaviors, as well as symptoms of attention deficit/hyperactivity disorder (ADHD), anxiety and depression, were not associated with adaptive functioning or IQ-adaptive functioning discrepancy scores. These findings suggest that it is the core social communication problems that define ASD that contribute to adaptive function impairments that people with ASD experience. They show for the first time that sensory symptoms, repetitive behavior and associated psychiatric symptoms do not independently contribute to adaptive function impairments. Individuals with ASD require supportive interventions across the lifespan that take account of social-communicative ASD symptom severity.

2019 - The Psychopharmacology of Autism Spectrum Disorder and Rett syndrome [Capitolo/Saggio]
Persico, Antonio; Ricciardello, Arianna; Cucinotta, Francesca
abstract

Autism Spectrum Disorder (ASD) appears in early childhood and is characterized by persistent deficits in communication and social interaction, as well as restricted interests, repetitive behaviors, and unusual sensory issues. ASD can be idiopathic or syndromic, in the latter case representing one of the many manifestations of a genetic disorder, such as Rett Syndrome. Psychopharmacology cannot directly ameliorate core autistic symptoms, but rather aims at treating comorbid disorders, such as ADHD, sleep disturbances, psychomotor agitation and aggressiveness, seizures, and anxiety. Until the 90s, it was mainly based on typical neuroleptics and tricyclic antidepressants, whereas second generation antipsychotics later became first-line drugs for these same indications. In general, selective serotonin reuptake inhibitors have not proven manageable in ASD patients. Psychostimulants are indeed frequently prescribed, but display slightly reduced efficacy and enhanced potential for side effects and non-compliance. Targeted patients benefit from mood stabilizers and antiepileptic drugs. Experimental drug treatments for ASD mainly include oxytocin and vasopressin, bumetanide, sulphorafane, nutritional supplements, memantine and D-cycloserine. Work performed on syndromic forms, represents an important source of information for experimental therapies of ASD. Knowledge of the mechanisms underlying autism in each single patient, is on the verge of paving the path to personalized drug treatments.

2019 - Transcriptome Changes in the Alzheimer's Disease Middle Temporal Gyrus: Importance of RNA Metabolism and Mitochondria-Associated Membrane Genes [Articolo su rivista]
Piras, I. S.; Krate, J.; Delvaux, E.; Nolz, J.; Mastroeni, D. F.; Persico, A. M.; Jepsen, W. M.; Beach, T. G.; Huentelman, M. J.; Coleman, P. D.; Combs, C.
abstract

We used Illumina Human HT-12 v4 arrays to compare RNA expression of middle temporal gyrus (MTG; BA21) in Alzheimer's disease (AD = 97) and non-demented controls (ND = 98). A total of 938 transcripts were highly differentially expressed (adj p < 0.01; log2 FC ≥ |0.500|, with 411 overexpressed and 527 underexpressed in AD. Our results correlated with expression profiling in neurons from AD and ND obtained by laser capture microscopy in MTG from an independent dataset (log2 FC correlation: r = 0.504; p = 2.2e-16). Additionally, selected effects were validated by qPCR. ANOVA analysis yielded no difference between genders in response to AD, but some gender specific genes were detected (e.g., IL8 and AGRN in males, and HSPH1 and GRM1 in females). Several transcripts were associated with Braak staging (e.g., AEBP1 and DNALI1), antemortem MMSE (e.g., AEBP1 and GFAP), and tangle density (e.g., RNU1G2, and DNALI1). At the pathway level, we detected enrichment of synaptic vesicle processes and GABAergic transmission genes. Finally, applying the Weighted Correlation Network Analysis, we identified four expression modules enriched for neuronal and synaptic genes, mitochondria-associated membrane, chemical stimulus and olfactory receptor and non-coding RNA metabolism genes. Our results represent an extensive description of MTG mRNA profiling in a large sample of AD and ND. These data provide a list of genes associated with AD, and correlated to neurofibrillary tangles density. In addition, these data emphasize the importance of mitochondrial membranes and transcripts related to olfactory receptors in AD.

2018 - An interstitial 17q11.2 de novo deletion involving the CDK5R1 gene in a high-functioning autistic patient [Articolo su rivista]
Lintas, Carla; Sacco, Roberto; Tabolacci, Claudio; Brogna, Claudia; Canali, Marco; Picinelli, Chiara; Tomaiuolo, Pasquale; Castronovo, Paola; Baccarin, Marco; Persico, Antonio M.
abstract

We describe a 32-year-old male patient diagnosed with high-functioning autism spectrum disorder carrying a de novo 196-kb interstitial deletion at chromosome 17q11.2. The deletion was detected by array CGH (180K Agilent) and confirmed by quantitative PCR on genomic DNA. The deleted region spans the entire PSMD11 and CDK5R1 genes and partially the MYO1D gene. The CDK5R1 gene encodes for a regulatory subunit of the cyclin-dependent kinase 5 responsible for its brain-specific activation. This gene has been previously associated with intellectual disability in humans. A reduction in CDK5R1 transcript was detected, consistent with the genomic deletion. Based on the functional role of CDK5R1 , this gene appears as the best candidate to explain the clinical phenotype of our patient, whose neuropsychological profile has more resemblance with some of the higher brain function anomalies recently described in the CreERp35 conditional knockout mouse model than previously described patients with intellectual disability.

2018 - Continuità tra psichiatria dell'età evolutiva e psichiatria dell'adulto [Capitolo/Saggio]
Persico, Antonio M.; Nicola, Buonomo
abstract

In Italia, l’esistenza della Neuropsichiatria Infantile e della Psichiatria dell’Adulto ha creato una divisione che vede nei 18 anni un momento di snodo spesso fonte di disorientamento per le famiglie e di difficoltà gestionali-organizzative per i sistemi familiari. Dapprima vengono analizzate le caratteristiche antropologiche delle quattro fasi della vita (infanzia, adolescenza, vita adulta e senescenza). Poi ci si sofferma su due paradigmi di proiezione nella vita adulta di patologie comunemente osservate in età infantile, ossia la disregolazione emotiva, assai frequente nel Disturbo da Deficit di Attenzione con Iperattività, nei disturbi d’ansia e nel disturbo borderline di personalità, e in secondo luogo le somiglianze/differenze tra schizoidia e disturbo autistico ad alto funzionamento, così come si osservano nel bambino e poi evolvono nel giovane adulto. Una corretta distinzione diagnostica tra disturbo schizoide di personalità e sindrome di Asperger nell’adulto consente anche una terapia più appropriata e non può prescindere da una valutazione di quella che è stata anche la traiettoria di sviluppo infantile ed adolescenziale del paziente.

2018 - EU-AIMS Longitudinal European Autism Project (LEAP): the autism twin cohort [Articolo su rivista]
Isaksson, J; Tammimies, K; Neufeld, J; Cauvet, É; Lundin, K; Buitelaar, Jk; Loth, E; Murphy, Dgm; Spooren, W; Bölte, S; Persico, Am; EU-AIMS LEAP, Group.
abstract

EU-AIMS is the largest European research program aiming to identify stratification biomarkers and novel interventions for autism spectrum disorder (ASD). Within the program, the Longitudinal European Autism Project (LEAP) has recruited and comprehensively phenotyped a rare sample of 76 monozygotic and dizygotic twins, discordant, or concordant for ASD plus 30 typically developing twins. The aim of this letter is to complete previous descriptions of the LEAP case-control sample, clinically characterize, and investigate the suitability of the sample for ASD twin-control analyses purposes and share some 'lessons learnt.' Among the twins, a diagnosis of ASD is associated with increased symptom levels of ADHD, higher rates of intellectual disability, and lower family income. For the future, we conclude that the LEAP twin cohort offers multiple options for analyses of genetic and shared and non-shared environmental factors to generate new hypotheses for the larger cohort of LEAP singletons, but particularly cross-validate and refine evidence from it.

2018 - Evaluating Sex and Age Differences in ADI-R and ADOS Scores in a Large European Multi-site Sample of Individuals with Autism Spectrum Disorder [Articolo su rivista]
Tillmann, J.; Ashwood, K.; Absoud, M.; Bölte, S.; Bonnet-Brilhault, F.; Buitelaar, J. K.; Calderoni, S.; Calvo, R.; Canal-Bedia, R.; Canitano, R.; de Bildt, A.; Gomot, M.; Hoekstra, P. J.; Kaale, A.; Mcconachie, H.; Murphy, D. G.; Narzisi, A.; Oosterling, I.; Pejovic-Milovancevic, M.; Persico, A. M.; Puig, O.; Roeyers, H.; Rommelse, N.; Sacco, R.; Scandurra, V.; Stanfield, A. C.; Zander, E.; Charman, T.
abstract

Research on sex-related differences in Autism Spectrum Disorder (ASD) has been impeded by small samples. We pooled 28 datasets from 18 sites across nine European countries to examine sex differences in the ASD phenotype on the ADI-R (376 females, 1763 males) and ADOS (233 females, 1187 males). On the ADI-R, early childhood restricted and repetitive behaviours were lower in females than males, alongside comparable levels of social interaction and communication difficulties in females and males. Current ADI-R and ADOS scores showed no sex differences for ASD severity. There were lower socio-communicative symptoms in older compared to younger individuals. This large European ASD sample adds to the literature on sex and age variations of ASD symptomatology.

2018 - Il Parent Training in età evolutiva [Capitolo/Saggio]
Persico, Antonio M.; Pironti, Erica; Laura, Turriziani
abstract

Si riassume la storia del parent training. Viene esaminato il ruolo del parent training all’interno del progetto terapeutico in Neuropsichiatria Infantile. Le indicazioni principali al parent-training sono l’ADHD e i disturbi esternalizzanti, il disturbo di spettro autistico, i disturbi specifici del linguaggio, enuresi ed encopresi. Vengono presentati alcuni programmi strutturati di parent training, quali “Incredible Years”, il Parent Management Training – The Oregon Model, la Parent Child Interaction Therapy, il Triple-P, il parent training cognitivo-comportamentale, i Cognitive Emotional Relational Groups di Cornoldi e Ferruzza, i programmi per il ritardo del linguaggio espressivo: “It takes two to talk” e “Interact”. Si descrive infine un paradigma clinico di parent training, che coniuga elementi comportamentali per modificare comportamenti problematici ad elementi sistemico-relazionali che mirano ad equilibrare i rapporti intrafamiliari in modo da aumentare l’efficacia dell’intervento genitoriale sui figli con disturbi neuropsichiatrici infantili.

2018 - Il disturbo di spettro autistico [Capitolo/Saggio]
Persico, Antonio M.
abstract

Il Disturbo di Spettro Autistico è una patologia del neurosviluppo caratterizzata da deficit sociocomunicativi, omportamenti stereotipati, interessi ristretti, rigidità cognitiva e disturbi sensoriali, a esordio precoce nell’infanzia e generalmente perdurante per tutta la vita dell’individuo. Colpisce circa 1:50-100 bambini con un aumento esponenziale di prevalenze negli ultimi 20 anni. Viene qui trattata la storia di questa patologia, identificata da Leo Kanner nel 1943, la diagnosi clinica, le co-morbidità, le traiettorie di esordio della malattia nel bambino molto piccolo, l’epidemiologia, la neuropatologia e la neuroanatomia, la neurofisiologia ed il brain imaging funzionale, le funzioni neuropsicologiche compromesse e le relative conseguenze, gli approfondimenti psicodiagnostici e medici, le anomalie gastrointestinali ed immunologiche, l’eziopatogenesi genetica e ambientale, la prognosi, la terapia farmacologica e riabilitativa. Il capitolo mira quindi a fornire una visione completa di questa patologia dagli aspetti scientifici di base fino al disegno del progetto terapeutico-riabilitativo.

2018 - L'anamnesi in neuropsichiatria Infantile [Capitolo/Saggio]
Persico, Antonio M.
abstract

La raccolta dell’anamnesi rappresenta un momento fondante dell’atto medico. In Neuropsichiatria Infantile e dell’Adolescenza, la raccolta di informazioni deve, se possibile, coinvolgere ambedue i genitori, in modo da garantire la massima informatività. Vengono qui riassunti i punti fondamentali da esplorare, relativamente alla vita prenatale (storia di aborti spontanei, difficoltà al concepimento, andamento della gravidanza, parto), la prima infanzia (alimentazione, tappe dello sviluppo psicomotorio, parametri auxometrici, linguaggio), anamnesi patologica specifica, anamnesi scolastica, altre funzioni (attenzione, autocontrollo, oppositività e aggressività, sensorialità, coordinamento motorio, stereotipie e manierismi, adattamento o rigidità degli schemi comportamentali, ritmo sonno-veglia, oralità, dentizione, alimentazione e alvo), comorbidità organiche, interventi riabilitativi, approfondimenti psicometrici e risultato dei test già effettuati, ricoveri, esami ed approfondimenti medici già effettuati, anamnesi familiare. Per ognuno di questi ambiti vengono forniti gli elementi di conoscenza che possono consentire un’anamnesi completa ed esaustiva.

2018 - La valutazione genetica in neuropsichiatria infantile [Capitolo/Saggio]
Persico, Antonio M.; Lintas, Carla
abstract

La valutazione neurogenetica costituisce, insieme alla valutazione psicodiagnostica, elettrofisiologica ed all’imaging cerebrale, una parte fondamentale dell’iter clinico necessario per un corretto inquadramento di moltissimi pazienti in Neuropsichiatria Infantile e dell’Adolescenza. Partendo da un sommario delle diverse fasi del neurosviluppo e dall’esame clinico dismorfologico, si procede nell’analizzare le principali tecniche diagnostiche in uso nella neurogenetica clinica (cariotipo, Fluorescent in Situ Hybridization, fragment analysis, array-CGH, sequenziamento di Sanger e di seconda generazione), per poi applicare questo approccio diagnostico ai disturbi del neurosviluppo, fornendo una flow chart procedurale applicabile nella pratica clinica.

2018 - Le droghe legali:alcol e nicotina nell'adolescenza [Capitolo/Saggio]
Persico, Antonio M.; Lamberti, Marco
abstract

Si descrivono dapprima le caratteristiche generali del sistema nervoso centrale durante l’adolescenza, che promuovendo l’impulsività predispongono alla sperimentazione con sostanze d’abuso. Si analizzano quindi gli effetti dell’alcol e della nicotina, il loro potere drogastico, i loro effetti sul sistema nervoso e sull’organismo del giovane, nonché la loro farmacocinetica. Si analizzano anche la sindrome feto-alcolica e gli effetti dell’esposizione fetale al fumo di sigaretta.

2018 - Le tossicodipendenze: meccanismi neurobiologici [Capitolo/Saggio]
Persico, Antonio M.; Lamberti, Marco
abstract

Questo capitolo descrive i circuiti neurali ed i meccanismi neurotrasmettitoriali coinvolti nel “ciclo delle dipendenze” da sostanze o da comportamenti uncinanti (gioco d’azzardo, videogiochi, smartphone, ecc), partendo dal consumo sporadico, all’abuso, fino alla dipendenza. Viene fornito un “glossario” dei fenomeni più rilevanti che si riscontrano nei fenomeni di dipendenza. I tre ambiti del ciclo dell’addiction, ossia (a) l’intossicazione acuta/abbuffata (“binge”), (b) l’ astinenza/disforia e (c) la preoccupazione/desiderio anticipatorio (“craving”)”, vengono analizzati attraverso il ricorso a modelli animali. Si giunge quindi fino a delineare i fenomeni di plasticità sinaptica e neurale che, a livello soprattutto dei circuiti dopaminergici mesolimbici e mesocorticali, sottende all’instaurarsi di comportamenti di dipendenza e disforia astinenziale spesso misconosciuta nei bambini e negli adolescenti.

2018 - Manuale di Neuropsichiatria Infantile e dell'Adolescenza [Curatela]
Persico, a. m.
abstract

Il Manuale copre tutte le tematiche che tradizionalmente rappresentano “il cuore” della disciplina. Si apre con un capitolo sulla Storia della neuropsichiatria infantile italiana e straniera. Seguono tre sezioni sullo sviluppo tipico, sul processo diagnostico e sui fattori di rischio. Vengono poi analizzati in altrettante sezioni: Disturbi esternalizzanti, Disturbi dell’umore nell’infanzia e nell’adolescenza, Disturbi d’ansia e disturbi somatoformi, Disturbi del controllo sfinterico, Tossicodipendenze e abuso di sostanze, Disturbi del comportamento alimentare, Le cefalee, Epilessie, Disturbi del movimento, Deficit sensoriali, Danni cerebrali su base ipossica, emodinamica, Malattie metaboliche, Sindromi malformative e/o tumorali, Patologie neuromuscolari, Malattie autoimmuni e internistiche, Disturbi del sonno e le parasonnie. Infine segue un’ampia sezione sulle terapie e si conclude con una sezione relativa agli aspetti organizzativi, etici e legali.

2018 - Neurofisiologia ed endocrinologia del comportamento alimentare [Capitolo/Saggio]
Persico, Antonio M.; Galati, Cecilia
abstract

La regolazione del bilancio energetico si fonda su un cross-talk tra fattori periferici e sistema nervoso centrale che da questi fattori viene informato sull’equilibrio anabolismo/catabolismo momento per momento. I fattori biochimici analizzati includono la colecistochinina, il glucagon-like peptide 1, il peptide YY, l’insulina, la leptina e la grelina. Vengono quindi descritti i meccanismi ipotalamici di controllo della fame e della sazietà, nonchè l’influenza esercitata dai sistemi serotoninergico ed endocannabinoide. Infine si riassumono le modificazioni dei livelli dei mediatori biochimici della fame e della sazietà nei disturbi del comportamento alimentare, imostrando che quasi tutti rappresentano conseguenze omeostatiche indotte da uno squilibrio metabolico e non cause dello stesso.

2018 - Prefazione al Manuale di Neuropsichiatria Infantile e dell'Adolescenza [Prefazione o Postfazione]
Persico, Antonio M.
abstract

2018 - Terapia farmacologica personalizzata: basi genetiche e meccanismi d'azione [Capitolo/Saggio]
Persico, Antonio M.; Ricciardello, Arianna
abstract

Questo capitolo descrive le strategie che la ricerca farmacologica attuale sta utilizzando per passare da una farmacologia generica dei disturbi del neurosviluppo ad una farmacologia mirata e personalizzata a correggere specifici meccanismi nel singolo paziente. Dapprima si descrivono i biomarcatori e gli endofenotipi. Poi si passa a descrivere i farmaci in fase di studio per gli autismi sindromici (X-fragile, sindrome di Rett, sclerosi uberosa, PTEN, neurofibromatosi, sindrome di Phelan-McDermid). Infine si prendono in esame i farmaci attualmente in sperimentazione per l’autismo idiopatico, quali ossitocina, vasopressina, bumetanide, sulforafano, farmaci glutammatergici (memantina e D-cicloserina), arbaclofene, Q10 ubiquinolo e L-carnitina, farmaci antiossidanti (N-acetilcisteina, carnosina, vitamina B12, acido folinico, metilB12, glutatione), farmaci attivi sulla neuroinfiammazione e sull’autoimmunità (minociclina, luteolina, quercitina, acidi grassi omega-3), probiotici. Infine si presenta il modello sperimentale delle cellule staminali pluripotenti, come esempio di personalizzazione in vitro prima di somministrare un farmaco in vivo.

2018 - Terapie psicofarmacologiche nel disturbo dello sviluppo intellettivo a basso livello adattativo [Capitolo/Saggio]
Bertelli, Marco O.; Vannucchi, Giulia; Persico, Antonio M.
abstract

Il presente capitolo insisterà sul trattamento psicofarmacologico della disabilità intellettiva o disturbo dello sviluppo intellettivo (DSI), secondo la terminologia più recente a basso livello adattativo (DSI-BA) e del disturbo dello spettro autistico (DSA) a basso funzionamento (DSA-BF), ovvero sulla frazione dei disturbi del neurosviluppo che mostra il maggior grado di disabilità, nell'ambito della comunicazione, della concettualizzazione, delle relazioni interpersonali e delle comuni attività della vita. Gli interventi psicofarmacologici rivolti a queste persone risultano fortemente caratterizzati, soprattutto rispetto ai criteri di prescrizione, alla tollerabilità e all'efficacia. Invece la distinzione delle specificità terapeutiche per il DSI-BA e per il DSA-BF è spesso difficile, sia in riferimento alla letteratura che all'esperienza clinica. Ciò è in parte dovuto alla sovrapposizione della fenomenica clinica: gli studi epidemiologici indicano tassi di comorbidità molto elevati, soprattutto nelle forme con QI basso. Difficoltà cognitive, d'apprendimento e d'adattamento precoci vengono riportate nel 25-70% delle persone con diagnosi di DSA e viceversa aspetti autistici sono stati rilevati nel 70% delle persone con DSI. Nei bambini con DSA la compresenza di DSI sembra determinare esiti peggiori in età adulta, soprattutto per quanto concerne la comunicazione verbale, i comportamenti ripetitivi e gli interessi ristretti. Una più grave compromissione cognitiva è risultata inoltre associata a una peggiore qualità delle interazioni sociali, della comunicazione reciproca e a maggiori deficit nel linguaggio espressivo e recettivo. Negli adolescenti e negli adulti con DSI la compresenza di DSA aumenta il numero e la gravità di comportamenti problematici e ha un impatto negativo sullo sviluppo di abilità sociali superiore a quello della psicosi.

2018 - Unraveling molecular pathways shared by Kabuki and Kabuki-like syndromes [Articolo su rivista]
Lintas, C.; Persico, A. M.
abstract

Kabuki syndrome (KS) is a rare genetic syndrome characterized by a typical facial gestalt, variable degrees of intellectual disability, organ malformations, postnatal growth retardation and skeletal abnormalities. So far, KMT2D or KDM6A mutation has been identified as the main cause of KS, accounting for 56%-75% and 3%-8% of cases, respectively. Patients without mutations in 1 of the 2 causative KS genes are often referred to as affected by Kabuki-like syndrome. Overall, they represent approximately 30% of KS cases, pointing toward substantial genetic heterogeneity for this condition. Here, we review all currently available literature describing KS-like phenotypes (or phenocopies) associated with genetic variants located in loci different from KMT2D and KDM6A . We also report on a new KS phenocopy harboring a 5 Mb de novo deletion in chr10p11.22-11.21. An enrichment analysis aimed at identifying functional Gene Ontology classes shared by the 2 known KS causative genes and by new candidate genes currently associated with KS-like phenotypes primarily converges upon abnormal chromatin remodeling and transcriptional dysregulation as pivotal to the pathophysiology of KS phenotypic hallmarks. The identification of mutations in genes belonging to the same functional pathways of KMT2D and KDM6A can help design molecular screenings targeted to KS-like phenotypes.

2017 - Copy number variation in 19 Italian multiplex families with autism spectrum disorder: Importance of synaptic and neurite elongation genes [Articolo su rivista]
Lintas, Carla; Picinelli, Chiara; Piras, Ignazio Stefano; Sacco, Roberto; Brogna, Claudia; Persico, Antonio M.
abstract

Autism Spectrum Disorder (ASD) is endowed with impressive heritability estimates and high recurrence rates. Its genetic underpinnings are nonetheless very heterogeneous, with common, and rare contributing variants located in hundreds of different loci, each characterized by variable levels of penetrance. Multiplex families from single ethnic groups represent a useful means to reduce heterogeneity and enhance genetic load. We screened 19 Italian ASD multiplex families (3 triplets and 16 duplets, total N = 41 ASD subjects), using array-CGH (Agilent 180 K). Causal or ASD-relevant CNVs were detected in 36.6% (15/41) of ASD probands, corresponding to 36.8% (7/19) multiplex families with at least one affected sibling genetically positive. However, only in less than half (3/7) of positive families, affected siblings share the same causal or ASD-relevant CNV. Even in these three families, additional potentially relevant CNVs not shared by affected sib pairs were also detected. These results provide further evidence of genetic heterogeneity in ASD even within multiplex families belonging to a single ethnic group. Differences in CNV burden may likely contribute to the substantial clinical heterogeneity observed between affected siblings. In addition, Gene Ontology enrichment analysis indicates that most potentially causal or relevant ASD genes detected in our cohort belong to nervous system-specific categories, especially involved in neurite elongation and synaptic structure/function. These findings point toward the existence of genomic instability in these families, whose underlying genetic and epigenetic mechanisms deserve further scrutiny.

2017 - Rates, distribution and implications of postzygotic mosaic mutations in autism spectrum disorder [Articolo su rivista]
Lim, Elaine T.; Uddin, Mohammed; De Rubeis, Silvia; Chan, Yingleong; Kamumbu, Anne S.; Zhang, Xiaochang; D'Gama, Alissa M.; Kim, Sonia N.; Hill, Robert Sean; Goldberg, Arthur P.; Poultney, Christopher; Minshew, Nancy J.; Kushima, Itaru; Aleksic, Branko; Ozaki, Norio; Parellada, Mara; Arango, Celso; Penzol, Maria J.; Carracedo, Angel; Kolevzon, Alexander; Hultman, Christina M.; Weiss, Lauren A.; Fromer, Menachem; Chiocchetti, Andreas G.; Freitag, Christine M.; Church, George M.; Scherer, Stephen W.; Buxbaum, Joseph D.; Walsh, Christopher A; Aleksic, B; Anney, R; Barbosa, M; Barrett, J; Betancur, C; Bishop, S; Brusco, A; Buxbaum, Jd; Carracedo, A; Chiocchetti, Ag; Chung, Bhy; Cook, E; Coon, H; Cutler, Dj; Daly, M; De Rubeis, S; Doan, R; Fernández-Prieto, M; Ferrero, Gb; Freitag, Cm; Fromer, M; Gargus, J; Geschwind, D; Gill, M; Gómez-Guerrero, L; Hansen-Kiss, E; He, X; Herman, G; Hertz-Picciotto, I; Hultman, C; Iliadou, B; Ionita-Laza, I; Jugessur, A; Knudsen, Gp; Kolevzon, A; Kosmicki, J; Kushima, I; Lee, Sl; Lehner, T; Lennertz, S; Lim, E; Maciel, P; Magnus, P; Manoach, D; Minshew, N; Morrow, E; Mulle, J; Neale, B; Ozaki, N; Palotie, A; Parellada, M; Passos-Bueno, Mr; Pericak-Vance, M; Persico, A; Pessah, I; Reichenberg, A; Reichert, J; Renieri, A; Robinson, E; Samocha, K; Sanders, S; Sandin, S; Santangelo, Sl; Satterstrom, K; Schafer, C; Schellenberg, G; Scherer, S; Senthil, G; Silva, M; Singh, T; Siper, Pm; Soares, G; Stevens, C; Stoltenberg, C; Surén, P; Sutcliffe, Js; Szatmari, P; Tassone, F; Thurm, A; Walsh, C; Weiss, L; Werling, D; Willsey, J; Xu, X; Yu, Tw; Yuen, R; Zwick, Me.
abstract

We systematically analyzed postzygotic mutations (PZMs) in whole-exome sequences from the largest collection of trios (5,947) with autism spectrum disorder (ASD) available, including 282 unpublished trios, and performed resequencing using multiple independent technologies. We identified 7.5% of de novo mutations as PZMs, 83.3% of which were not described in previous studies. Damaging, nonsynonymous PZMs within critical exons of prenatally expressed genes were more common in ASD probands than controls (P < 1 × 10-6), and genes carrying these PZMs were enriched for expression in the amygdala (P = 5.4 × 10-3). Two genes (KLF16 and MSANTD2) were significantly enriched for PZMs genome-wide, and other PZMs involved genes (SCN2A, HNRNPU and SMARCA4) whose mutation is known to cause ASD or other neurodevelopmental disorders. PZMs constitute a significant proportion of de novo mutations and contribute importantly to ASD risk.

2017 - The EU-AIMS Longitudinal European Autism Project (LEAP): clinical characterisation [Articolo su rivista]
Charman, Tony; Loth, Eva; Tillmann, Julian; Crawley, Daisy; Wooldridge, Caroline; Goyard, David; Ahmad, Jumana; Auyeung, Bonnie; Ambrosino, Sara; Banaschewski, Tobias; Baron-Cohen, Simon; Baumeister, Sarah; Beckmann, Christian; Bölte, Sven; Bourgeron, Thomas; Bours, Carsten; Brammer, Michael; Brandeis, Daniel; Brogna, Claudia; De Bruijn, Yvette; Chakrabarti, Bhismadev; Cornelissen, Ineke; Acqua, Flavio Dell'; Dumas, Guillaume; Durston, Sarah; Ecker, Christine; Faulkner, Jessica; Frouin, Vincent; Garcés, Pilar; Ham, Lindsay; Hayward, Hannah; Hipp, Joerg; Holt, Rosemary J.; Isaksson, Johan; Johnson, Mark H.; Jones, Emily J. H.; Kundu, Prantik; Lai, Meng-Chuan; D'Ardhuy, Xavier Liogier; Lombardo, Michael V.; Lythgoe, David J; Mandl, René; Mason, Luke; Meyer-Lindenberg, Andreas; Moessnang, Carolin; Mueller, Nico; O'Dwyer, Laurence; Oldehinkel, Marianne; Oranje, Bob; Pandina, Gahan; Persico, Antonio M.; Ruggeri, Barbara; Ruigrok, Amber N. V.; Sabet, Jessica; Sacco, Roberto; Cáceres, Antonia San Jóse; Simonoff, Emily; Toro, Roberto; Tost, Heike; Waldman, Jack; Williams, Steve C. R.; Zwiers, Marcel P.; Spooren, Will; Murphy, Declan G. M.; Buitelaar, Jan K.
abstract

BACKGROUND: The EU-AIMS Longitudinal European Autism Project (LEAP) is to date the largest multi-centre, multi-disciplinary observational study on biomarkers for autism spectrum disorder (ASD). The current paper describes the clinical characteristics of the LEAP cohort and examines age, sex and IQ differences in ASD core symptoms and common co-occurring psychiatric symptoms. A companion paper describes the overall design and experimental protocol and outlines the strategy to identify stratification biomarkers. METHODS: From six research centres in four European countries, we recruited 437 children and adults with ASD and 300 controls between the ages of 6 and 30 years with IQs varying between 50 and 148. We conducted in-depth clinical characterisation including a wide range of observational, interview and questionnaire measures of the ASD phenotype, as well as co-occurring psychiatric symptoms. RESULTS: The cohort showed heterogeneity in ASD symptom presentation, with only minimal to moderate site differences on core clinical and cognitive measures. On both parent-report interview and questionnaire measures, ASD symptom severity was lower in adults compared to children and adolescents. The precise pattern of differences varied across measures, but there was some evidence of both lower social symptoms and lower repetitive behaviour severity in adults. Males had higher ASD symptom scores than females on clinician-rated and parent interview diagnostic measures but not on parent-reported dimensional measures of ASD symptoms. In contrast, self-reported ASD symptom severity was higher in adults compared to adolescents, and in adult females compared to males. Higher scores on ASD symptom measures were moderately associated with lower IQ. Both inattentive and hyperactive/impulsive ADHD symptoms were lower in adults than in children and adolescents, and males with ASD had higher levels of inattentive and hyperactive/impulsive ADHD symptoms than females. CONCLUSIONS: The established phenotypic heterogeneity in ASD is well captured in the LEAP cohort. Variation both in core ASD symptom severity and in commonly co-occurring psychiatric symptoms were systematically associated with sex, age and IQ. The pattern of ASD symptom differences with age and sex also varied by whether these were clinician ratings or parent- or self-reported which has important implications for establishing stratification biomarkers and for their potential use as outcome measures in clinical trials.

2017 - The EU-AIMS Longitudinal European Autism Project (LEAP): design and methodologies to identify and validate stratification biomarkers for autism spectrum disorders [Articolo su rivista]
Loth, Eva; Charman, Tony; Mason, Luke; Tillmann, Julian; Jones, Emily J. H.; Wooldridge, Caroline; Ahmad, Jumana; Auyeung, Bonnie; Brogna, Claudia; Ambrosino, Sara; Banaschewski, Tobias; Baron-Cohen, Simon; Baumeister, Sarah; Beckmann, Christian; Brammer, Michael; Brandeis, Daniel; Bölte, Sven; Bourgeron, Thomas; Bours, Carsten; De Bruijn, Yvette; Chakrabarti, Bhismadev; Crawley, Daisy; Cornelissen, Ineke; Acqua, Flavio Dell'; Dumas, Guillaume; Durston, Sarah; Ecker, Christine; Faulkner, Jessica; Frouin, Vincent; Garces, Pilar; Goyard, David; Hayward, Hannah; Ham, Lindsay M.; Hipp, Joerg; Holt, Rosemary J.; Johnson, Mark H.; Isaksson, Johan; Kundu, Prantik; Lai, Meng-Chuan; D'Ardhuy, Xavier Liogier; Lombardo, Michael V.; Lythgoe, David J.; Mandl, René; Meyer-Lindenberg, Andreas; Moessnang, Carolin; Mueller, Nico; O'Dwyer, Laurence; Oldehinkel, Marianne; Oranje, Bob; Pandina, Gahan; Persico, Antonio M.; Ruigrok, Amber N. V.; Ruggeri, Barbara; Sabet, Jessica; Sacco, Roberto; Cáceres, Antonia San José; Simonoff, Emily; Toro, Roberto; Tost, Heike; Waldman, Jack; Williams, Steve C. R.; Zwiers, Marcel P.; Spooren, Will; Murphy, Declan G. M.; Buitelaar, Jan K.
abstract

BACKGROUND: The tremendous clinical and aetiological diversity among individuals with autism spectrum disorder (ASD) has been a major obstacle to the development of new treatments, as many may only be effective in particular subgroups. Precision medicine approaches aim to overcome this challenge by combining pathophysiologically based treatments with stratification biomarkers that predict which treatment may be most beneficial for particular individuals. However, so far, we have no single validated stratification biomarker for ASD. This may be due to the fact that most research studies primarily have focused on the identification of mean case-control differences, rather than within-group variability, and included small samples that were underpowered for stratification approaches. The EU-AIMS Longitudinal European Autism Project (LEAP) is to date the largest multi-centre, multi-disciplinary observational study worldwide that aims to identify and validate stratification biomarkers for ASD. METHODS: LEAP includes 437 children and adults with ASD and 300 individuals with typical development or mild intellectual disability. Using an accelerated longitudinal design, each participant is comprehensively characterised in terms of clinical symptoms, comorbidities, functional outcomes, neurocognitive profile, brain structure and function, biochemical markers and genomics. In addition, 51 twin-pairs (of which 36 had one sibling with ASD) are included to identify genetic and environmental factors in phenotypic variability. RESULTS: Here, we describe the demographic characteristics of the cohort, planned analytic stratification approaches, criteria and steps to validate candidate stratification markers, pre-registration procedures to increase transparency, standardisation and data robustness across all analyses, and share some 'lessons learnt'. A clinical characterisation of the cohort is given in the companion paper (Charman et al., accepted). CONCLUSION: We expect that LEAP will enable us to confirm, reject and refine current hypotheses of neurocognitive/neurobiological abnormalities, identify biologically and clinically meaningful ASD subgroups, and help us map phenotypic heterogeneity to different aetiologies.

2016 - Difference in Visual Social Predispositions between Newborns at Low-and High-risk for Autism [Articolo su rivista]
Di Giorgio, Elisa; Frasnelli, Elisa; Rosa Salva, Orsola; Maria Luisa, Scattoni; Puopolo, Maria; Tosoni, Daniela; Simion, Francesca; Vallortigara, Giorgio; Apicella, Fabio; Gagliano, Antonella; Guzzetta, Andrea; Molteni, Massimo; Persico, Antonio; Pioggia, Giovanni; Valeri, Giovanni; Vicari, Stefano
abstract

2016 - Differential methylation at the RELN gene promoter in temporal cortex from autistic and typically developing post-puberal subjects [Articolo su rivista]
Lintas, Carla; Sacco, Roberto; Persico, Antonio
abstract

Background: Reelin plays a pivotal role in neurodevelopment and in postnatal synaptic plasticity, and has been implicated in the pathogenesis of Autism Spectrum Disorder (ASD). RELN gene expression is significantly decreased in ASD, both in the brain and peripherally. Methylation at the RELN gene promoter is largely triggered at puberty and hypermethylation has been found in post-mortem brains of schizophrenic and bipolar patients. Methods: In this study, we assessed RELN gene methylation status in post-mortem temporocortical tissue samples (BA41/42 or 22) of six pairs of post-puberal individuals with ASD and typically developing subjects, matched for sex (M:F=5:1), age and post-mortem interval. Results: ASD patients display a significantly higher number of methylated CpG islands and heavier methylation in the 5’ region of the RELN gene promoter, spanning from -458 to -223 bp, whereas controls have more methylated CpG positions and greater extent of methylation at the 3’ promoter region, spanning from -222 to +1 bp. The most upstream promoter region (-458 to -364 bp) is methylated only in ASD brains, while the most downstream region (-131 to +1 bp) is methylated exclusively in control brains. Within this general framework, three different methylation patterns are discernible, each correlated with different extents of reduction in reelin gene expression among ASD individuals compared to controls. Conclusions: The methylation pattern is different in ASD and control post-mortem brains. ASD-specific CpG positions, located in the most upstream gene promoter region, may exert a functional role potentially conferring ASD risk by blunting RELN gene expression.

2016 - Identification and validation of biomarkers for autism spectrum disorders [Articolo su rivista]
Loth, E.; Spooren, W.; Ham, L. M.; Isaac, M. B.; Auriche-Benichou, C.; Banaschewski, T.; Baron-Cohen, S.; Broich, K.; Bolte, S.; Bourgeron, T.; Charman, T.; Collier, D.; de Andres-Trelles, F.; Durston, S.; Ecker, C.; Elferink, A.; Haberkamp, M.; Hemmings, R.; Johnson, M. H.; Jones, E. J. H.; Khwaja, O. S.; Lenton, S.; Mason, L.; Mantua, V.; Meyer-Lindenberg, A.; Lombardo, M. V.; O'Dwyer, L.; Okamoto, K.; Pandina, G. J.; Pani, L.; Persico, A. M.; Simonoff, E.; Tauscher-Wisniewski, S.; Llinares-Garcia, J.; Vamvakas, S.; Williams, S.; Buitelaar, J. K.; Murphy, D. G. M.
abstract

2016 - Recurrent 15q11.2 BP1-BP2 microdeletions and microduplications in the etiology of neurodevelopmental disorders [Articolo su rivista]
Picinelli, Chiara; Lintas, Carla; Piras, Ignazio Stefano; Gabriele, Stefano; Sacco, Roberto; Brogna, Claudia; Persico, Antonio
abstract

Rare and common CNVs can contribute to the etiology of neurodevelopmental disorders. One of the recurrent genomic aberrations associated with these phenotypes and proposed as a susceptibility locus is the 15q11.2 BP1-BP2 CNV encompassing TUBGCP5, CYFIP1, NIPA2 and NIPA1. Characterizing by array-CGH a cohort of 243 families with various neurodevelopmental disorders, we identified 5 patients carrying the 15q11.2 duplication and one carrying the deletion. All CNVs were confirmed by qPCR and were inherited, except for one duplication where parents were not available. The phenotypic spectrum of CNV carriers was broad but mainly neurodevelopmental, in line with all four genes being implicated in axonal growth and neural connectivity. Phenotypically normal and mildly affected carriers complicate the interpretation of this aberration. This variability may be due to reduced penetrance or altered gene dosage on a particular genetic background. We evaluated the expression levels of the four genes in peripheral blood RNA and found the expected reduction in the deleted case, while duplicated carriers displayed high interindividual variability. These data suggest that differential expression of these genes could partially account for differences in clinical phenotypes, especially among duplication carriers. Furthermore, urinary Mg2+ levels appear negatively correlated with NIPA2 gene copy number, suggesting they could potentially represent a useful biomarker, whose reliability will need replication in larger samples.

2016 - Slow intestinal transit contributes to elevate urinary p-cresol level in Italian autistic children [Articolo su rivista]
Gabriele, Stefano; Sacco, Roberto; Altieri, Laura; Neri, Cristina; Urbani, Andrea; Bravaccio, Carmela; Riccio, Maria Pia; Iovene, Maria Rosaria; Bombace, Francesca; De Magistris, Laura; Persico, Antonio
abstract

The uremic toxin p-cresol (4-methylphenol) is either of environmental origin or can be synthetized from tyrosine by cresol-producing bacteria present in the gut lumen. Elevated p-cresol amounts have been previously found in the urines of Italian and French autism spectrum disorder (ASD) children up until 8 years of age, and may be associated with autism severity or with the intensity of abnormal behaviors. This study aims to investigate the mechanism producing elevated urinary p-cresol in ASD. Urinary p-cresol levels were thus measured by High Performance Liquid Chromatography in a sample of 53 Italian ASD children assessed for (a) presence of Clostridium spp. strains in the gut by means of an in vitro fecal stool test and of Clostridium difficile-derived toxin A/B in the feces, (b) intestinal permeability using the lactulose/mannitol (LA/MA) test, (c) frequent use of antibiotics due to recurrent infections during the first 2 years of postnatal life, and (d) stool habits with the Bristol Stool Form Scale. Chronic constipation was the only variable significantly associated with total urinary p-cresol concentration (P < 0.05). No association was found with presence of Clostridium spp. in the gut flora (P50.92), augmented intestinal permeability (P50.18), or frequent use of antibiotics in early infancy (P50.47). No ASD child was found to carry C. difficile in the gut or to release toxin A/B in the feces. In conclusion, urinary p-cresol levels are elevated in young ASD children with increased intestinal transit time and chronic constipation.

2016 - Urinary metabolomics of young Italian autistic children supports abnormal tryptophan and purine metabolism [Articolo su rivista]
Gevi, Federica; Zolla, Lello; Gabriele, Stefano; Persico, Antonio M.
abstract

BACKGROUND: Autism spectrum disorder (ASD) is still diagnosed through behavioral observation, due to a lack of laboratory biomarkers, which could greatly aid clinicians in providing earlier and more reliable diagnoses. Metabolomics on human biofluids provides a sensitive tool to identify metabolite profiles potentially usable as biomarkers for ASD. Initial metabolomic studies, analyzing urines and plasma of ASD and control individuals, suggested that autistic patients may share some metabolic abnormalities, despite several inconsistencies stemming from differences in technology, ethnicity, age range, and definition of "control" status. METHODS: ASD-specific urinary metabolomic patterns were explored at an early age in 30 ASD children and 30 matched controls (age range 2-7, M:F = 22:8) using hydrophilic interaction chromatography (HILIC)-UHPLC and mass spectrometry, a highly sensitive, accurate, and unbiased approach. Metabolites were then subjected to multivariate statistical analysis and grouped by metabolic pathway. RESULTS: Urinary metabolites displaying the largest differences between young ASD and control children belonged to the tryptophan and purine metabolic pathways. Also, vitamin B6, riboflavin, phenylalanine-tyrosine-tryptophan biosynthesis, pantothenate and CoA, and pyrimidine metabolism differed significantly. ASD children preferentially transform tryptophan into xanthurenic acid and quinolinic acid (two catabolites of the kynurenine pathway), at the expense of kynurenic acid and especially of melatonin. Also, the gut microbiome contributes to altered tryptophan metabolism, yielding increased levels of indolyl 3-acetic acid and indolyl lactate. CONCLUSIONS: The metabolic pathways most distinctive of young Italian autistic children largely overlap with those found in rodent models of ASD following maternal immune activation or genetic manipulations. These results are consistent with the proposal of a purine-driven cell danger response, accompanied by overproduction of epileptogenic and excitotoxic quinolinic acid, large reductions in melatonin synthesis, and gut dysbiosis. These metabolic abnormalities could underlie several comorbidities frequently associated to ASD, such as seizures, sleep disorders, and gastrointestinal symptoms, and could contribute to autism severity. Their diagnostic sensitivity, disease-specificity, and interethnic variability will merit further investigation.

2016 - Xp22.33p22.12 Duplication in a Patient with Intellectual Disability and Dysmorphic Facial Features [Articolo su rivista]
Lintas, Carla; Picinelli, Chiara; Piras, Ignazio S.; Sacco, Roberto; Gabriele, Stefano; Verdecchia, Magda; Persico, Antonio
abstract

A novel 19.98 Mb duplication on chromosome Xp22.33-p22.12 was detected by array CGH in a 30 year old man affected by intellectual disability, congenital hypotonia and dysmorphic traits. The duplication encompasses more than 113 known genes. Many of these genes (as neuroligin 4, cyclin-dependent kinase like 5 and others) have already correlated with X-linked intellectual disability and/or neurodevelopmental disorders. Due to the high number of potentially pathogenic genes involved in the reported duplication, we cannot correlate the clinical phenotype to a single gene. Indeed, we suggest that the resulting clinical phenotype may have arisen from the overexpression and consequent perturbation of fine gene dosage.

2015 - Autism genetics: Methodological issues and experimental design [Articolo su rivista]
Sacco, Roberto; Lintas, Carla; Persico, Antonio
abstract

Autism is a complex neuropsychiatric disorder of developmental origin, where multiple genetic and environmental factors likely interact resulting in a clinical continuum between “affected” and “unaffected” individuals in the general population. During the last two decades, relevant progress has been made in identifying chromosomal regions and genes in linkage or association with autism, but no single gene has emerged as a major cause of disease in a large number of patients. The purpose of this paper is to discuss specific methodological issues and experimental strategies in autism genetic research, based on fourteen years of experience in patient recruitment and association studies of autism spectrum disorder in Italy.

2015 - Endocannabinoid Signaling in Autism [Articolo su rivista]
Chakrabarti, B.; Persico, A.; Battista, N.; Maccarrone, M.
abstract

Autism spectrum disorder (ASD) is a complex behavioral condition with onset during early childhood and a lifelong course in the vast majority of cases. To date, no behavioral, genetic, brain imaging, or electrophysiological test can specifically validate a clinical diagnosis of ASD. However, these medical procedures are often implemented in order to screen for syndromic forms of the disorder (i.e., autism comorbid with known medical conditions). In the last 25&nbsp;years a good deal of information has been accumulated on the main components of the “endocannabinoid (eCB) system”, a rather complex ensemble of lipid signals (“endocannabinoids”), their target receptors, purported transporters, and metabolic enzymes. It has been clearly documented that eCB signaling plays a key role in many human health and disease conditions of the central nervous system, thus opening the avenue to the therapeutic exploitation of eCB-oriented drugs for the treatment of psychiatric, neurodegenerative, and neuroinflammatory disorders. Here we present a modern view of the eCB system, and alterations of its main components in human patients and animal models relevant to ASD. This review will thus provide a critical perspective necessary to explore the potential exploitation of distinct elements of eCB system as targets of innovative therapeutics against ASD.

2015 - Environmental factors and Autism Spectrum Disorder [Capitolo/Saggio]
Persico, A. M.; Merelli, S.
abstract

Multiple observations indicate that environmental and epigenetic factors play an important role in the emergence of Autism Spectrum Disorder (ASD). Growing ASD incidence rates, the incomplete penetrance of many rare variants linked to autism, and the increased exposure to environmental contaminants, all strongly support the role of Gene x Environment interactions in a substantial fraction of autistic patients. Within this framework, genetically susceptible individuals exposed to detrimental environmental factors at critical times during neurodevelopment, might undergo disrupted brain morphogenesis, neuronal connectivity, and synaptic functioning consequently yielding ASD. Several teratogenic drugs and prenatal viral infections are able to cause autism in humans, as supported by case reports, cohort studies and animal models. Moreover recent studies are showing that some newly identified potential neurotoxicants may negatively affect developmental trajectories, leading to altered cognitive, attentive, behavioral and motor performance, as well as to systemic abnormalities frequently seen in autistic individuals. A variety of mechanisms is potentially involved, ranging from oxidative and inflammatory brain damage to altered gene expression and impaired signal transduction. More research is needed to thoroughly investigate the effects of these compounds on neurodevelopment, to validate their involvement specifically in ASD, to study Gene x Environment interactions in potentially susceptible individuals and to plan targeted prevention strategies.

2015 - Genetica e terapia: verso un trattamento individualizzato [Capitolo/Saggio]
Persico, A. M.
abstract

2015 - Head circumference and brain size in autism spectrum disorder: A systematic review and meta-analysis [Articolo su rivista]
Sacco, Roberto; Gabriele, Stefano; Persico, Antonio
abstract

Macrocephaly and brain overgrowth have been associated with autism spectrum disorder. We performed a systematic review and meta-analysis to provide an overall estimate of effect size and statistical significance for both head circumference and total brain volume in autism. Our literature search strategy identified 261 and 391 records, respectively; 27 studies defining percentages of macrocephalic patients and 44 structural brain imaging studies providing total brain volumes for patients and controls were included in our meta-analyses. Head circumference was significantly larger in autistic compared to control individuals, with 822/5225 (15.7%) autistic individuals displaying macrocephaly. Structural brain imaging studies measuring brain volume estimated effect size. The effect size is higher in low functioning autistics compared to high functioning and ASD individuals. Brain overgrowth was recorded in 142/1558 (9.1%) autistic patients. Finally, we found a significant interaction between age and total brain volume, resulting in larger head circumference and brain size during early childhood. Our results provide conclusive effect sizes and prevalence rates for macrocephaly and brain overgrowth in autism, confirm the variation of abnormal brain growth with age, and support the inclusion of this endophenotype in multi-biomarker diagnostic panels for clinical use.

2015 - Migraine genetics: current findings and future lines of research [Articolo su rivista]
Persico, A. M.; Verdecchia, M.; Pinzone, V.; Guidetti, V.
abstract

In the last two decades, migraine research has greatly advanced our current knowledge of the genetic contributions and the pathophysiology of this common and debilitating disorder. Nonetheless, this knowledge still needs to grow further and to translate into more effective treatments. To date, several genes involved in syndromic and monogenic forms of migraine have been identified, allowing the generation of animal models which have significantly contributed to current knowledge of the mechanisms underlying these rare forms of migraine. Common forms of migraine are instead posing a greater challenge, as they may most often stem from complex interactions between multiple common genetic variants, with environmental triggers. This paper reviews our current understanding of migraine genetics, moving from syndromic and monogenic forms to oligogenic/polygenic migraines most recently addressed with some success through genome-wide association studies. Methodological issues in study design and future perspectives opened by biomarker research will also be briefly addressed.

2015 - Preliminary Transcriptome Analysis in Lymphoblasts from Cluster Headache and Bipolar Disorder Patients Implicates Dysregulation of Circadian and Serotonergic Genes [Articolo su rivista]
Costa, Marta; Squassina, Alessio; Piras, Ignazio Stefano; Pisanu, Claudia; Congiu, Donatella; Niola, Paola; Angius, Andrea; Chillotti, Caterina; Ardau, Raffaella; Severino, Giovanni; Stochino, Erminia; Deidda, Arianna; Persico, Antonio; Alda, Martin; Del Zompo, Maria
abstract

Bipolar disorder (BD) and cluster headache (CH) are distinct conditions with important similarities such as a temporal pattern of disturbances, dysregulation of the sleep–wake cycle, and response to lithium treatment in a proportion of patients. Aiming to identify common transcription signatures in these two disorders, we carried out an exploratory microarray gene expression analysis in lymphoblasts from 8 CH and 10 BD I patients selected for positive response to lithium and 10 healthy controls (CO). Gene expression levels of BD and CH were compared with CO to create two lists of differentially expressed genes. We then matched the two lists and focus on genes showing statistically significant difference and same change direction in both disorders. RNA binding motif protein 3 (RBM3) was the most significantly altered gene in the list (3.17 × 10−13 in BD, 9.44 × 10−14 in CH). Pathway analysis identified protein processing in endoplasmic reticulum as the most significantly enriched. For validation with quantitative reverse transcription PCR (qRT-PCR) using the same samples, we selected seven genes. Among these, we were able to validate the RBM3, nuclear receptor subfamily 1, group D, member 1 (NR1D1), and tryptophan hydroxylase 1 (TPH1). These genes encode for elements involved in circadian rhythm regulation (RBM3 and NR1D1) and in serotonin synthesis (TPH1), processes previously involved in both disorders, and in the mechanism of action of lithium.

2015 - Unmet needs in paediatric psychopharmacology: Present scenario and future perspectives [Articolo su rivista]
Persico, Antonio; Arango, Celso; Buitelaar, Jan K.; Correll, Christoph U.; Glennon, Jeffrey C.; Hoekstra, Pieter J.; Moreno, Carmen; Vitiello, Benedetto; Vorstman, Jacob; Zuddas, Alessandro; Banaschewski, Tobias; Castro-Fornieles, Josefina; Coghill, David; Cohen, David; Dittmann, Ralf W.; Fegert, Jörg M.; Purper Ouakil, Diane; Parellada, Mara; Roessner, Veit
abstract

Paediatric psychopharmacology holds great promise in two equally important areas of enormous biomedical and social impact, namely the treatment of behavioural abnormalities in children and adolescents, and the prevention of psychiatric disorders with adolescent- or adult-onset. Yet, in striking contrast, pharmacological treatment options presently available in child and adolescent psychiatry are dramatically limited. The most important currently unmet needs in paediatric psychopharmacology are: the frequent off-label prescription of medications to children and adolescents based exclusively on data from randomized controlled studies involving adult patients; the frequent lack of age-specific dose, long-term efficacy and tolerability/safety data; the lack of effective medications for many paediatric psychiatric disorders, most critically autism spectrum disorder; the scarcity and limitations of randomized placebo-controlled trials in paediatric psychopharmacology; the unexplored potential for the prevention of psychiatric disorders with adolescent- and adult-onset; the current lack of biomarkers to predict treatment response and severe adverse effects; the need for better preclinical data to foster the successful development of novel drug therapies; and the effective dissemination of evidence-based treatments to the general public, to better inform patients and families of the benefits and risks of pharmacological interventions during development. Priorities and strategies are proposed to overcome some of these limitations, including the European Child and Adolescent Clinical Psychopharmacology Network, as an overarching Pan-European infrastructure aimed at reliably carrying out much needed psychopharmacological trials in children and adolescents, in order to fill the identified gaps and improve overall outcomes.

2014 - Anti-brain antibodies are associated with more severe cognitive and behavioral profiles in Italian children with Autism Spectrum Disorder [Articolo su rivista]
Piras, I S; Haapanen, L; Napolioni, V; Sacco, R; Van de Water, J; Persico, A M
abstract

Circulating 45 and 62kDa antibodies targeting the cerebellum were previously associated with Autism Spectrum Disorder (ASD), lower adaptive/cognitive function and aberrant behaviors. Moreover, 37, 39 and 73kDa maternal antibodies (mAb) targeting the fetal brain were previously correlated with broad autism spectrum, irritability, abnormal brain enlargement and impaired expressive language. The present study aims towards clinically characterizing individuals with brain-targeted IgG and/or exposed to maternal antibrain antibodies in a large sample of Italian autistic children (N=355), their unaffected siblings (N=142) and mothers (N=333). The presence of patient- and mother-produced anti-brain antibodies does not confer increased risk of autism within the same sibship. However, the 45 and 62kDa antibodies are correlated with autism severity: the 45kDa Ab is associated with cognitive impairment and lower scores at the Vineland Adaptive Behavior Scales, the 62kDa Ab with motor stereotypies, while both correlate with larger head circumference (all P<0.05). On the other hand, maternal 37, 39 and 73kDa antibrain antibodies, either alone or in combination, are correlated with impaired verbal and non-verbal language development, neurodevelopmental delay and sleep/wake cycle disturbances in their autistic children (P<0.05). Presence of the 62kDa autoAb in the child is significantly associated with presence of the 39 and/or 73kDa antibodies in his/her mother. Our results confirm and extend previous observations in an ethnically distinct sample, providing further evidence of a pathomorphic role for anti-brain antibodies in autism while demonstrating their familial clustering.

2014 - Biomarkers in autism spectrum disorder: The old and the new [Articolo su rivista]
Ruggeri, Barbara; Sarkans, Ugis; Schumann, Gunter; Persico, Antonio M.
abstract

RATIONALE: Autism spectrum disorder (ASD) is a complex heterogeneous neurodevelopmental disorder with onset during early childhood and typically a life-long course. The majority of ASD cases stems from complex, 'multiple-hit', oligogenic/polygenic underpinnings involving several loci and possibly gene-environment interactions. These multiple layers of complexity spur interest into the identification of biomarkers able to define biologically homogeneous subgroups, predict autism risk prior to the onset of behavioural abnormalities, aid early diagnoses, predict the developmental trajectory of ASD children, predict response to treatment and identify children at risk for severe adverse reactions to psychoactive drugs. OBJECTIVES: The present paper reviews (a) similarities and differences between the concepts of 'biomarker' and 'endophenotype', (b) established biomarkers and endophenotypes in autism research (biochemical, morphological, hormonal, immunological, neurophysiological and neuroanatomical, neuropsychological, behavioural), (c) -omics approaches towards the discovery of novel biomarker panels for ASD, (d) bioresource infrastructures and (e) data management for biomarker research in autism. RESULTS: Known biomarkers, such as abnormal blood levels of serotonin, oxytocin, melatonin, immune cytokines and lymphocyte subtypes, multiple neuropsychological, electrophysiological and brain imaging parameters, will eventually merge with novel biomarkers identified using unbiased genomic, epigenomic, transcriptomic, proteomic and metabolomic methods, to generate multimarker panels. Bioresource infrastructures, data management and data analysis using artificial intelligence networks will be instrumental in supporting efforts to identify these biomarker panels. CONCLUSIONS: Biomarker research has great heuristic potential in targeting autism diagnosis and treatment.

2014 - Blood serotonin levels in autism spectrum disorder: A systematic review and meta-analysis [Articolo su rivista]
Gabriele, Stefano; Sacco, Roberto; Persico, Antonio M.
abstract

Elevated blood serotonin (5-HT) levels were the first biomarker identified in autism research. Many studies have contrasted blood 5-HT levels in autistic patients and controls, but different measurement protocols, technologies, and biomaterials have been used through the years. We performed a systematic review and meta-analysis to provide an overall estimate of effect size and between-study heterogeneity, while verifying whether and to what extent different methodological approaches influence the strength of this association. Our literature search strategy identified 551 papers, from which 22 studies providing patient and control blood 5-HT values were selected for meta-analysis. Significantly higher 5-HT levels in autistic patients compared to controls were recorded both in whole blood (WB) [O.R.=4.6; (3.1-5.2); P=1.0×10(-12]), and in platelet-rich plasma (PRP) [O.R.=2.6 (1.8-3.9); P=2.7×10(-7)]. Predictably, studies measuring 5-HT levels in platelet-poor plasma (PPP) yielded no significant group difference [O.R.=0.54 (0.2-2-0); P=0.36]. Altogether, elevated 5-HT blood levels were recorded in 28.3% in WB and 22.5% in PRP samples of autistic individuals, as reported in 15 and 4 studies, respectively. Studies employing HPLC vs fluorometric assays yield similar cumulative effect sizes, but the former display much lower variability. In summary, despite some limitations mainly due to small study sample sizes, our results significantly reinforce the reliability of elevated 5-HT blood levels as a biomarker in ASD, providing practical indications potentially useful for its inclusion in multi-marker diagnostic panels for clinical use.

2014 - Endophenotypes in Autism Spectrum Disorders [Capitolo/Saggio]
Persico, Antonio Maria; Sacco, Roberto
abstract

Autism Spectrum Disorder (ASD) is a complex disease with onset during early childhood and typically a life-long course. Genetic liability plays a prominent role in ASD, as siblings of affected individuals display on average 25-fold higher autism risk rates compared to the general population. However, genetic contributions fit in most cases with complex, “multiple-hit”, oligogenic/polygenic models encompassing several loci and also gene-environment interactions. Furthermore, ASD should be viewed as the extreme end of a set of continuous dimensions pertaining to the realms of human socialization, language development and behaviour, rather than as a “black or white” medical condition. This complexity spurs interest into the heuristic potential of “endophenotypes” in ASD, as originally applied by Gottesman and Shields (1973) to the realm of psychiatry. Endophenotypes can be defined as familial, heritable and quantitative traits associated with a complex disease. Occupying an intermediate position between genotype and behaviour, these traits offer the potential to bridge the gap between complex disease phenotypes, such as autism, and the underlying genetic mechanisms. The most reliable endophenotypes in ASD can be grouped into seven categories: biochemical, morphological, hormonal, immunological, neurophysiological/neuroanatomical, neuropsychological and behavioural. Their use holds promise to foster advances not only toward more reliable genotype-phenotype correlations in autism research, but also in dissecting clinical subgroups of ASD patients with relatively homogeneous pathophysiological underpinnings, aiding clinicians in early diagnosis once incorporated into broader biomarker panels, and predicting developmental trajectories and treatment response.

2014 - Environmental Factors in the Onset of Autism Spectrum Disorder [Articolo su rivista]
Persico, Antonio M.; Merelli, Sara
abstract

Autism Spectrum Disorder (ASD) is a heterogeneous clinical condition, whose prevalence has grown considerably during the last decade. Genetic factors are thought to underpin the disorder, but common genetic variants and epigenetic mechanisms have been increasingly called into question for the majority of ASD cases. Growing prenatal exposure to new environmental toxicants has been shown to potentially affect brain development, leading to altered cognitive, social, attentive, behavioral and motor performance. Both epidemiological evidence and mechanistic studies assessing oxidative stress, neuroinflammation, epigenetic alterations and impaired signal transduction, all observed following neurotoxicant exposure, indeed lend biological plausibility to Gene x Environment interactions, whereby environmental toxicants interacting additively or synergistically with genetic liability, can push prenatal neurodevelopmental processes over the threshold for postnatal ASD expression. Research on environmental contributions to ASD and on specific Gene x Environment interaction models ultimately aims at defining targeted preventive strategies.

2014 - Stratified medicine for mental disorders [Articolo su rivista]
Schumann, Gunter; Binder, Elisabeth B.; Holte, Arne; de Kloet, E. Ronald; Oedegaard, Ketil J.; Robbins, Trevor W.; Walker-Tilley, Tom R.; Bitter, Istvan; Brown, Verity J.; Buitelaar, Jan; Ciccocioppo, Roberto; Cools, Roshan; Escera, Carles; Fleischhacker, Wolfgang; Flor, Herta; Frith, Chris D.; Heinz, Andreas; Johnsen, Erik; Kirschbaum, Clemens; Klingberg, Torkel; Lesch, Klaus-Peter; Lewis, Shon; Maier, Wolfgang; Mann, Karl; Martinot, Jean-Luc; Meyer-Lindenberg, Andreas; Müller, Christian P.; Müller, Walter E.; Nutt, David J.; Persico, Antonio; Perugi, Giulio; Pessiglione, Mathias; Preuss, Ulrich W.; Roiser, Jonathan P.; Rossini, Paolo M.; Rybakowski, Janusz K.; Sandi, Carmen; Stephan, Klaas E.; Undurraga, Juan; Vieta, Eduard; van der Wee, Nic; Wykes, Til; Haro, Josep Maria; Wittchen, Hans Ulrich
abstract

There is recognition that biomedical research into the causes of mental disorders and their treatment needs to adopt new approaches to research. Novel biomedical techniques have advanced our understanding of how the brain develops and is shaped by behaviour and environment. This has led to the advent of stratified medicine, which translates advances in basic research by targeting aetiological mechanisms underlying mental disorder. The resulting increase in diagnostic precision and targeted treatments may provide a window of opportunity to address the large public health burden, and individual suffering associated with mental disorders. While mental health and mental disorders have significant representation in the "health, demographic change and wellbeing" challenge identified in Horizon 2020, the framework programme for research and innovation of the European Commission (2014-2020), and in national funding agencies, clear advice on a potential strategy for mental health research investment is needed. The development of such a strategy is supported by the EC-funded "Roadmap for Mental Health Research" (ROAMER) which will provide recommendations for a European mental health research strategy integrating the areas of biomedicine, psychology, public health well being, research integration and structuring, and stakeholder participation. Leading experts on biomedical research on mental disorders have provided an assessment of the state of the art in core psychopathological domains, including arousal and stress regulation, affect, cognition social processes, comorbidity and pharmacotherapy. They have identified major advances and promising methods and pointed out gaps to be addressed in order to achieve the promise of a stratified medicine for mental disorders.

2014 - The GLO1 C332 (Ala111) allele confers autism vulnerability: family-based genetic association and functional correlates [Articolo su rivista]
Gabriele, Stefano; Lombardi, Federica; Sacco, Roberto; Napolioni, Valerio; Altieri, Laura; Tirindelli, Maria Cristina; Gregorj, Chiara; Bravaccio, Carmela; Rousseau, Francis; Persico, Antonio M
abstract

Glyoxalase I (GLO1) is a homodimeric Zn(2+)-dependent isomerase involved in the detoxification of methylglyoxal and in limiting the formation of advanced glycation end-products (AGE). We previously found the rs4746 A332 (Glu111) allele of the GLO1 gene, which encodes for glyoxalase I, associated with "unaffected sibling" status in families with one or more children affected by Autism Spectrum Disorder (ASD). To identify and characterize this protective allele, we sequenced GLO1 exons and exon-intron junctions, detecting two additional SNPs (rs1049346, rs1130534) in linkage disequilibrium with rs4746. A family-based association study involving 385 simplex and 20 multiplex Italian families yielded a significant association with autism driven only by the rs4746 C332 (Ala111) allele itself (P < 0.05 and P < 0.001 under additive and dominant/recessive models, respectively). Glyoxalase enzymatic activity was significantly reduced both in leukocytes and in post-mortem temporocortical tissue (N = 38 and 13, respectively) of typically developing C332 allele carriers (P < 0.05 and <0.01), with no difference in Glo1 protein levels. Conversely, AGE amounts were significantly higher in the same C332 post-mortem brains (P = 0.001), with a strong negative correlation between glyoxalase activity and AGE levels (τ = -0.588, P < 0.01). Instead, 19 autistic brains show a dysregulation of the glyoxalase-AGE axis (τ = -0.209, P = 0.260), with significant blunting of glyoxalase activity and AGE amounts compared to controls (P < 0.05), and loss of rs4746 genotype effects. In summary, the GLO1 C332 (Ala111) allele confers autism vulnerability by reducing brain glyoxalase activity and enhancing AGE formation, but years after an autism diagnosis the glyoxalase-AGE axis appears profoundly disrupted, with loss of C332 allelic effects.

2014 - Urinary p-Cresol in ASD [Capitolo/Saggio]
Persico, Antonio Maria; Napolioni, Valerio
abstract

The etiology of autism encompasses a broad range of causative events, ranging from rare de novo high-penetrance mutations affecting genes such as NLGN3/4, SHANK3, NRXN1, and MECP2, to a complex mix of environmental and epigenetic factors acting upon a vulnerable genetic background. Increasing prevalence rates, decreasing heritability estimates, polygenic models with multiple incompletely penetrant de novo mutations, all suggest that, in addition to broader diagnostic criteria and increased awareness, also a real increase in incidence primarily due to greater gene-environment interactions may also be occurring. Notably, the phenotypic heterogeneity of ASD suggests the existence of many “autisms”, each characterized by specific etiopathogenetic underpinnings. Given the complex nature of this disorder, great effort is now underway aiming to define a reliable panel of biological markers able to assist clinicians in an early diagnosis. P-cresol (4-methylphenol) belongs to the cresol class of organic aromatic compounds. Environmental p-cresol is absorbed through the gastrointestinal and the respiratory tracts, as well as through the intact skin. Physiological sources of p-cresol are represented by some gut bacteria which express synthetic enzymes not found in human cells. Urinary p-cresol or its conjugated derivative p-cresylsulfate hold the promise of representing one of these biomarkers in small autistic children and could contribute to identify a subgroup of ASD children characterized by greater severity, to better delineate abnormal gut function in autism and perhaps the entire pathophysiology of the disease in at least some individuals.

2014 - Urinary p-cresol is elevated in young French children with autism spectrum disorder: A replication study [Articolo su rivista]
Gabriele, Stefano; Sacco, Roberto; Cerullo, Sonia; Neri, Cristina; Urbani, Andrea; Tripi, Gabriele; Malvy, Joëlle; Barthelemy, Catherine; Bonnet-Brihault, Frédérique; Persico, Antonio M.
abstract

The aromatic compound p-cresol (4-methylphenol) has been found elevated in the urines of Italian autistic children up to 8 years of age. The present study aims at replicating these initial findings in an ethnically distinct sample and at extending them by measuring also the three components of urinary p-cresol, namely p-cresylsulfate, p-cresylglucuronate and free p-cresol. Total urinary p-cresol, p-cresylsulfate and p-cresylglucuronate were significantly elevated in 33 French autism spectrum disorder (ASD) cases compared with 33 sex- and age-matched controls (p &lt; 0.05). This increase was limited to ASD children aged ≤8 years (p &lt; 0.01), and not older (p = 0.17). Urinary levels of p-cresol and p-cresylsulfate were associated with stereotypic, compulsive/repetitive behaviors (p &lt; 0.05), although not with overall autism severity. These results confirm the elevation of urinary p-cresol in a sizable set of small autistic children and spur interest into biomarker roles for p-cresol and p-cresylsulfate in autism.

2014 - Using genetic findings in autism for the development of new pharmaceutical compounds [Articolo su rivista]
Vorstman, Jacob A. S.; Spooren, Will; Persico, Antonio M.; Collier, David A.; Aigner, Stefan; Jagasia, Ravi; Glennon, Jeffrey C.; Buitelaar, Jan K.
abstract

RATIONALE: The main reason for the current lack of effective treatments for the core symptoms of autism is our limited understanding of the biological mechanisms underlying this heterogeneous group of disorders. A primary value of genetic research is enhancing our insight into the biology of autism through the study of identified autism risk genes. OBJECTIVES: In the current review we discuss (1) the genes and loci that are associated with autism, (2) how these provide us with essential cues as to what neurobiological mechanisms may be involved, and (3) how these mechanisms may be used as targets for novel treatments. Next, we provide an overview of currently ongoing clinical trials registered at clinicaltrials.gov with a variety of compounds. Finally, we review current approaches used to translate knowledge derived from gene discovery into novel pharmaceutical compounds and discuss their pitfalls and problems. CONCLUSIONS: An increasing number of genetic variants associated with autism have been identified. This will generate new ideas about the biological mechanisms involved in autism, which in turn may provide new leads for the development of novel pharmaceutical compounds. To optimize this pipeline of drug discovery, large-scale international collaborations are needed for gene discovery, functional validation of risk genes, and improvement of clinical outcome measures and clinical trial methodology in autism.

2013 - Age-Dependent Decrease and Alternative Splicing of Methionine Synthase mRNA in Human Cerebral Cortex and an Accelerated Decrease in Autism [Articolo su rivista]
Muratore, Christina R.; Hodgson, Nathaniel W.; Trivedi, Malav S.; Abdolmaleky, Hamid M.; Persico, Antonio M.; Lintas, Carla; De La Monte, Suzanne; Deth, Richard C.
abstract

The folate and vitamin B12-dependent enzyme methionine synthase (MS) is highly sensitive to cellular oxidative status, and lower MS activity increases production of the antioxidant glutathione, while simultaneously decreasing more than 200 methylation reactions, broadly affecting metabolic activity. MS mRNA levels in postmortem human cortex from subjects across the lifespan were measured and a dramatic progressive biphasic decrease of more than 400-fold from 28 weeks of gestation to 84 years was observed. Further analysis revealed alternative splicing of MS mRNA, including deletion of folatebinding domain exons and age-dependent deletion of exons from the cap domain, which protects vitamin B12 (cobalamin) from oxidation. Although three species of MS were evident at the protein level, corresponding to full-length and alternatively spliced mRNA transcripts, decreasing mRNA levels across the lifespan were not associated with significant changes in MS protein or methionine levels. MS mRNA levels were significantly lower in autistic subjects, especially at younger ages, and this decrease was replicated in cultured human neuronal cells by treatment with TNF-a, whose CSF levels are elevated in autism. These novel findings suggest that rather than serving as a housekeeping enzyme, MS has a broad and dynamic role in coordinating metabolism in the brain during development and aging. Factors adversely affecting MS activity, such as oxidative stress, can be a source of risk for neurological disorders across the lifespan via their impact on methylation reactions, including epigenetic regulation of gene expression.

2013 - Autism genetics [Articolo su rivista]
Persico, Antonio M; Napolioni, Valerio
abstract

Autism spectrum disorder (ASD) is a severe neuropsychiatric disease with strong genetic underpinnings. However, genetic contributions to autism are extremely heterogeneous, with many different loci underlying the disease to a different extent in different individuals. Moreover, the phenotypic expression (i.e., "penetrance") of these genetic components is also highly variable, ranging from fully penetrant point mutations to polygenic forms with multiple gene-gene and gene-environment interactions. Furthermore, many genes involved in ASD are also involved in intellectual disability, further underscoring their lack of specificity in phenotypic expression. We shall hereby review current knowledge on the genetic basis of ASD, spanning genetic/genomic syndromes associated with autism, monogenic forms due to copy number variants (CNVs) or rare point mutations, mitochondrial forms, and polygenic autisms. Finally, the recent contributions of genome-wide association and whole exome sequencing studies will be highlighted.

2013 - Autisms [Capitolo/Saggio]
Persico, A. M.
abstract

Autism is a complex behavioral disorder with onset during early childhood and typically a life-long course. It is characterized by deficits in communication and social interaction, as well as by stereotypic behaviors and restricted patterns of interest. Its incidence has dramatically risen during the last two decades from 2-5/10,000 to approximately 1-2/1,000 children. The most consistently recognized neurobiological underpinning consists in a relative disconnection between distant brain regions, contrasting with local hyperconnectivity. Autism is increasingly recognized as a heterogeneous mixture of developmental disorders with strong genetic underpinnings. We thus hereby provide a genetically-based classification of the “autisms” distinguishing (a) “classical” syndromic forms, due to known cytogenetic aberrations, copy number variants or mitochondrial DNA abnormalities; (b) recently discovered monogenic autisms, due to highly penetrant mutations affecting synaptic, chromatin architecture, morphogenetic, or calcium-related genes; (c) the most widespread forms of polygenic autisms with “multiple-hit” contributions by common genetic variants as well as by gene-environment interactions; and finally (d) teratogenetic forms, due to environmental exposure to drugs, pollutants or viruses. The recognition of the existence of many different “autisms” is not only rich of heuristic potential for future translational research, but is already starting to influence diagnostic and therapeutic strategies at the present time.

2013 - Headache and comorbidity in children and adolescents [Articolo su rivista]
Bellini, Benedetta; Arruda, Marco; Cescut, Alessandra; Saulle, Cosetta; Persico, Antonello; Carotenuto, Marco; Gatta, Michela; Nacinovich, Renata; Piazza, Fausta Paola; Termine, Cristiano; Tozzi, Elisabetta; Lucchese, Franco; Guidetti, Vincenzo
abstract

Headache is one of the most common neurological symptom reported in childhood and adolescence, leading to high levels of school absences and being associated with several comorbid conditions, particularly in neurological, psychiatric and cardiovascular systems. Neurological and psychiatric disorders, that are associated with migraine, are mainly depression, anxiety disorders, epilepsy and sleep disorders, ADHD and Tourette syndrome. It also has been shown an association with atopic disease and cardiovascular disease, especially ischemic stroke and patent foramen ovale (PFO).

2013 - Plasma cytokine profiling in sibling pairs discordant for autism spectrum disorder [Articolo su rivista]
Napolioni, Valerio; Ober-Reynolds, Benjamin; Szelinger, Szabolcs; Corneveaux, Jason J; Pawlowski, Traci; Ober-Reynolds, Sharman; Kirwan, Janet; Persico, Antonio M; Melmed, Raun D; Craig, David W; Smith, Christopher J; Huentelman, Matthew J
abstract

Converging lines of evidence point to the existence of immune dysfunction in autism spectrum disorder (ASD), which could directly affect several key neurodevelopmental processes. Previous studies have shown higher cytokine levels in patients with autism compared with matched controls or subjects with other developmental disorders. In the current study, we used plasma-cytokine profiling for 25 discordant sibling pairs to evaluate whether these alterations occur within families with ASD.

2013 - Urinary p-cresol in autism spectrum disorder [Articolo su rivista]
Persico, Antonio M.; Napolioni, Valerio
abstract

Autism spectrum disorder (ASD) is a neuropsychiatric disorder with onset during early childhood and life-long consequences in most cases. It is characterized by impairment in social interaction and communication, as well as by restricted patterns of interest and stereotyped behaviors. The etiology of autism is highly heterogeneous, encompassing a large range of genetic and environmental factors. Several lines of evidence suggest that, in addition to broader diagnostic criteria and increased awareness, also a real increase in incidence primarily due to greater gene-environment interactions may also be occurring. Environmental exposure to the organic aromatic compound p-cresol (4-methylphenol) is relatively common and occurs through the skin, as well as the gastrointestinal and respiratory systems. However, the largest and most widespread source of this compound is represented by some gut bacteria which express p-cresol synthesizing enzymes not found in human cells. Urinary p-cresol and its conjugated derivative p-cresylsulfate have been found elevated in an initial sample and recently in a replica sample of autistic children below 8 years of age, where it is associated with female sex, greater clinical severity regardless of sex, and history of behavioral regression. Potential sources of p-cresol excess in ASD, such as gut infection, chronic constipation, antibiotics, abnormal intestinal permeability, and environmental exposure, are being investigated. P-cresol may contribute to worsen autism severity and gut dysfunction, often present in autistic children. It may also contribute to a multibiomarker diagnostic panel useful in small autistic children.

2013 - Urinary polyomavirus infections in neurodevelopmental disorders [Articolo su rivista]
Gentile, Ivan; Altieri, Laura; Lintas, Carla; Sacco, Roberto; Curatolo, Paolo; Benvenuto, Arianna; Muratori, Filippo; Santocchi, Elisa; Bravaccio, Carmela; Lenti, Carlo; Faggioli, Raffaella; Rigardetto, Roberto; Gandione, Marina; Portella, Giuseppe; Zappulo, Emanuela; Borgia, Guglielmo; Persico, Antonio M.
abstract

We have recently reported enhanced frequencies of polyomavirus infection in post-mortem brain tissue of autistic patients compared to controls. To further ex- plore potential contributions to neurodevelopmental disorders by polyomaviruses, we have employed spe- cie-specific TaqMan assays to assess the prevalence and titres of BKV, JCV and SV40 in the urines of 87 patients with autism spectrum disorder, 84 controls matched by sex and age with the autistic sample, 15 subjects with Down syndrome and 13 fragile X indi- viduals. Prevalence rates of urinary BKV infection were significantly greater in Down syndrome and fra- gile X patients compared to autistic and control indi- viduals (P &lt; 0.01). In a large majority of patients who showed the presence of urinary genomes, viral titres resulted significantly higher among Down syndrome patients (P &lt; 0.01) compared to controls, autism spec- trum disorder and fragile X individuals, who did not significantly differ from each other. Our results are consistent with previous evidence supporting ham- pered immunological surveillance and/or immune de- ficits in fragile X and especially in Down syndrome patients.

2012 - Advancing the science of developmental neurotoxicity (DNT): testing for better safety evaluation [Articolo su rivista]
Bal-Price, Anna K; Coecke, Sandra; Costa, Lucio; Crofton, Kevin M; Fritsche, Ellen; Goldberg, Alan; Grandjean, Philippe; Lein, Pamela J; Li, Abby; Lucchini, Roberto; Mundy, William R; Padilla, Stephanie; Persico, Antonio M; Seiler, Andrea E M; Kreysa, Joachim
abstract

2012 - Autism: where genetics meets the immune system [Articolo su rivista]
Persico, Antonio M; Van de Water, Judy; Pardo, Carlos A
abstract

2012 - Cluster analysis of autistic patients based on principal pathogenetic components [Articolo su rivista]
Sacco, Roberto; Lenti, Carlo; Saccani, Monica; Curatolo, Paolo; Manzi, Barbara; Bravaccio, Carmela; Persico, Antonio M.
abstract

We have recently described four principal pathogenetic components in autism: (I) circadian and sensory dysfunction, (II) immune abnormalities, (III) neurodevelopmental delay, and (IV) stereotypic behaviors. Using hierarchical and k-means clustering, the same 245 patients assessed in our principal component analysis can be partitioned into four clusters: (a) 43 (17.6%) have prominent immune abnormalities accompanied by some circadian and sensory issues; (b) 44 (18.0%) display major circadian and sensory dysfunction, with little or no immune symptoms; (c) stereotypies predominate in 75 (31.0%); and (d) 83 (33.9%) show a mixture of all four components, with greater disruptive behaviors and mental retardation. The “immune” component provides the largest contributions to phenotypic variance (P = 2.7 x 10–45), followed by “stereotypic behaviors.” These patient clusters may likely differ in genetic and immune underpinnings, developmental trajectories, and response to treatment.

2012 - Consensus paper: pathological role of the cerebellum in autism [Articolo su rivista]
Fatemi, S Hossein; Aldinger, Kimberly A; Ashwood, Paul; Bauman, Margaret L; Blaha, Charles D; Blatt, Gene J; Chauhan, Abha; Chauhan, Ved; Dager, Stephen R; Dickson, Price E; Estes, Annette M; Goldowitz, Dan; Heck, Detlef H; Kemper, Thomas L; King, Bryan H; Martin, Loren A; Millen, Kathleen J; Mittleman, Guy; Mosconi, Matthew W; Persico, Antonio M; Sweeney, John A; Webb, Sara J; Welsh, John P
abstract

There has been significant advancement in various aspects of scientific knowledge concerning the role of cerebellum in the etiopathogenesis of autism. In the current consensus paper, we will observe the diversity of opinions regarding the involvement of this important site in the pathology of autism. Recent emergent findings in literature related to cerebellar involvement in autism are discussed, including: cerebellar pathology, cerebellar imaging and symptom expression in autism, cerebellar genetics, cerebellar immune function, oxidative stress and mitochondrial dysfunction, GABAergic and glutamatergic systems, cholinergic, dopaminergic, serotonergic, and oxytocin-related changes in autism, motor control and cognitive deficits, cerebellar coordination of movements and cognition, gene-environment interactions, therapeutics in autism, and relevant animal models of autism. Points of consensus include presence of abnormal cerebellar anatomy, abnormal neurotransmitter systems, oxidative stress, cerebellar motor and cognitive deficits, and neuroinflammation in subjects with autism. Undefined areas or areas requiring further investigation include lack of treatment options for core symptoms of autism, vermal hypoplasia, and other vermal abnormalities as a consistent feature of autism, mechanisms underlying cerebellar contributions to cognition, and unknown mechanisms underlying neuroinflammation.

2012 - Genome-wide expression studies in Autism spectrum disorder, Rett syndrome, and Down syndrome [Articolo su rivista]
Lintas, Carla; Sacco, Roberto; Persico, Antonio M.
abstract

Though different in their aetiology, autism spectrum disorder (ASD), Rett syndrome (RTT) and Down syndrome (DS) are three neurodevelopmental disorders sharing significant clinical and neuropathological overlaps. Genome-wide expression studies are reviewed and available datasets from post-mortem brains reanalyzed to identify genes and gene pathways dysregulated in all three disorders. Our results surprisingly converge upon immune, and not neurodevelopmental genes, as the most consistently shared abnormality in genome-wide expression patterns. A dysregulated immune response, accompanied by enhanced oxidative stress and abnormal mitochondrial metabolism seemingly represents the common molecular underpinning of these neurodevelopmental disorders. This conclusion may be important for the definition of pharmacological therapies able to ameliorate clinical symptoms across these disorders.

2012 - Gli autismi [Articolo su rivista]
Persico, Antonio
abstract

Th is article reviews all forms of autisms due to known etiologies, including (a) “classical” syndromic forms, (b) mitochondrial disorders, (c) autisms due to small chromosomal rearrangements (Copy Number Variants), (d) syndromic autisms of monogenic nature due to mutations aff ecting single genes involved in the structure and function of synapses, the packaging of DNA into chromatin, the regulation of cell cycle and morphogenesis, calcium signalling, as well as (e) environmental autisms due to teratogenic drugs, such as valproic acid, misoprostol and thalidomide, or prenatal infections with rubella or cytomegalovirus. Whenever present, signs and symptoms which should spur targetted diagnostic testing are outlined. Progress in our understanding of the biochemical and genetic underpinnings of autism spectrum disorder should pave the path toward pharmacological treatments targeted toward the core symptoms of autism and not merely toward comornid conditions, as suggested by the exciting therapeutic results recently obtained in several animal models of syndromic autisms.

2012 - Gli endofenotipi nel disturbo dello spettro autistico [Articolo su rivista]
Cerullo, S.; Sacco, R.; Persico, A. M.
abstract

Questo lavoro riassume le caratteristiche dei principali endofenotipi nel Disturbo dello Spettro Autistico (DSA). I fratelli/sorelle di individui con autismo hanno un rischio di sviluppare la malattia circa 25 volte maggiore rispetto alla popolazione generale; genitori e fratelli non affetti presentano tratti “sottosoglia” qualitativamente simili a quelli presenti in soggetti con autismo. Questi dati riflettono un alto grado di vulnerabilità genetica che tuttavia ha una base poligenica, eterogenea e complessa nella maggior parte dei pazienti. Descritti per la prima volta nelle malattie psichiatriche da Gottesman e Shields, gli “endofenotipi” possono essere definiti come dei tratti quantitativi familiari ed ereditabili, associati a una malattia complessa. Occupando una posizione intermedia tra genotipo e comportamento sia negli individui affetti sia nei loro familiari non affetti, possono contribuire a colmare la distanza tra fenotipi clinici complessi, come il DSA, ed il livello genetico. Gli endofenotipi nel DSA possono essere raggruppati in sette categorie: biochimici, morfologici, endocrini, immunologici, neurofisiologici, neuropsicologici e comportamentali. Questo approccio potrà risultare utile non solo nell’ambito della ricerca, per spiegare il possibile significato di associazioni genotipo-fenotipo, ma anche nell’ambito clinico per distinguere sottogruppi di pazienti relativamente omogenei per fisiopatologia. Porterà inoltre alla identificazione di biomarcatori in grado di aiutare il clinico a formulare una diagnosi precoce ed una valutazione prognostica individualizzata relativamente alla traiettoria di sviluppo ed alla risposta ai trattamenti riabilitativi, soprattutto nei bambini molto piccoli.

2011 - 2p15-p16.1 microdeletion syndrome: Molecular characterization and association of the OTX1 and XPO1 genes with autism spectrum disorders [Articolo su rivista]
Liu, Xudong; Malenfant, Patrick; Reesor, Chelsea; Lee, Alana; Hudson, Melissa L.; Harvard, Chansonette; Qiao, Ying; Persico, Antonio M.; Cohen, Ira L.; Chudley, Albert E.; Forster-Gibson, Cynthia; Rajcan-Separovic, Evica; Lewis, Me Suzanne; Holden, Jeanette J. A.
abstract

Reports of unrelated individuals with autism spectrum disorder (ASD) and similar clinical features having overlapping de novo interstitial deletions at 2p15–p16.1 suggest that this region harbors a gene(s) important to the development of autism. We molecularly characterized two such deletions, selecting two genes in this region, exportin 1 (XPO1) and orthodenticle homolog 1 (OTX1) for association studies in three North American cohorts (Autism Spectrum Disorder – Canadian American Research Consortium (ASD–CARC), New York, and Autism Genetic Resource Exchange (AGRE)) and one Italian cohort (Societa` Italiana per la Ricerca e la Formazione sull’Autismo (SIRFA)) of families with ASD. In XPO1, rs6735330 was associated with autism in all four cohorts (Po0.05), being significant in ASD–CARC cohorts (P-value following false discovery rate correction for multiple testing (PFDR)¼1.29105), the AGRE cohort (PFDR¼0.0011) and the combined families (PFDR¼2.34109). Similarly, in OTX1, rs2018650 and rs13000344 were associated with autism in ASD–CARC cohorts (PFDR¼8.65107 and 6.07105, respectively), AGRE cohort (PFDR¼0.0034 and 0.015, respectively) and the combined families (PFDR¼2.34109 and 0.00017, respectively); associations were marginal or insignificant in the New York and SIRFA cohorts. A significant association (PFDR¼2.631011) was found for the rs2018650G–rs13000344C haplotype. The above three SNPs were associated with severity of social interaction and verbal communication deficits and repetitive behaviors (P-values o0.01). No additional deletions were identified following screening of 798 ASD individuals. Our results indicate that deletion 2p15–p16.1 is not commonly associated with idiopathic ASD, but represents a novel contiguous gene syndrome associated with a constellation of phenotypic features (autism, intellectual disability, craniofacial/CNS dysmorphology), and that XPO1 and OXT1 may contribute to ASD in 2p15–p16.1 deletion cases and non-deletion cases of ASD mapping to this chromosome region.

2011 - Age- and gender-specific epistasis between ADA and TNF-α influences human life-expectancy [Articolo su rivista]
Napolioni, Valerio; Carpi, Francesco M; Giannì, Paola; Sacco, Roberto; Di Blasio, Luca; Mignini, Fiorenzo; Lucarini, Nazzareno; Persico, Antonio
abstract

Aging is a complex phenotype with multiple determinants but a strong genetic component significantly impacts on survival to extreme ages. The dysregulation of immune responses occurring with increasing age is believed to contribute to human morbidity and mortality. Conversely, some genetic determinants of successful aging might reside in those polymorphisms for the immune system genes regulating immune responses. Here we examined the main effects of single loci and multi-locus interactions to test the hypothesis that the adenosine deaminase (ADA) and tumor necrosis factor alpha (TNF-α) genes may influence human life-expectancy. ADA (22G>A, rs73598374) and TNF-α (-308G>A, rs1800629; -238G>A, rs361525) functional SNPs have been determined for 1071 unrelated healthy individuals from Central Italy (18-106 years old) divided into three gender-specific age classes defined according to demographic information and accounting for the different survivals between sexes: for men (women), the first class consists of individuals<66 years old (<73 years old), the second class of individuals 66-88 years old (73-91 years old), and the third class of individuals>88 years old (>91 years old). Single-locus analysis showed that only ADA 22G>A is significantly associated with human life-expectancy in males (comparison 1 (age class 2 vs. age class 1), O.R. 1.943, P=0.036; comparison 2 (age class 3 vs. age class 2), O.R. 0.320, P=0.0056). Age- and gender-specific patterns of epistasis between ADA and TNF-α were found using Generalized Multifactor Dimensionality Reduction (GMDR). In comparison 1, a significant two-loci interaction occurs in females between ADA 22G>A and TNF-α -238G>A (Sign Test P=0.011). In comparison 2, both two-loci and three-loci interaction are significant associated with increased life-expectancy over 88 years in males. In conclusion, we report that a combination of functional SNPs within ADA and TNF-α genes can influence life-expectancy in a gender-specific manner and that males and females follow different pathways to attain longevity.

2011 - Converging evidence for an association of ATP2B2 allelic variants with autism in male subjects [Articolo su rivista]
Carayol, Jérme; Sacco, Roberto; Tores, Frédéric; Rousseau, Francis; Lewin, Patricia; Hager, Jorg; Persico, Antonio M.
abstract

Background: Autism is a severe developmental disorder, with strong genetic underpinnings. Previous genome-wide scans unveiled a linkage region spanning 3.5 Mb, located on human chromosome 3p25. This region encompasses the ATP2B2 gene, encoding the plasma membrane calcium-transporting ATPase 2 (PMCA2), which extrudes calcium (Ca2) from the cytosol into the extracellular space. Multiple lines of evidence support excessive intracellular Ca2 signaling in autism spectrum disorder (ASD), making ATP2B2 an attractive candidate gene. Methods: We performed a family-based association study in an exploratory sample of 277 autism genetic resource exchange families and in a replication sample including 406 families primarily recruited in Italy. Results: Several markers were significantly associated with ASD in the exploratory sample, and the same risk alleles at single nucleotide polymorphisms rs3774180, rs2278556, and rs241509 were found associated with ASD in the replication sample after correction for multiple testing. In both samples, the association was present in male subjects only. Markers associated with autism are all comprised within a single block of strong linkage disequilibrium spanning several exons, and the “risk” allele seems to follow a recessive mode of transmission. Conclusions: These results provide converging evidence for an association between ATP2B2 gene variants and autism in male subjects, spurring interest into the identification of functional variants, most likely involved in the homeostasis of Ca2 signaling. Additional support comes from a recent genome-wide association study by the Autism Genome Project, which highlights the same linkage disequilibrium region of the gene.

2011 - Family-based association study of ITGB3 in autism spectrum disorder and its endophenotypes [Articolo su rivista]
Napolioni, Valerio; Lombardi, Federica; Sacco, Roberto; Curatolo, Paolo; Manzi, Barbara; Alessandrelli, Riccardo; Militerni, Roberto; Bravaccio, Carmela; Lenti, Carlo; Saccani, Monica; Schneider, Cindy; Melmed, Raun; Pascucci, Tiziana; Puglisi-Allegra, Stefano; Reichelt, Karl-Ludvig; Rousseau, Francis; Lewin, Patricia; Persico, Antonio M
abstract

The integrin-b 3 gene (ITGB3), located on human chromosome 17q21.3, was previously identified as a quantitative trait locus (QTL) for 5-HT blood levels and has been implicated as a candidate gene for autism spectrum disorder (ASD). We performed a family-based association study in 281 simplex and 12 multiplex Caucasian families. ITGB3 haplotypes are significantly associated with autism (HBAT, global P¼0.038). Haplotype H3 is largely over-transmitted to the affected offspring and doubles the risk of an ASD diagnosis (HBAT P¼0.005; odds ratio (OR)¼2.000), at the expense of haplotype H1, which is undertransmitted (HBAT P¼0.018; OR¼0.725). These two common haplotypes differ only at rs12603582 located in intron 11, which reaches a P-value of 0.072 in single-marker FBAT analyses. Interestingly, rs12603582 is strongly associated with pre-term delivery in our ASD patients (P¼0.008). On the other hand, it is SNP rs2317385, located at the 5¢ end of the gene, that significantly affects 5-HT blood levels (Mann–Whitney U-test, P¼0.001; multiple regression analysis, P¼0.010). No gene–gene interaction between ITGB3 and SLC6A4 has been detected. In conclusion, we identify a significant association between a common ITGB3 haplotype and ASD. Distinct markers, located toward the 5¢ and 3¢ ends of the gene, seemingly modulate 5-HT blood levels and autism liability, respectively. Our results also raise interest into ITGB3 influences on feto–maternal immune interactions in autism.

2011 - Genome-wide expression studies in autism-spectrum disorders: moving from neurodevelopment to neuroimmunology [Capitolo/Saggio]
Sacco, R.; Persico, A. M.; Garbett, K. A.; Mirnics, K.
abstract

2011 - Lack of infection with XMRV or other MLV-related viruses in blood, post-mortem brains and paternal gametes of autistic individuals [Articolo su rivista]
Lintas, Carla; Guidi, Francesco; Manzi, Barbara; Mancini, Antonio; Curatolo, Paolo; Persico, Antonio M.
abstract

Background: Autistic spectrum disorder (ASD) is characterized by impaired language, communication and social skills, as well as by repetitive and stereotypic patterns of behavior. Many autistic subjects display a dysregulation of the immune system which is compatible with an unresolved viral infection with prenatal onset, potentially due to vertical viral transmission. Recently, the xenotropic murine leukemia virus-related virus (XMRV) has been implicated in chronic fatigue syndrome (CFS) and in prostate cancer by several, though not all studies. Methodology/Principal Findings: We assessed whether XMRV or other murine leukemia virus (MLV)-related viruses are involved in autistic disorder. Using nested PCR targeted to gag genomic sequences, we screened DNA samples from: (i) peripheral blood of 102 ASD patients and 97 controls, (ii) post-mortem brain samples of 20 ASD patients and 17 sexand age-matched controls, (iii) semen samples of 11 fathers of ASD children, 25 infertile individuals and 7 fertile controls. No XMRV gag DNA sequences were detected, whereas peripheral blood samples of 3/97 (3.1%) controls were positive for MLV. Conclusions|Significance: No MLV-related virus was detected in blood, brain, and semen samples of ASD patients or fathers. Hence infection with XMRV or other MLV-related viruses is unlikely to contribute to autism pathogenesis.

2011 - The mitochondrial aspartate/glutamate carrier AGC1 and calcium homeostasis: Physiological links and abnormalities in autism [Articolo su rivista]
Napolioni, Valerio; Persico, Antonio M.; Porcelli, Vito; Palmieri, Luigi
abstract

Autism spectrum disorder (ASD) is a severe, complex neurodevelopmental disorder characterized by impairments in reciprocal social interaction and communication, and restricted and stereotyped patterns of interests and behaviors. Recent evidence has unveiled an important role for calcium (Ca2+) signaling in the pathogenesis of ASD. Postmortem studies of autistic brains have pointed toward abnormalities in mitochondrial function as possible downstream consequences of altered Ca2+ signaling, abnormal synapse formation, and dysreactive immunity. SLC25A12, an ASD susceptibility gene, encodes the Ca2+-regulated mitochondrial aspartate–glutamate carrier, isoform 1 (AGC1). AGC1 is an important component of the malate/aspartate shuttle, a crucial system supporting oxidative phosphorylation and adenosine triphosphate (ATP) production. Here, we review the physiological roles of AGC1, its links to calcium homeostasis, and its involvement in autism pathogenesis.

2011 - Urinary p-cresol is elevated in small children with severe autism spectrum disorder [Articolo su rivista]
Altieri, Laura; Neri, Cristina; Sacco, Roberto; Curatolo, Paolo; Benvenuto, Arianna; Muratori, Filippo; Santocchi, Elisa; Bravaccio, Carmela; Lenti, Carlo; Saccani, Monica; Rigardetto, Roberto; Gandione, Marina; Urbani, Andrea; Persico, Antonio M.
abstract

Several studies have described in autistic patients an overgrowth of unusual gut bacterial strains, able to push the fermentation of tyrosine up to the formation of p-cresol. We compared levels of urinary p-cresol, measured by highperformance liquid chromatography–ultraviolet, in 59 matched case-control pairs. Urinary p-cresol was significantly elevated in autistic children smaller than 8 years of age (p < 0.01), typically females (p < 0.05), and more severely affected regardless of sex (p < 0.05). Urinary cotinine measurements excluded smoking-related hydrocarbon contaminations as contributors to these differences. Hence, elevated urinary p-cresol may serve as a biomarker of autism liability in small children, especially females and more severely affected males.

2010 - Altered calcium homeostasis in autism-spectrum disorders: Evidence from biochemical and genetic studies of the mitochondrial aspartate/glutamate carrier AGC1 [Articolo su rivista]
Palmieri, L.; Papaleo, V.; Porcelli, V.; Scarcia, P.; Gaita, L.; Sacco, R.; Hager, J.; Rousseau, F.; Curatolo, P.; Manzi, B.; Militerni, R.; Bravaccio, C.; Trillo, S.; Schneider, C.; Melmed, R.; Elia, M.; Lenti, C.; Saccani, M.; Pascucci, T.; Puglisi-Allegra, S.; Reichelt, K. -L.; Persico, A. M.
abstract

Autism is a severe developmental disorder, whose pathogenetic underpinnings are still largely unknown. Temporocortical gray matter from six matched patient–control pairs was used to perform post-mortem biochemical and genetic studies of the mitochondrial aspartate/ glutamate carrier (AGC), which participates in the aspartate/malate reduced nicotinamide adenine dinucleotide shuttle and is physiologically activated by calcium (Ca2þ). AGC transport rates were significantly higher in tissue homogenates from all six patients, including those with no history of seizures and with normal electroencephalograms prior to death. This increase was consistently blunted by the Ca2þ chelator ethylene glycol tetraacetic acid; neocortical Ca2þ levels were significantly higher in all six patients; no difference in AGC transport rates was found in isolated mitochondria from patients and controls following removal of the Ca2þ-containing postmitochondrial supernatant. Expression of AGC1, the predominant AGC isoform in brain, and cytochrome c oxidase activity were both increased in autistic patients, indicating an activation of mitochondrial metabolism. Furthermore, oxidized mitochondrial proteins were markedly increased in four of the six patients. Variants of the AGC1-encoding SLC25A12 gene were neither correlated with AGC activation nor associated with autism-spectrum disorders in 309 simplex and 17 multiplex families, whereas some unaffected siblings may carry a protective gene variant. Therefore, excessive Ca2þ levels are responsible for boosting AGC activity, mitochondrial metabolism and, to a more variable degree, oxidative stress in autistic brains. AGC and altered Ca2þ homeostasis play a key interactive role in the cascade of signaling events leading to autism: their modulation could provide new preventive and therapeutic strategies.

2010 - Association of autism with polyomavirus infection in postmortem brains [Articolo su rivista]
Lintas, Carla; Altieri, Laura; Lombardi, Federica; Sacco, Roberto; Persico, Antonio M.
abstract

Autism is a highly heritable behavioral disorder. Yet, two decades of genetic investigation have unveiled extremely few cases that can be solely explained on the basis of de novo mutations or cytogenetic abnormalities. Vertical viral transmission represents a nongenetic mechanism of disease compatible with high parent-to-offspring transmission and with low rates of disease-specific genetic abnormalities. Vertically transmitted viruses should be found more frequently in the affected tissues of autistic individuals compared to controls. Our initial step was thus to assess by nested polymerase chain reaction (PCR) and DNA sequence analysis the presence of cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus type 1 (HSV1), herpes simplex virus type 2 (HSV2), human herpes virus 6 (HHV6), BK virus (BKV), JC virus (JCV), and simian virus 40 (SV40) in genomic DNA extracted from postmortem temporocortical tissue (Brodmann areas 41/42) belonging to 15 autistic patients and 13 controls. BKV, JCV, and SV40 combined are significantly more frequent among autistic patients compared to controls (67% versus 23%, respectively; P < .05). The majority of positives yielded archetypal sequences, whereas six patients and two controls unveiled single–base pair changes in two or more sequenced clones. No association is present with the remaining viruses, which are found in relatively few individuals (N £ 3). Also polyviral infections tend to occur more frequently in the brains of autistic patients compared to controls (40% versus 7.7%, respectively; P = .08). Follow-up studies exploring vertical viral transmission as a possible pathogenetic mechanism in autistic disorder should focus on, but not be limited to, the role of polyomaviruses.

2010 - Candidate gene study of HOXB1 in autism spectrum disorder [Articolo su rivista]
Muscarella, Lucia A.; Guarnieri, Vito; Sacco, Roberto; Curatolo, Paolo; Manzi, Barbara; Alessandrelli, Riccardo; Giana, Grazia; Militerni, Roberto; Bravaccio, Carmela; Lenti, Carlo; Saccani, Monica; Schneider, Cindy; Melmed, Raun; D'Agruma, Leonardo; Persico, Antonio M.
abstract

Background: HOXB1 plays a major role in brainstem morphogenesis and could partly determine the cranial circumference in conjunction with HOXA1. In our sample, HOXA1 alleles significantly influence head growth rates both in autistic patients and in population controls. An initial report, suggesting that HOXB1 could confer autism vulnerability in interaction with HOXA1, was not confirmed by five small association studies. Methods: Our sample includes 269 autistic individuals, belonging to 219 simplex and 28 multiplex families. A mutational analysis of the two exons and flanking intronic sequences of the HOXB1 gene was carried out in 84 autistic patients by denaturing high performance liquid chromatography, followed by DNA sequencing. Identified rare variants were then searched by a restriction analysis in 236 autistic patients and 325-345 controls. Case-control and family-based association studies were performed on two common variants in 169 Italian patients versus 184 Italian controls and in 247 trios. Results: We identified three common polymorphisms, rs72338773 [c.82insACAGCGCCC (INS/nINS)], rs12939811 [c.309A&gt;T (Q103H)], and rs7207109 [c.450G&gt;A (A150A)] and three rare variants, namely IVS1+63G&gt;A, rs35115415 [c.702G&gt;A (V234V)] and c.872_873delinsAA (S291N). SNPs rs72338773 and rs12939811 were not associated with autism, using either a case-control (alleles, exact P = 0.13) or a family-based design transmission/disequilibrium test (TDT)χ2 = 1.774, P = 0.183]. The rare variants, all inherited from one of the parents, were present in two Italian and in two Caucasian-American families. Autistic probands in two families surprisingly inherited a distinct rare variant from each parent. The IVS1+63A allele was present in 3/690 control chromosomes, whereas rare alleles at rs35115415 and c.872_873delinsAA (S291N) were not found in 662 and 650 control chromosomes, respectively. The INS-T309 allele influenced head size, but its effect appears more modest and shows no interaction with HOXA1 alleles. The INS-T309 allele is also associated with more severe stereotypic behaviours, according to ADI-R scores (N = 60 patients, P &lt; 0.01). Conclusions: HOXB1 mutations do not represent a common cause of autism, nor do HOXB1 common variants play important roles in autism vulnerability. HOXB1 provides minor, albeit detectable contributions to head circumference in autistic patients, with HOXA1 displaying more prominent effects. HOXB1 variants may modulate the clinical phenotype, especially in the area of stereotypic behaviours.

2010 - Decreased serum arylesterase activity in autism spectrum disorders [Articolo su rivista]
Gaita, Laura; Manzi, Barbara; Sacco, Roberto; Lintas, Carla; Altieri, Laura; Lombardi, Federica; Pawlowski, Tracy L.; Redman, Margot; Craig, David W.; Huentelman, Matthew J.; Ober-Reynolds, Sharman; Brautigam, Sarah; Melmed, Raun; Smith, Christopher J.; Marsillach, Judith; Camps, Jordi; Curatolo, Paolo; Persico, Antonio M.
abstract

The PON1 gene, previously found associated with autism spectrum disorders (ASDs), encodes a serum protein responsible for the detoxification of organophosphates (OPs) and able to exert several enzymatic activities. PON1 arylesterase, but not diazoxonase activity, was significantly decreased in 174 ASD patients compared to 175 first-degree relatives and 144 controls (P=2.65×10−16). First degree relatives displayed intermediate activities, closer to patient than to control levels. Differences between patients, first-degree relatives and controls were especially evident among 164 Italians compared to 329 Caucasian-Americans, because arylesterase activity was significantly higher in Italian controls, compared to Caucasian-American controls (P=2.84×10−16). Arylesterase activity and PON protein concentrations were not significantly correlated, supporting a functional inhibition of arylesterase activity in ASD patients over quantitative changes in protein amounts. Serum arylesterase activity, in combination with PON1 genotypes at two single nucleotide polymorphisms (SNPs) known to influence protein amounts (rs705379: C-108T) and substrate specificity (rs662: Q192R), was able to discriminate ASD patients from controls with elevated sensitivity and specificity, depending on genotype and ethnic group. Serum arylesterase activity and genotyping at these two SNPs could thus represent an informative biochemical/genetic test, able to aid clinicians in estimating autism risk in ethnic groups with higher baseline arylesterase activity levels.

2010 - Developmental roles of the serotonin transporter [Capitolo/Saggio]
Persico, A. M.
abstract

From invertebrates to humans, serotonin (5-HT) exerts structural effects especially during development. The 5-HT transporter (SERT) directly regulates these effects by maintaining extracellular 5-HT concentrations within a physiological range and possibly by modulating the intracellular redox state of the cell. This chapter addresses 5-HT trophic effects on developing neural and non-neural mammalian cells, and summarizes SERT roles in 5HT-mediated structural effects from basic neurodevelopment to human teratology.

2010 - Mitochondrial dysfunction in autism spectrum disorders: Cause or effect? [Articolo su rivista]
Palmieri, Luigi; Persico, Antonio M.
abstract

Autism Spectrum Disorders encompass severe developmental disorders characterized by variable degrees of impairment in language, communication and social skills, as well as by repetitive and stereotypic patterns of behaviour. Substantial percentages of autistic patients display peripheral markers of mitochondrial energy metabolism dysfunction, such as (a) elevated lactate, pyruvate, and alanine levels in blood, urine and/or cerebrospinal fluid, (b) serum carnitine deficiency, and/or (c) enhanced oxidative stress. These biochemical abnormalities are accompanied by highly heterogeneous clinical presentations, which generally (but by no means always) encompass neurological and systemic symptoms relatively unusual in idiopathic autistic disorder. In some patients, these abnormalities have been successfully explained by the presence of specific mutations or rearrangements in their mitochondrial or nuclear DNA. However, in the majority of cases, abnormal energy metabolism cannot be immediately linked to specific genetic or genomic defects. Recent evidence from post-mortem studies of autistic brains points toward abnormalities in mitochondrial function as possible downstream consequences of dysreactive immunity and altered calcium (Ca2+) signalling.

2010 - Neocortical RELN promoter methylation increases significantly after puberty [Articolo su rivista]
Lintas, Carla; Persico, Antonio M
abstract

Reelin plays a pivotal role in neurodevelopment. Excessive RELN promoter methylation and/or decreased RELN gene expression have been described in schizophrenia and autism. We assessed RELN promoter methylation in post-mortem temporocortical tissue (Brodmann Area 41/42) of three prepuberal and six postpuberal normal individuals. The former display very little or no methylation, whereas most postpuberal individuals are heavily methylated, especially at CpG positions located between -131 and -98 bp (prepuberal vs. postpuberal, P<0.05). Sex hormones thus seemingly boost DNA methylation at the RELN promoter. This physiological change could significantly contribute to the onset of schizophrenia and the worsening of autistic behaviors, both typically occurring at puberty.

2010 - Principal pathogenetic components and biological endophenotypes in autism spectrum disorders [Articolo su rivista]
Sacco, Roberto; Curatolo, Paolo; Manzi, Barbara; Militerni, Roberto; Bravaccio, Carmela; Frolli, Alessandro; Lenti, Carlo; Saccani, Monica; Elia, Maurizio; Reichelt, Karl-Ludvig; Pascucci, Tiziana; Puglisi-Allegra, Stefano; Persico, Antonio M.
abstract

Autism is a complex neurodevelopmental disorder, likely encompassing multiple pathogenetic components. The aim of this study is to begin identifying at least some of these components and to assess their association with biological endophenotypes. To address this issue, we recruited 245 Italian patients with idiopathic autism spectrum disorders and their first-degree relatives. Using a stepwise approach, patient and family history variables were analyzed using principal component analysis (‘‘exploratory phase’’), followed by intra- and inter-component cross-correlation analyses (‘‘followup phase’’), and by testing for association between each component and biological endophenotypes, namely head circumference, serotonin blood levels, and global urinary peptide excretion rates (‘‘biological correlation phase’’). Four independent components were identified, namely ‘‘circadian & sensory dysfunction,’’ ‘‘immune dysfunction,’’ ‘‘neurodevelopmental delay,’’ and ‘‘stereotypic behavior,’’ together representing 74.5% of phenotypic variance in our sample. Marker variables in the latter three components are positively associated with macrocephaly, global peptiduria, and serotonin blood levels, respectively. These four components point toward at least four processes associated with autism, namely (I) a disruption of the circadian cycle associated with behavioral and sensory abnormalities, (II) dysreactive immune processes, surprisingly linked both to prenatal obstetric complications and to excessive postnatal body growth rates, (III) a generalized developmental delay, and (IV) an abnormal neural circuitry underlying stereotypies and early social behaviors.

2010 - Reply to the letter to the editor: Polyomaviruses and autism: More than simple association? [Recensione in Rivista]
Persico, Antonio M.
abstract

2009 - Autism-associated functional polymorphisms in SLC25A12 gene [Brevetto]
Persico, A. M.; Lombardi, F.
abstract

The present invention relates to a method of determining the presence of or predisposition to autism, or to an autism spectrum disorder in a subject, the method comprising detecting the presence of single nucleotide polymorphism (SNP) rs3765166 and/or rsll757 of SLC25A12 gene, in a sample from said subjects.

2009 - Autism-associated functional polymorphisms in SLC25A12 gene [Brevetto]
Persico, A. M.; Lombardi, F.
abstract

The present invention relates to a method of determining the presence of or predisposition to autism, or to an autism spectrum disorder in a subject, the method comprising detecting the presence of single nucleotide polymorphism (SNP) rs3765166 and/or rsll757 of SLC25A12 gene, in a sample from said subjects.

2009 - Autistic phenotypes and genetic testing: State-of-the-art for the clinical geneticist [Articolo su rivista]
Lintas, C.; Persico, A. M.
abstract

Autism spectrum disorders represent a group of developmental disorders with strong genetic underpinnings. Several cytogenetic abnormalities or de novo mutations able to cause autism have recently been uncovered. In this study, the literature was reviewed to highlight genotype–phenotype correlations between causal gene mutations or cytogenetic abnormalities and behavioural or morphological phenotypes. Based on this information, a set of practical guidelines is proposed to help clinical geneticists pursue targeted genetic testing for patients with autism whose clinical phenotype is suggestive of a specific genetic or genomic aetiology.

2009 - Genomic and epigenetic evidence for oxytocin receptor deficiency in autism [Articolo su rivista]
Gregory, S. G.; Connelly, J. J.; Towers, A. J.; Johnson, J.; Biscocho, D.; Markunas, C. A.; Lintas, C.; Abramson, R. K.; Wright, H. H.; Ellis, P.; Langford, C. F.; Worley, G.; Delong, G. R.; Murphy, S. K.; Cuccaro, M. L.; Persico, A.; Pericak-Vance, M. A.
abstract

Background: Autism comprises a spectrum of behavioral and cognitive disturbances of childhood development and is known to be highly heritable. Although numerous approaches have been used to identify genes implicated in the development of autism, less than 10% of autism cases have been attributed to single gene disorders. Methods: We describe the use of high-resolution genome-wide tilepath microarrays and comparative genomic hybridization to identify copy number variants within 119 probands from multiplex autism families. We next carried out DNA methylation analysis by bisulfite sequencing in a proband and his family, expanding this analysis to methylation analysis of peripheral blood and temporal cortex DNA of autism cases and matched controls from independent datasets. We also assessed oxytocin receptor (OXTR) gene expression within the temporal cortex tissue by quantitative real-time polymerase chain reaction (PCR). Results: Our analysis revealed a genomic deletion containing the oxytocin receptor gene, OXTR (MIM accession no.: 167055), previously implicated in autism, was present in an autism proband and his mother who exhibits symptoms of obsessive-compulsive disorder. The proband's affected sibling did not harbor this deletion but instead may exhibit epigenetic misregulation of this gene through aberrant gene silencing by DNA methylation. Further DNA methylation analysis of the CpG island known to regulate OXTR expression identified several CpG dinucleotides that show independent statistically significant increases in the DNA methylation status in the peripheral blood cells and temporal cortex in independent datasets of individuals with autism as compared to control samples. Associated with the increase in methylation of these CpG dinucleotides is our finding that OXTR mRNA showed decreased expression in the temporal cortex tissue of autism cases matched for age and sex compared to controls. Conclusion: Together, these data provide further evidence for the role of OXTR and the oxytocin signaling pathway in the etiology of autism and, for the first time, implicate the epigenetic regulation of OXTR in the development of the disorder. © 2009 Gregory et al; licensee BioMed Central Ltd.

2009 - Involvement of the PRKCB1 gene in autistic disorder: Significant genetic association and reduced neocortical gene expression [Articolo su rivista]
Lintas, C.; Sacco, R.; Garbett, K.; Mirnics, K.; Militerni, R.; Bravaccio, C.; Curatolo, P.; Manzi, B.; Schneider, C.; Melmed, R.; Elia, M.; Pascucci, T.; Puglisi-Allegra, S.; Reichelt, K. -L.; Persico, A. M.
abstract

Protein kinase C enzymes play an important role in signal transduction, regulation of gene expression and control of cell division and differentiation. The fsI and bII isoenzymes result from the alternative splicing of the PKCb gene (PRKCB1), previously found to be associated with autism. We performed a family-based association study in 229 simplex and 5 multiplex families, and a postmortem study of PRKCB1 gene expression in temporocortical gray matter (BA41/42) of 11 autistic patients and controls. PRKCB1 gene haplotypes are significantly associated with autism (P < 0.05) and have the autistic endophenotype of enhanced oligopeptiduria (P < 0.05). Temporocortical PRKCB1 gene expression was reduced on average by 35 and 31% for the PRKCB1-1 and PRKCB1-2 isoforms (P < 0.01 and < 0.05, respectively) according to qPCR. Protein amounts measured for the PKCbII isoform were similarly decreased by 35% (P= 0.05). Decreased gene expression characterized patients carrying the ‘normal’ PRKCB1 alleles, whereas patients homozygous for the autism-associated alleles displayed mRNA levels comparable to those of controls. Whole genome expression analysis unveiled a partial disruption in the coordinated expression of PKCb-driven genes, including several cytokines. These results confirm the association between autism and PRKCB1 gene variants, point toward PKCb roles in altered epithelial permeability, demonstrate a significant downregulation of brain PRKCB1 gene expression in autism and suggest that it could represent a compensatory adjustment aimed at limiting an ongoing dysreactive immune process. Altogether, these data underscore potential PKCb roles in autism pathogenesis and spur interest in the identification and functional characterization of PRKCB1 gene variants conferring autism vulnerability.

2008 - Genetic evidence implicating multiple genes in the met receptor tyrosine kinase pathway in autism spectrum disorder [Articolo su rivista]
Campbell, Daniel B.; Li, Chun; Sutcliffe, James S.; Persico, Antonio M.; Levitt, Pat
abstract

A functional promoter variant of the gene encoding the MET receptor tyrosine kinase alters SP1 and SUB1 transcription factor binding, and is associated with autism spectrum disorder (ASD). Recent analyses of postmortem cerebral cortex from ASD patients revealed altered expression of MET protein and three transcripts encoding proteins that regulate MET signaling, hepatocyte growth factor (HGF), urokinase plasminogen activator receptor (PLAUR) and plasminogen activator inhibitor-1 (SERPINE1). To address potential risk conferred by multiple genes in the MET signaling pathway, we screened all exons and 5′ promoter regions for variants in the five genes encoding proteins that regulate MET expression and activity. Identified variants were genotyped in 664 families (2,712 individuals including 1,228 with ASD) and 312 unrelated controls. Replicating our initial findings, family-based association test (FBAT) analyses demonstrated that the MET promoter variant rs1858830 C allele was associated with ASD in 101 new families (P=0.033). Two other genes in the MET signaling pathway also may confer risk. A haplotype of the SERPINE1 gene exhibited significant association. In addition, the PLAUR promoter variant rs344781 T allele was associated with ASD by both FBAT (P=0.006) and case-control analyses (P=0.007). The PLAUR promoter rs344781 relative risk was 1.93 (95% Confidence Interval [CI]: 1.12−3.31) for genotype TT and 2.42 (95% CI: 1.38−4.25) for genotype CT compared to genotype CC. Gene-gene interaction analyses suggested a significant interaction between MET and PLAUR. These data further support our hypothesis that genetic susceptibility impacting multiple components of the MET signaling pathway contributes to ASD risk.

2008 - Immune transcriptome alterations in the temporal cortex of subjects with autism [Articolo su rivista]
Garbett, Krassimira; Ebert, Philip J.; Mitchell, Amanda; Lintas, Carla; Manzi, Barbara; Mirnics, Károly; Persico, Antonio M.
abstract

Autism is a severe disorder that involves both genetic and environmental factors. Expression profiling of the superior temporal gyrus of six autistic subjects and matched controls revealed increased transcript levels of many immune system-related genes. We also noticed changes in transcripts related to cell communication, differentiation, cell cycle regulation and chaperone systems. Critical expression changes were confirmed by qPCR (BCL6, CHI3L1, CYR61, IFI16, IFITM3, MAP2K3, PTDSR, RFX4, SPP1, RELN, NOTCH2, RIT1, SFN, GADD45B, HSPA6, HSPB8 and SERPINH1). Overall, these expression patterns appear to be more associated with the late recovery phase of autoimmune brain disorders, than with the innate immune response characteristic of neurodegenerative diseases.Moreover, a variance-based analysis revealed much greater transcript variability in brains from autistic subjects compared to the control group, suggesting that these genes may represent autism susceptibility genes and should be assessed in follow-up genetic studies.

2008 - Measurement of arylesterase enzymatic activity and assessment of genetic polymorphisms located in the PON1 gene as a diagnostic tool in autism-spectrum disorders [Brevetto]
Persico, A. M.; Sacco, R.; Lintas, C.
abstract

2008 - Reelin gene polymorphisms in autistic disorder [Capitolo/Saggio]
Lintas, C.; Persico, A. M.
abstract

2007 - Altered neocortical cell density and layer thickness in serotonin transporter knockout mice: A quantitation study [Articolo su rivista]
Altamura, C.; Dell'Acqua, M. L.; Moessner, R.; Murphy, D. L.; Lesch, K. P.; Persico, Antonio M.
abstract

The neurotransmitter serotonin (5-HT) plays morphogenetic roles during development, and their alteration could contribute to autism pathogenesis in humans. To further characterize 5-HT contributions to neocortical development, we assessed the thickness and neuronal cell density of various cerebral cortical areas in 5-HT transporter (5-HTT) knockout (ko) mice, characterized by elevated extracellular 5-HT levels. The thickness of layer IV is decreased in 5-HTT knockout mice compared to wildtype mice. The overall effect on cortical thickness, however, depends on the genetic background of the mice. Overall cortical thickness is decreased in many cortical areas of 5-HTT ko mice with a mixed c129-CD1-C57BL/6J background. Instead, 5-HTT ko mice backcrossed into the C57BL/6J background display increases in supragranular and infragranular layers, which compensate entirely for decreased layer IV thickness, resulting in unchanged, or even enhanced cortical thickness. Moreover, significant increases in neuronal cell density are found in 5-HTT ko mice with a C57BL/6J background (wt:hz:ko ratio = 1.00:1.04:1.17), but not in mixed c129-CD1-C57BL/6J 5-HTT ko animals. These results provide evidence of 5-HTT gene effects on neocortical morphology in epistatic interaction with genetic variants at other loci, and may model the effect of functional 5-HTT gene variants on neocortical development in autism.

2007 - Case-control and family-based association studies of candidate genes in autistic disorder and its endophenotypes: TPH2 and GLO1 [Articolo su rivista]
Sacco, Roberto; Papaleo, Veruska; Hager, Jorg; Rousseau, Francis; Moessner, Rainald; Militerni, Roberto; Bravaccio, Carmela; Trillo, Simona; Schneider, Cindy; Melmed, Raun; Elia, Maurizio; Curatolo, Paolo; Manzi, Barbara; Pascucci, Tiziana; Puglisi-Allegra, Stefano; Reichelt, Karl-Ludvig; Persico, Antonio M.
abstract

Background: The TPH2 gene encodes the enzyme responsible for serotonin (5-HT) synthesis in the Central Nervous System (CNS). Stereotypic and repetitive behaviors are influenced by 5-HT, and initial studies report an association of TPH2 alleles with childhood-onset obsessive-compulsive disorder (OCD) and with autism. GLO1 encodes glyoxalase I, the enzyme which detoxifies α-oxoaldehydes such as methylglyoxal in all living cells. The A111E GLO1 protein variant, encoded by SNP C419A, was identifed in autopsied autistic brains and proposed to act as an autism susceptibility factor. Hyperserotoninemia, macrocephaly, and peptiduria represent some of the best-characterized endophenotypes in autism research. Methods: Family-based and case-control association studies were performed on clinical samples drawn from 312 simplex and 29 multiplex families including 371 non-syndromic autistic patients and 156 unaffected siblings, as well as on 171 controls. TPH2 SNPs rs4570625 and rs4565946 were genotyped using the TaqMan assay; GLO1 SNP C419A was genotyped by PCR and allele-specific restriction digest. Family-based association analyses were performed by TDT and FBAT, case-control by χ2, endophenotypic analyses for 5-HT blood levels, cranial circumference and urinary peptide excretion rates by ANOVA and FBAT. Results: TPH2 alleles and haplotypes are not significantly associated in our sample with autism (rs4570625: TDT P =0.27, and FBAT P = 0.35; rs4565946: TDT P = 0.45, and FBAT P = 0.55; haplotype P = 0.84), with any endophenotype, or with the presence/absence of prominent repetitive and stereotyped behaviors (motor stereotypies: P = 0.81 and 0.84, verbal stereotypies: P = 0.38 and 0.73 for rs4570625 and rs4565946, respectively). Also GLO1 alleles display no association with autism (191 patients vs 171 controls, P = 0.36; TDT P = 0.79, and FBAT P = 0.37), but unaffected siblings seemingly carry a protective gene variant marked by the A419 allele (TDT P &lt; 0.05; patients vs unaffected siblings TDT and FBAT P &lt; 0.00001). Conclusion: TPH2 gene variants are unlikely to contribute to autism or to the presence/absence of prominent repetitive behaviors in our sample, although an influence on the intensity of these behaviors in autism cannot be excluded. GLO1 gene variants do not confer autism vulnerability in this sample, but allele A419 apparently carries a protective effect, spurring interest into functional correlates of the C419A SNP.

2007 - Clinical, Morphological, and Biochemical Correlates of Head Circumference in Autism [Articolo su rivista]
Sacco, Roberto; Militerni, Roberto; Frolli, Alessandro; Bravaccio, Carmela; Gritti, Antonella; Elia, Maurizio; Curatolo, Paolo; Manzi, Barbara; Trillo, Simona; Lenti, Carlo; Saccani, Monica; Schneider, Cindy; Melmed, Raun; Reichelt, Karl-Ludvig; Pascucci, Tiziana; Puglisi-Allegra, Stefano; Persico, Antonio M.
abstract

Background: Head growth rates are often accelerated in autism. This study is aimed at defining the clinical, morphological, and biochemical correlates of head circumference in autistic patients. Methods: Fronto-occipital head circumference was measured in 241 nonsyndromic autistic patients, 3 to 16 years old, diagnosed according to DSM-IV criteria. We assessed 1) clinical parameters using the Autism Diagnostic Observation Schedule, Autism Diagnostic Interview-Revised, Vineland Adaptive Behavioral Scales, intelligence quotient measures, and an ad hoc clinical history questionnaire; 2) height and weight; 3) serotonin (5-HT) blood levels and peptiduria. Results: The distribution of cranial circumference is significantly skewed toward larger head sizes (p .00001). Macrocephaly (i.e., head circumference97th percentile) is generally part of a broader macrosomic endophenotype, characterized by highly significant correlations between head circumference, weight, and height (p .001). A head circumference 75th percentile is associated with more impaired adaptive behaviors and with less impairment in IQ measures and motor and verbal language development. Surprisingly, larger head sizes are significantly associated with a positive history of allergic/immune disorders both in the patient and in his/her first-degree relatives. Conclusions: Our study demonstrates the existence of a macrosomic endophenotype in autism and points toward pathogenetic links with immune dysfunctions that we speculate either lead to or are associated with increased cell cycle progression and/or decreased apoptosis.

2007 - Disruption of cerebral cortex MET signaling in autism spectrum disorder [Articolo su rivista]
Campbell, Daniel B.; D'Oronzio, Rosanna; Garbett, Krassi; Ebert, Philip J.; Mirnics, Karoly; Levitt, Pat; Persico, Antonio M.
abstract

Objective: Multiple genes contribute to autism spectrum disorder (ASD) susceptibility. One particularly promising candidate is the MET gene, which encodes a receptor tyrosine kinase that mediates hepatocyte growth factor (HGF) signaling in brain circuit formation, immune function, and gastrointestinal repair. The MET promoter variant rs1858830 allele “C” is strongly associated with ASD and results in reduced gene transcription. Here we examined expression levels of MET and members of the MET signaling pathway in postmortem cerebral cortex from ASD cases and healthy control subjects. Methods: Protein, total RNA, and DNA were extracted from postmortem temporal cortex gray matter samples (BA 41/42, 52, or 22) belonging to eight pairs of ASD cases and matched control subjects. MET protein expression was determined by Western blotting; messenger RNA expression of MET and other related transcripts was assayed by microarray and quantitative reverse transcriptase polymerase chain reaction. Results: MET protein levels were significantly decreased in ASD cases compared with control subjects. This was accompanied in ASD brains by increased messenger RNA expression for proteins involved in regulating MET signaling activity. Analyses of coexpression of MET and HGF demonstrated a positive correlation in control subjects that was disrupted in ASD cases. Interpretation: Altered expression of MET and related molecules suggests dysregulation of signaling that may contribute to altered circuit formation and function in ASD. The complement of genes that encode proteins involved in MET activation appears to undergo long-term compensatory changes in expression that may be a hallmark contribution to the pathophysiology of ASD.

2007 - HOXA1 gene variants influence head growth rates in humans [Articolo su rivista]
Muscarella, Lucia Anna; Guarnieri, Vito; Sacco, Roberto; Militerni, Roberto; Bravaccio, Carmela; Trillo, Simona; Schneider, Cindy; Melmed, Raun; Elia, Maurizio; Mascia, Maria Lucia; Rucci, Emanuela; Piemontese, Maria Rosaria; D'Agruma, Leonardo; Persico, Antonio M.
abstract

We previously described a significant association between the HOXA1 G218 allele and increased head circumference in autism [Conciatori et al. (2004); Biol Psychiatry 55:413–419]. The present study reveals identical effects also in normal children. HOXA1 A218G alleles and sex explain as much as 10.9 and 6.8% of the variance in head circumference in 142 pediatric controls and in 191 autistic children, aged 3–16 years (F¼6.777, 3 and 141 df,P<0.001 and F¼5.588, 3 and 190 df,P<0.01, respectively). Instead, no association is found in 183 adult controls and in 35 pediatric fragile-X patients. Therefore HOXA1 A218G alleles significantly influence head growth rates, but not final head size, in normal human development. This influence does not differ between normal and autistic children, whereas the lack of FMRP seemingly overwhelms HOXA1 effects in fragile-X patients.

2007 - Measurement of arylesterase enzymatic activity and assessment of genetic polymorphisms located in the PON1 gene as a diagnostic tool in autism-spectrum disorders [Brevetto]
Persico, A. M.; Sacco, R.; Lintas, C.
abstract

The present invention concerns a method for detecting the presence of or predisposition to autism, an autism spectrum disorder, or an autism-associated disorder in a subject, the method comprising measuring an arylesterase enzymatic activity in a sample from the subject, optionally combined with the determination of alleles of PON1 polymorphisms.

2007 - Transmission disequilibrium study of an oligodendrocyte and myelin glycoprotein gene allele in 431 families with an autistic proband [Articolo su rivista]
Martin, Isabelle; Gauthier, Julie; D'Amelio, Marcello; Védrine, Sylviane; Vourc'H, Patrick; Rouleau, Guy A.; Persico, Antonio M.; Andres, Christian R.
abstract

Autistic disorder is a neurodevelopmental disorder where genetic factors play an important role. We previously described an association between a subgroup of French autistic patients and an allele of a non-synonymous single nucleotide polymorphism (nsSNP: OMGP62 G > A or rs11080149) in the gene coding for the oligodendrocyte and myelin glycoprotein (OMG), located at 7 Mb from the marker D17S250, linked to autism in two independent genome scan studies. We report a study on 431 families with 1 affected child from different origins: French Canada (n = 262), Italy (n = 123) and United States (n = 46). We analyzed the transmission of the rs11080149 alleles from parents to their affected children. There was a preferential transmission of the G allele from parents to affected children ( p = 0.0017) in the overall sample. Paternal and maternal transmission rates were both skewed. Taking into account our previous results obtained in a French group of patients, where we observed an association with allele A, a direct role of this polymorphism is improbable in autism. The associations observed in Japanese and French patients, the linkage studies and the present work speak in favor of the existence of a susceptibility gene for autism in the NF1 locus.

2006 - A genetic variant that disrupts MET transcription is associated with autism [Articolo su rivista]
Campbell, Daniel B.; Sutcliffe, James S.; Ebert, Philip J.; Militerni, Roberto; Bravaccio, Carmela; Trillo, Simona; Elia, Maurizio; Schneider, Cindy; Melmed, Raun; Sacco, Roberto; Persico, Antonio M.; Levitt, Pat
abstract

There is strong evidence for a genetic predisposition to autism, and an intense interest in discovering heritable risk factors that disrupt gene function. Based on neurobiological findings and location within a chromosome 7q31 autism candidate gene region, we analyzed the gene encoding the pleiotropic MET receptor tyrosine kinase in a family-based study of autism including 1,231 cases. MET signaling participates in neocortical and cerebellar growth and maturation, immune function and gastrointestinal repair, consistent with reported medical complications in some children with autism. Here we show genetic association (P=0.0005) of a common ‘C’ allele in the promoter region of the MET gene in 204 autism families. The allelic association at this MET variant was confirmed in a replication sample of 539 autism families (P=0.001) and in the combined sample (P=0.000005). Multiplex families, in which more than one child has autism, exhibited the strongest allelic association (P=0.000007). In case-control analyses, the relative risk of the ‘CC’ genotype was 2.26 (95% confidence interval: 1.41, 3.63) compared to the ‘GG’ genotype. Functional assays showed that the ‘C’ allele results in a 2-fold decrease in MET promoter activity and altered binding of specific transcription factor complexes. These data implicate altered MET gene expression in autism susceptibility, providing evidence of a novel pathophysiological basis for this behaviorally and medically complex disorder.

2006 - Association of a functional deficit of the BKCa channel, a synaptic regulator of neuronal excitability, with autism and mental retardation [Articolo su rivista]
Laumonnier, Frédéric; Roger, Sébastien; Guérin, Pascaline; Molinari, Florence; M'rad, Ridha; Cahard, Dominique; Belhadj, Ahlem; Halayem, Mohamed; Persico, Antonio M; Elia, Maurizio; Romano, Valentino; Holbert, Sébastien; Andres, Christian; Chaabouni, Habiba; Colleaux, Laurence; Constant, Jacques; Le Guennec, Jean-Yves; Briault, Sylvain
abstract

Autism is a complex, largely genetic psychiatric disorder. In the majority of cases, the cause of autism is not known, but there is strong evidence for a genetic etiology. To identify candidate genes, the physical mapping of balanced chromosomal aberrations is a powerful strategy, since several genes have been characterized in numerous disorders. In this study, the authors analyzed a balanced reciprocal translocation arising de novo in a subject with autism and mental retardation.

2006 - Multiple receptors mediate the trophic effects of serotonin on ventroposterior thalamic neurons in vitro [Articolo su rivista]
Persico, Antonio M.; Di Pino, Giovanni; Levitt, Pat
abstract

Serotonin (5-HT) exerts prominent morphogenetic roles during development. For example, somatosensory cortical barrel formation is altered in mouse models characterized by excessive extracellular 5-HT, suggesting that 5-HT affects development of thalamic afferents and/or neocortical target regions. The present study assessed 5-HT effects in primary cultures of fetal ventroposterior thalamic (VPT) neurons. 5-HT produces concentration-dependent trophic effects, with impressive 59% and 106% peak increases in total neurite length and number of branching points, respectively, at a dose of 30 microM 5-HT. The exposure of VPT neurons to specific 5-HT receptor agonists 8-OH-DPAT (5-HT(1A)), CGS-12066A (5-HT(1B)), DOI (5-HT(2A/2C)), and m-CPBG (5-HT(3)), enhances primary neurite length and number of branching points with rank-order potency 5-HT(1B) > 5-HT(2A/2C) = 5-HT(3) > 5-HT(1A) = vehicle. Trophic 5-HT effects on embryonic VPT neurons are thus much more prominent than previously reported, and can be mediated by multiple 5-HT receptor subtypes.

2006 - Polymorphic GGC repeat differentially regulates human reelin gene expression levels [Articolo su rivista]
Persico, A. M.; Levitt, P.; Pimenta, A. F.
abstract

The human gene encoding Reelin (RELN), a pivotal protein in neurodevelopment, includes a polymorphic GGC repeat in its 5' untranslated region (UTR). CHO cells transfected with constructs encompassing the RELN 5'UTR with 4-to-13 GGC repeats upstream of the luciferase reporter gene show declining luciferase activity with increasing GGC repeat number (P < 0.005), as predicted by computer-based simulations. Conversely, RELN 5'UTR sequences boost reporter gene expression above control levels in neuronal SN56 and N2A cell lines, but 12- and 13-repeat alleles still yield 50-60% less luciferase activity compared to the more common 8- and 10-repeat alleles (P < 0.0001). RELN "long" GGC alleles significantly blunt gene expression and may, through this effect, confer vulnerability to human disorders, such as schizophrenia and autism.

2006 - Searching for ways out of the autism maze: genetic, epigenetic and environmental clues [Articolo su rivista]
Persico, Antonio M.; Bourgeron, Thomas
abstract

Our understanding of human disorders that affect higher cognitive functions has greatly advanced in recent decades, and over 20 genes associated with non-syndromic mental retardation have been identified during the past 15 years. However, proteins encoded by "cognition genes" have such diverse neurodevelopmental functions that delineating specific pathogenetic pathways still poses a tremendous challenge. In this review, we summarize genetic, epigenetic and environmental contributions to neurodevelopmental alterations that either cause or confer vulnerability to autism, a disease primarily affecting social cognition. Taken together, these results begin to provide a unifying view of complex pathogenetic pathways that are likely to lead to autism spectrum disorders through altered neurite morphology, synaptogenesis and cell migration. This review is part of the INMED/TINS special issue "Nature and nurture in brain development and neurological disorders", based on presentations at the annual INMED/TINS symposium (http://inmednet.com/).

2005 - Clinical evidence of decreased olfaction in Bardet-Biedl syndrome caused by a deletion in the BBS4 gene [Articolo su rivista]
Iannaccone, Alessandro; Mykytyn, Kirk; Persico, Antonio M.; Searby, Charles C.; Baldi, Alfonso; Jablonski, Monica M.; Sheffield, Val C.
abstract

Recent discoveries have lead to the hypothesis that ciliary dysfunction is a mechanism underlying the pathogenesis of Bardet-Biedl syndrome (BBS). Here, we describe two individuals with decreased olfaction who are members of an extended family affected with BBS caused by a homozygous deletion (c.77-220del) in the BBS4 gene. These findings correlate with the evidence that several BBS proteins, including BBS4, are expressed in the olfactory epithelium (OE). Although the prevalence and the spectrum of impaired olfaction in BBS are not known, the causal relationship of the BBS4 deletion in this family and the decreased olfaction is corroborated by evidence that Bbs2 and Bbs4 knockout mice have severe olfaction deficits and that also patients with BBS caused by mutations in other BBS genes can have impaired olfaction. This finding broadens the spectrum of clinical manifestations associated with BBS, confirms the role of BBS4 in olfaction, and lends support to the hypothesis that ciliary dysfunction is an important aspect of BBS pathogenesis.

2005 - Paraoxonase gene variants are associated with autism in North America, but not in Italy: Possible regional specificity in gene-environment interactions [Articolo su rivista]
D'Amelio, M.; Ricci, I.; Sacco, R.; Liu, X.; D'Agruma, L.; Muscarella, L. A.; Guarnieri, V.; Militerni, R.; Bravaccio, C.; Elia, M.; Schneider, C.; Melmed, R.; Trillo, S.; Pascucci, T.; Puglisi-Allegra, S.; Reichelt, K. -L.; Macciardi, F.; Holden, J. J. A.; Persico, A. M.
abstract

Organophosphates (OPs) are routinely used as pesticides in agriculture and as insecticides within the household. Our prior work on Reelin and APOE delineated a gene-environment interactive model of autism pathogenesis, whereby genetically vulnerable individuals prenatally exposed to OPs during critical periods in neurodevelopment could undergo altered neuronal migration, resulting in an autistic syndrome. Since household use of OPs is far greater in the USA than in Italy, this model was predicted to hold validity in North America, but not in Europe. Here, we indirectly test this hypothesis by assessing linkage/association between autism and variants of the paraoxonase gene (PON1) encoding paraoxonase, the enzyme responsible for OP detoxification. Three functional single nucleotide polymorphisms, PON1 C-108T, L55M, and Q192R, were assessed in 177 Italian and 107 Caucasian-American complete trios with primary autistic probands. As predicted, Caucasian-American and not Italian families display a significant association between autism and PON1 variants less active in vitro on the OP diazinon (R192), according to case-control contrasts (Q192R: chi2=6.33, 1 df, P<0.025), transmission/disequilibrium tests (Q192R: TDT chi2=5.26, 1 df, P<0.025), family-based association tests (Q192R and L55M: FBAT Z=2.291 and 2.435 respectively, P<0.025), and haplotype-based association tests (L55/R192: HBAT Z=2.430, P<0.025). These results are consistent with our model and provide further support for the hypothesis that concurrent genetic vulnerability and environmental OP exposure may possibly contribute to autism pathogenesis in a sizable subgroup of North American individuals.

2004 - A HOX gene mutation in a family with isolated congenital vertical talus and Charcot-Marie Tooth disease [Articolo su rivista]
Shrimpton, Antony E.; Levinsohn, E. Mark; Yozawitz, Justin M.; Packard Jr., David S.; Cady, Robert B.; Middleton, Frank A.; Persico, Antonio M.; Hootnick, David R.
abstract

Congenital vertical talus (CVT), also known as “rocker-bottom foot” deformity, is a dislocation of the talonavicular joint, with rigid dorsal dislocation of the navicular over the neck of the talus. This condition is usually associated with multiple other congenital deformities and only rarely is an isolated deformity. The reported familial cases are consistent with an autosomal dominant mode of inheritance with incomplete penetrance. In contrast, Charcot-Marie- Tooth disease (CMT) is thought to be a completely distinct heterogeneous group of disorders, with foot abnormalities that typically develop a high-arched “claw foot” appearance later in life. In the present study, DNA was isolated from 36 members of a single upstate (northern) New York white family of Italian descent in which both CVT and CMT were segregating. Whole-genome linkage analysis with Affymetrix GeneChip Mapping 10K Array defined a 7-Mb critical region on chromosome 2q31, which led to candidate-gene sequencing of six HOX genes and detection of a single missense mutation, M319K (956TrA), in the HOXD10 gene. In the study family, this mutation was fully penetrant and exhibited significant evidence of linkage (LOD 6.33; v p 0), and it very likely accounts for both CVT and CMT in heterozygotes.

2004 - Association between the HOXA1 A218G polymorphism and increased head circumference in patients with autism [Articolo su rivista]
Conciatori, Monica; Stodgell, Christopher J.; Hyman, Susan L.; O'Bara, Melanie; Militerni, Roberto; Bravaccio, Carmela; Trillo, Simona; Montecchi, Francesco; Schneider, Cindy; Melmed, Raun; Elia, Maurizio; Crawford, Lori; Spence, Sarah J.; Muscarella, Lucianna; Guarnieri, Vito; D'Agruma, Leonardo; Quattrone, Alessandro; Zelante, Leopoldo; Rabinowitz, Daniel; Pascucci, Tiziana; Puglisi-Allegra, Stefano; Reichelt, Karl-Ludvig; Rodier, Patricia M.; Persico, Antonio M.
abstract

Background: The HOXA1 gene plays a major role in brainstem and cranial morphogenesis. The G allele of the HOXA1 A218G polymorphism has been previously found associated with autism (Ingram et al. 2000a). Methods: We performed case-control and family-based association analyses, contrasting 127 autistic patients with 174 ethnically-matched controls, and assessing for allelic transmission disequilibrium in 189 complete trios. Results: A, and not G alleles, were associated with autism using both case-control (2=8.96 and 5.71, 1df, P<0.005 and <0.025 for genotypes and alleles, respectively), and family-based (TDT2=8.80, 1df, P<0.005) association analyses. The head circumference of 31 patients carrying one or two copies of the G allele displayed significantly larger median values (95.0th vs 82.5th percentile, P<0.05) and dramatically reduced interindividual variability (P<0.0001), compared to 166 patients carrying the A/A genotype. Conclusions: the HOXA1 A218G polymorphism explains approximately 5% of the variance in the head circumference of autistic patients, and represents to our knowledge the first known gene variant providing sizable contributions to cranial morphology. The disease specificity of this finding is currently being investigated. Non-replications in genetic linkage/association studies could partly stem from the dyshomogeneous distribution of an endophenotype morphologically defined by cranial circumference.

2004 - Enhanced APOE2 transmission rates in families with autistic probands [Articolo su rivista]
Persico, A M; D'Agruma, L; Zelante, L; Militerni, R; Bravaccio, C; Schneider, C; Melmed, R; Trillo, S; Montecchi, F; Elia, M; Palermo, M; Rabinowitz, D; Pascucci, T; Puglisi-Allegra, S; Reichelt, K-L; Muscarella, L; Guarnieri, V; Melgari, J-M; Conciatori, M; Keller, F
abstract

We have previously described linkage/association between reelin gene polymorphisms and autistic disorder. APOE also participates in the Reelin signaling pathway, by competitively antagonizing Reelin binding to APOE receptor 2 and to very-low-density lipoprotein receptors. The APOE2 protein variant displays the lowest receptor binding affinity compared with APOE3 and APOE4. In this study, we assess linkage/association between primary autism and APOE alleles in 223 complete trios, from 119 simplex Italian families and 44 simplex and 29 multiplex Caucasian-American families. Statistically significant disequilibrium favors the transmission of epsilon2 alleles to autistic offspring, over epsilon3 and epsilon4 (allele-wise transmission/disequilibrium test [TDT], chi2 = 6.16, 2 degrees of freedom [d.f.], P<0.05; genotype-wise TDT, chi2 = 10.68, 3 d.f., P<0.05). A novel epsilon3r allele was also discovered in an autistic child and his mother. Autistic patients do not differ significantly from unaffected siblings (allele-wise TDT comparing autistic patients versus unaffected sibs, chi2 = 1.83, 2 d.f., P<0.40, not significant). The major limitation of this study consists of our small sample size of trios including one unaffected sibling, currently not possessing the statistical power necessary to conclusively discriminate a specific association of epsilon2 with autism, from a distorted segregation pattern characterized by enhanced epsilon2 transmission rates both to affected and unaffected offspring. Our findings are thus compatible with either (a) pathogenetic contributions by epsilon2 alleles to autism spectrum vulnerability, requiring additional environmental and/or genetic factors to yield an autistic syndrome, and/or (b) a protective effect of epsilon2 alleles against the enhanced risk of miscarriage and infertility previously described among parents of autistic children.

2004 - Roles for serotonin in neurodevelopment: more than just neural transmission [Articolo su rivista]
Di Pino, G.; Moessner, R.; Lesch, K. P.; Lauder, J. M.; Persico, A. M.
abstract

During pre- and early postnatal development, the neurotransmitter serotonin (5-HT) modulates cell proliferation, migration and programmed cell death, as well as cell shape and cell-cell coupling. These “trophic” effects of 5-HT, involving the cytoskeletal function, the cell cycle, and programmed cell death, can be both dependent and independent of the changes in resting membrane potential that typically define neurotransmitter action. The morphogenetic role of 5-HT is neither limited to the central nervous system (CNS), nor does it impinge upon just a single aspect of cell biology. Ontogenic differences in regional and temporal expression patterns of 5-HT receptors mediating these effects in different systems add further complexity. This review summarizes neurobiological evidence for the trophic involvement of 5-HT during development and discusses related medical issues, including potential teratological risks and possible novel therapeutic indications of selective serotonin reuptake inhibitor (SSRI) administration.

2003 - Methodological factors influencing measurement and processing of plasma reelin in humans [Articolo su rivista]
Lugli, Giovanni; Krueger, Jacqueline M.; Davis, John M.; Persico, Antonio M.; Keller, Flavio; Smalheiser, Neil R.
abstract

Background: Reelin, intensively studied as an extracellular protein that regulates brain development, is also expressed in a variety of tissues and a circulating pool of reelin exists in adult mammals. Here we describe the methodological and biological foundation for carrying out and interpreting clinical studies of plasma reelin. Results: Reelin in human plasma was sensitive to proteolysis, freeze-thawing and heating during long-term storage, sample preparation and electrophoresis. Reelin in plasma was a dimer under denaturing conditions. Boiling of samples resulted in laddering, suggesting that each of the 8 repeats expressed in reelin contains a heat-labile covalent bond susceptible to breakage. Urinary-type and tissue-type plasminogen activator converted reelin to a discrete 310 kDa fragment co-migrating with the major immunoreactive reelin fragment seen in plasma and also detected in brain. (In contrast, plasmin produced a spectrum of smaller unstable reelin fragments.) We examined archival plasma of 10 pairs of age-matched male individuals differing in repeat length of a CGG repeat polymorphism of the 5'-untranslated region of the reelin gene (both alleles &lt; 11 repeats vs. one allele having &gt;11 repeats). Reelin 310 kDa band content was lower in subjects having the long repeats in all 10 pairs, by 25% on average (p &lt; 0.001). In contrast, no difference was noted for amyloid precursor protein. Conclusions: Our studies indicate the need for caution in measuring reelin in archival blood samples, and suggest that assays of plasma reelin should take into account three dimensions that might vary independently: a) the total amount of reelin protein; b) the relative amounts of reelin vs. its proteolytic processing products; and c) the aggregation state of the native protein. Reelin plasminogen activator interactions may affect their roles in synaptic plasticity. Our results also suggest that the human CGG repeat polymorphism affects reelin gene expression, and may affect susceptibility to human disease.

2003 - Reduced programmed cell death in brains of serotonin transporter knockout mice [Articolo su rivista]
Persico, Antonio M; Baldi, Alfonso; Dell'Acqua, Maria Luisa; Moessner, Rainald; Murphy, Dennis L; Lesch, Klaus-Peter; Keller, Flavio
abstract

Serotonin (5-HT) is known to reduce apoptosis and in rodent models of brain ischemia. Modulation of programmed cell death during neural development was assessed in early postnatal brains of serotonin transporter (5-HTT) knockout mice, characterized by elevated extracellular 5-HT levels. The number of apoptotic cells visualized at postnatal day-1 (P1) by ISEL+ or TUNEL staining was significantly reduced in the striatum, thalamus/hypothalamus, cerebral cortex and hippocampus of 5-HTT knockout mice, compared to wild type and heterozygote mice, with differences displaying an increasing fronto-caudal gradient and regional specificity. These findings underscore 5-HT roles in the regulation of programmed cell death during brain development, and spur interest into pharmacological interventions aimed at relieving pathological apoptosis by potentiating serotoninergic neurotransmission.

2003 - Serotonergic regulation of somatosensory cortical development: Lessons from genetic mouse models [Articolo su rivista]
Luo, Xiaoyan; Persico, Antonio M.; Lauder, Jean M.
abstract

Monoaminergic neurotransmitter systems appear early during embryogenesis, suggesting that they could play important roles in brain development. Accumulated evidence indicates that serotonin (5-hydroxytryptamine, 5-HT) regulates neural as well as nonneural development, including early aspects of embryonic development, differentiation of neuronal progenitors, and morphogenesis of the craniofacial region, heart and limb. Recent studies using monoamine oxidase-A (MAO-A), 5-HT transporter, vesicular monoamine transporter-2 (VMAT2) and 5-HT1B receptor single, double and triple knockout mice have provided evidence that the serotonergic system plays important roles in barrel field formation in the developing somatosensory cortex. Here we review evidence from these genetic mouse models and, based on the accumulated evidence, propose a testable model for future studies of mechanisms underlying serotonergic regulation of cortical development.

2003 - Subjective tinnitus, temporomandibular joint dysfunction, and serotonin modulation of neural plasticity: causal or casual triad? [Articolo su rivista]
Salvinelli, F.; Casale, M.; Paparo, F.; Persico, A. M.; Zini, C.
abstract

Tinnitus and temporomandibular joint dysfunction (TMJD) are among the most common complaints encountered by physicians. Though the relationship between tinnitus and TMJD has attracted great interest during the past several years, theories attempting to explain this association are still few and inconsistent. Conceivably, TMJD could irritate auricolo-temporal nerve (ATN), triggering a somatosensory pathway-induced disinhibition of dorsal cochlear nucleus (DCN) activity in the auditory pathway. In genetically-predisposed TMJD patients, signals from cronically stimulated DCNs activating specific cortical neuronal networks, could yield plastic neural changes resulting in tinnitus. Based on current evidence of serotoninergic modulation of neural activity and plasticity in sensory pathways, reduced serotoninergic tone could promote plastic changes underlying tinnitus through diminished filtering of incoming signals. Therefore, the early establishment of specific treatments aimed at improving TMJD and/or boosting serotoninergic activity may be required to prevent the creation of 'tinnitus memory circuits'.

2003 - The Neurobiological Context of Autism [Articolo su rivista]
Keller, Flavio; Persico, Antonio M.
abstract

Autistic disorder (AD) is a complex neuropsychiatric disorder of neurodevelopmental origin, where multiple genetic and environmental factors may interact, resulting in a clinical continuum. The genetic component is best described by a multilocus model that takes into account epistatic interactions between several susceptibility genes. In the past ten years enormous progress has been made in identifying chromosomal regions in linkage with AD, but no single gene has emerged yet as a major factor of genetic liability. Neuroanatomical findings point to early dysgenetic events taking place in the cerebral cortex, cerebellum, and brains stem. At the cellular level, disease mechanisms may include altered cell migration, increased cell proliferation, decreased cell death, or altered synapse elimination. In contrast to other diseases of the nervous system (e.g. Parkinson’s and Alzheimer’s disease) neurochemical findings in AD point to involvement of multiple neurotransmitter systems. The serotoninergic system has been intensively investigated in this respect, but other neurotrasmitter systems known to be involved in cognitive functions (e.g. the cholinergic system) are coming under closer scrutiny. The role of environmental factors is also coming under closer investigation. It is not clear yet whether environmental factors act merely as precipitating agents, always requiring an underlying genetic liability, or whether they represent an essential component of a pathogenetic process where genetic liability alone does not lead to the full-blown autism phenotype.

2002 - Erratum: Reelin is a serine protease of the extracellular matrix (Journal of Biological Chemistry (2002) 277 (303-309)) [Articolo su rivista]
Quattrocchi, C. C.; Wannenes, F.; Persico, Antonio; Ciafre, S. A.; D'Arcangelo, G.; Farace, M. G.; Keller, F.
abstract

2002 - Increased hippocampal DNA oxidation in serotonin transporter deficient mice [Articolo su rivista]
Mössner, R.; Dringen, R.; Persico, A. M.; Janetzky, B.; Okladnova, O.; Albert, D.; Götz, M.; Benninghoff, J.; Schmitt, A.; Gerlach, M.; Riederer, P.; Lesch, K. P.
abstract

The serotonin transporter (5HTT) is the molecule responsible for the high-affinity reuptake of 5HT from the synaptic cleft. Mice lacking the 5HTT exhibit highly elevated extracellular concentrations of 5HT. We assessed whether the glutathione detoxification system is altered in 5HTTdeficient mice. While levels of reduced and oxidized glutathione were unchanged, glutathione metabolising enzymes showed a differential pattern of modulation. Glutathione peroxidase was reduced in frontal cortex, brainstem, and cerebellum of 5HTT-deficient mice, though not to a statistically significant extent, while a putative isoform of the detoxifying enzyme glutathione-S-transferase pi was decreased in a number of brain regions, especially in brainstem. At the level of the DNA, we found an increase of oxidative DNA adducts in the hippocampus of 5HTT-deficient mice. Given the importance of the hippocampus in learning and memory, this may be the most important neurochemical consequence of the absence of the 5HTT.

2002 - Reelin is a serine protease of the extracellular matrix [Articolo su rivista]
Quattrocchi, Carlo C.; Wannenes, Francesca; Persico, Antonio M.; Ciafré, Silvia Anna; D'Arcangelo, Gabriella; Farace, Maria G.; Keller, Flavio
abstract

Reelin is an extracellular matrix protein that plays a pivotal role in development of the central nervous system. Reelin is also expressed in the adult brain, notably in the cerebral cortex, where it might play a role in synaptic plasticity. The mechanism of action of reelin at the molecular level has been the subject of several hypotheses. Here we show that reelin is a serine protease and that proteolytic activity is relevant to its function, since (i) Reelin expression in HEK 293T cells impairs their ability to adhere to fibronectin-coated surfaces, and adhesion to fibronectin is restored by micromolar concentrations of diisopropyl phosphorofluoridate, a serine hydrolase inhibitor; (ii) purified Reelin binds FPPeg-biotin, a trap probe which irreversibly binds to serine residues located in active catalytic sites of serine hydrolases; (iii) purified Reelin rapidly degrades fibronectin and laminin, while collagen IV is degraded at a much slower rate; fibronectin degradation is inhibited by inhibitors of serine proteases, and by monoclonal antibody CR-50, an antibody known to block the function of Reelin both in vitro and in vivo. The proteolytic activity of Reelin on adhesion molecules of the extracellular matrix and/or receptors on neurons may explain how Reelin regulates neuronal migration and synaptic plasticity.

2002 - Serotonin transporter promoter variants do not explain the hyperserotoninemia in autistic children [Articolo su rivista]
Persico, A. M.; Pascucci, T.; Puglisi-Allegra, S.; Militerni, R.; Bravaccio, C.; Schneider, C.; Melmed, R.; Trillo, S.; Montecchi, F.; Palermo, M.; Rabinowitz, D.; Reichelt, K. -L.; Conciatori, M.; Marino, R.; Keller, F.
abstract

Autism is a biologically-heterogeneous disease. Distinct subgroups of autistic patients may be marked by intermediate phenotypes, such as elevated serotonin (5-HT) blood levels, potentially associated with different underlying disease mechanisms. This could lead to inconsistent genetic association results, such as those of prior studies on serotonin transporter (5-HTT) gene promoter variants and autistic disorder. Contributions of 5-HTT gene promoter alleles to 5-HT blood levels were thus investigated in 134 autistic patients and 291 first-degree relatives. Mean 5-HT blood levels are 11% higher in autistic patients carrying the L/L genotype, compared to patients with the S/S or S/L genotype; this trend is not observed in first-degree relatives. The probability of inheriting L or S alleles is significantly enhanced in patients with 5-HT blood levels above or below the mean, respectively (P 0.05), but quantitative TDT analyses yield a non-significant trend (P = 0.10), as this polymorphism explains only 2.5% of the variance in 5-HT blood levels of autistic patients. In conclusion, 5-HTT gene promoter variants seemingly exert a small effect on 5-HT blood levels in autistic children, which largely does not account for hyperserotoninemia. Nonetheless, the inconsistent outcome of prior association studies could partly stem from a selection bias of hyper- or hypo-serotoninemic probands.

2001 - Barrel pattern formation requires serotonin uptake by thalamocortical afferents, and not vesicular monoamine release [Articolo su rivista]
Persico, A M; Mengual, E; Moessner, R; Hall, F S; Revay, R S; Sora, I; Arellano, J; Defelipe, J; Gimenez-Amaya, J M; Conciatori, M; Marino, R; Baldi, A; Cabib, S; Pascucci, T; Uhl, G R; Murphy, D L; Lesch, K P; Keller, F; Hall, S F
abstract

Thalamocortical neurons innervating the barrel cortex in neonatal rodents transiently store serotonin (5-HT) in synaptic vesicles by expressing the plasma membrane serotonin transporter (5-HTT) and the vesicular monoamine transporter (VMAT2). 5-HTT knock-out (ko) mice reveal a nearly complete absence of 5-HT in the cerebral cortex by immunohistochemistry, and of barrels, both at P7 and adulthood. Quantitative electron microscopy reveals that 5-HTT ko affects neither the density of synapses nor the length of synaptic contacts in layer IV. VMAT2 ko mice, completely lacking activity-dependent vesicular release of monoamines including 5-HT, also show a complete lack of 5-HT in the cortex but display largely normal barrel fields, despite sometimes markedly reduced postnatal growth. Transient 5-HTT expression is thus required for barrel pattern formation, whereas activity-dependent vesicular 5-HT release is not.

2001 - No association between the 4g/5G polymorphism of the plasminogen activator inhibitor-1 gene promoter and autistic disorder [Articolo su rivista]
Persico, A M; Militerni, R; Bravaccio, C; Schneider, C; Melmed, R; Trillo, S; Montecchi, F; Palermo, M; Pascucci, T; Puglisi-Allegra, S; Reichelt, K L; Conciatori, M; Keller, F
abstract

Plasmin, a serine protease, is involved in many physiologically relevant processes, including haemostasis, cellular recruitment during immune response, tumour growth, and also neuronal migration and synaptic remodelling. Both tissue-type and urokinase-type plasminogen activators can be efficiently inhibited by plasminogen activator inhibitor-1 (PAI-1), a protease inhibitor of the serpin family. The human PAI-1 gene is located on chromosome 7q, within or close to a region that has been linked to autism in several linkage studies. Autism seems to be characterized by altered neuronal cytoarchitecture, synaptogenesis and possibly also cellular immune responses. We began addressing the potential involvement of the PAI-1 gene in autistic disorder with this linkage/association study, assessing transmission patterns of the 4G/5G polymorphism in the PAI-1 gene promoter that was previously shown to significantly affect PAI-1 plasma levels. No linkage/association was found in 167 trios with autistic probands, recruited in Italy and in the USA. We thus found no evidence that this polymorphism, or putative functionally relevant gene variants in linkage disequilibrium with it, confer vulnerability to autistic disorder.

2001 - Reelin gene alleles and haplotypes as a factor predisposing to autistic disorder [Articolo su rivista]
Persico, A. M.; D'agruma, L.; Maiorano, N.; Totaro, A.; Militerni, R.; Bravaccio, C.; Wassink, T. H.; Schneider, C.; Melmed, R.; Trillo, S.; Montecchi, F.; Palermo, M.; Pascucci, T.; Puglisi-Allegra, S.; Reichelt, K. -L.; Conciatori, M.; Marino, R.; Quattrocchi, C. C.; Baldi, A.; Zelante, L.; Gasparini, P.; Keller, F.
abstract

Autistic disorder (MIM 209850) is currently viewed as a neurodevelopmental disease. Reelin plays a pivotal role in the development of laminar structures including the cerebral cortex, hippocampus, cerebellum and of several brainstem nuclei. Neuroanatomical evidence is consistent with Reelin involvement in autistic disorder. In this study, we describe several polymorphisms identified using RNA-SSCP and DNA sequencing. Association and linkage were assessed comparing 95 Italian patients to 186 ethnically-matched controls, and using the transmission/disequilibrium test and haplotype-based haplotype relative risk in 172 complete trios from 165 families collected in Italy and in the USA. Both case-control and family-based analyses yield a significant association between autistic disorder and a polymorphic GGC repeat located immediately 59 of the reelin gene (RELN) ATG initiator codon, as well as with specific haplotypes formed by this polymorphism with two single-base substitutions located in a splice junction in exon 6 and within exon 50. Triplet repeats located in 59 untranslated regions (59UTRs) are indicative of strong transcriptional regulation. Our findings suggest that longer triplet repeats in the 59UTR of the RELN gene confer vulnerability to autistic disorder.

2000 - Adenosine Deaminase (ADA) alleles and autistic disorder: case-control and family-based association studies [Articolo su rivista]
Persico, A. M.; Militerni, R.; Bravaccio, C.; Schneider, C.; Melmed, R.; Trillo, S.; Montecchi, F.; Palermo, M.; Pascucci, T.; PUGLISI-ALLEGRA, S.; Reichelt, K. -L.; Conciatori, M.; Baldi, A.; Keller, F.
abstract

2000 - Adenosine deaminase alleles and autistic disorder: case-control and family-based association studies [Articolo su rivista]
Persico, A M; Militerni, R; Bravaccio, C; Schneider, C; Melmed, R; Trillo, S; Montecchi, F; Palermo, M T; Pascucci, T; Puglisi-Allegra, S; Reichelt, K L; Conciatori, M; Baldi, A; Keller, F
abstract

Adenosine deaminase (ADA) plays a relevant role in purine metabolism, immune responses, and peptidase activity, which may be altered in some autistic patients. Codominant ADA1 and ADA2 alleles code for ADA1 and ADA2 allozymes, the most frequent protein isoforms in the general population. Individuals carrying one copy of the ADA2 allele display 15 to 20% lower catalytic activity compared to ADA1 homozygotes. Recent preliminary data suggest that ADA2 alleles may be more frequent among autistic patients than healthy controls. The present study was undertaken to replicate these findings in a new case-control study, to test for linkage/association using a family-based design, and to characterize ADA2-carrying patients by serotonin blood levels, peptiduria, and head circumference. ADA2 alleles were significantly more frequent in 91 Caucasian autistic patients of Italian descent than in 152 unaffected controls (17.6% vs. 7.9%, P = 0.018), as well as among their fathers. Family-based tests involving these 91 singleton families, as well as 44 additional Caucasian-American trios, did not support significant linkage/association. However, the observed preferential maternal transmission of ADA2 alleles, if replicated, may point toward linkage disequilibrium between the ADA2 polymorphism and an imprinted gene variant located in its vicinity. Racial and ethnic differences in ADA allelic distributions, together with the low frequency of the ADA2 allele, may pose methodological problems to future linkage/association studies. Direct assessments of ADA catalytic activity in autistic individuals and unaffected siblings carrying ADA1/ADA1 vs ADA1/ADA2 genotypes may provide stronger evidence of ADA2 contributions to autistic disorder. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:784-790, 2000.

2000 - Deafferentation-induced apoptosis of neurons in thalamic somatosensory nuclei of the newborn rat: critical period and rescue from cell death by peripherally applied neurotrophins [Articolo su rivista]
A., Baldi; E., Calia; A., Ciampini; Riccio, Massimo; A., Vetuschi; A. M., Persico; F., Keller
abstract

This study shows that unilateral transection of the infraorbital nerve (ION) in newborn (P0) rats induces apoptosis in the contralateral ventrobasal thalamic (VB) complex, as evidenced by terminal transferase-mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) and electron miscroscopy. Double-labelling experiments using retrograde transport of labelled microspheres injected into the barrel cortex, followed by TUNEL staining, show that TUNEL-positive cells are thalamocortical neurons. The number of TUNEL-positive cells had begun to increase by 24 h postlesion, increased further 48 h after nerve section, and decreased to control levels after 120 h. Lesion-induced apoptosis in the VB complex is less pronounced if ION section is performed at P4, and disappears if the lesion is performed at P7. This time course closely matches the critical period of lesion-induced plasticity in the barrel cortex. Nerve growth factor (NGF) or brain-derived neurotrophic factor (BDNF), applied on the ION stump alone or in combination, are able to partially rescue thalamic neurons from apoptosis. Total cell counts in the VB complex of P7 animals that underwent ION section at P0 confirm the rescuing effect of BDNF and NGF. Blockade of axonal transport in the ION mimics the effect of ION section. These data suggest that survival-promoting signals from the periphery, maybe neurotrophins, are required for the survival of higher-order neurons in the somatosensory system during the period of fine-tuning of neuronal connections. We also propose that anterograde transneuronal degeneration in the neonatal rat trigeminal system may represent a new animal model for studying the pathways of programmed cell death in vivo.

2000 - Differential regulation of adenosine A1 and A(2A) receptors in serotonin transporter and monoamine oxidase A-deficient mice [Articolo su rivista]
Mössner, Rainald; Albert, Dietmar; Persico, Antonio M.; Hennig, Thomas; Bengel, Dietmar; Holtmann, Bettina; Schmitt, Angelika; Keller, Flavio; Simantov, Rabi; Murphy, Dennis; Seif, Isabelle; Deckert, Jürgen; Lesch, K. Peter
abstract

The serotonin (5HT) transporter (5HTT) removes 5HT from the synaptic cleft and is thus critical to the control of serotonergic neurotransmission. Mice with a targeted inactivation of the 5HTT represent a novel and unique tool to study serotonergic system functioning. Because the release of 5HT is regulated by adenosine, we investigated 5HTT-deficient mice for possible adaptive changes of adenosine A and A receptors. A and A receptors were studied by means of quantitative autoradiography using the radioligands 1 2A 1 2A 3 3 [ H]8-cyclopentyl-1,3-dipropylxanthine and [ H]CGS 21680, respectively. A comparison of 5HTT knockout versus control mice revealed upregulation of A receptors in the dorsal raphe nucleus (DRN, 121%), but not in any of the serotonergic projection areas, and 1 downregulation of A receptors in basal ganglia. The adaptive changes of A and A receptors in 5HTT-deficient mice are likely to 2A 1 2A represent a compensatory neuroprotective effect mediated by the adenosinergic modulatory system. For comparison, these receptors were also studied in monoamine oxidase A (MAOA) knockout mice and in 5HTT/MAOA double knockout mice. 5HTT/MAOA double knockout mice showed adaptive changes of adenosine A and A receptors similar to 5HTT knockout mice, while investigation of 1 2A MAOA-deficient mice revealed an upregulation of A receptors, which may relate to a role of both MAOA and adenosine A receptors 2A 2A in anxiety.

2000 - Lack of association between serotonin transporter gene promoter variants and autistic disorder in two ethnically-distinct samples [Articolo su rivista]
Persico, A. M.; Militerni, R.; Bravaccio, C.; Schneider, C.; Melmed, R.; Damiani, V.; Baldi, A.; Keller, F.
abstract

Family-based studies performed to date provide conflicting evidence of linkage/association between autistic disorder and either the "short" [Cook et al., 1997: Mol Psychiatry 2:247-250] or the "long" [Klauck et al., 1997: Hum Mol Genet 6:2233-2238] allele of a polymorphic repeat located in the serotonin transporter (5-HTT) gene promoter region, affecting 5-HTT gene expression [Lesch et al., 1996: Science 274:1527-1531]. The present study was designed to assess linkage and linkage disequilibrium in two new ethnically distinct samples of families with primary autistic probands. The 5-HTT promoter repeat was genotyped in 54 singleton families collected in Italy and in 32 singleton and 5 multiplex families collected in the U.S.A., yielding a total sample of 98 trios. Linkage/association between 5-HTT gene promoter alleles and autistic disorder was assessed using the transmission/disequilibrium test (TDT) and the haplotype-based haplotype relative risk (HHRR). Both the Italian and the American samples, either singly or combined, displayed no evidence of linkage/association between 5-HTT gene promoter alleles and autistic disorder. Our findings do not support prominent contributions of 5-HTT gene variants to the pathogenesis of idiopathic infantile autism. Heterogeneity in pathogenetic mechanisms underlying the disease may require that linkage/association studies be targeted toward patient subgroups isolated on the basis of specific biochemical markers, such as serotonin (5-HT) blood levels. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:123-127, 2000.

2000 - Serotonin depletion and barrel cortex development: impact of growth impairment vs. serotonin effects on thalamocortical endings [Articolo su rivista]
Persico, A M; Altamura, C; Calia, E; Puglisi-Allegra, S; Ventura, R; Lucchese, F; Keller, F
abstract

Converging evidence supports a role of serotonin (5-hydroxytryptamine; 5-HT) in barrel cortex development. Systemic administration of 5-HT-depleting drugs reduces cross-sectional whisker barrel areas in the somatosensory cortex (SSC) of neonatal rats. Here we assess the relative impact on barrel pattern formation of (i) 5-HT depletion and (ii) decreased brain growth, which is often associated with pharmacological 5-HT depletion, by comparing the effects of 5-HT-depleting drugs with those of reduced protein intake. Left hemisphere 5-HT levels in the SSC and right hemisphere whisker barrel areas were assessed at postnatal day 6 (P6) in the same animal following injection of p-chloroamphetamine (PCA) or p-chlorophenylalanine (PCPA) at P0. Both drugs significantly reduced cortical 5-HT content and mean barrel areas at P6, but also body and brain growth. Differences in brain weight accounted for 84.4% of the variance in barrel size, with negligible contributions by cortical 5-HT content. PCPA-treated animals sacrificed at P14 yielded similar trends, albeit less pronounced. Finally, reduced protein intake resulted in lower body weight and cortical 5-HT levels at P6, but yielded no change in brain weight or mean barrel area. Barrel formation therefore appears markedly less sensitive to 5-HT depletion per se than to drug-induced growth impairment.

1999 - La genética de la adicción a drogas: de los marcadores genéticos al comportamiento humano [Articolo su rivista]
Persico, A. M.
abstract

RESUMEN: Objetivo: presentar una actualización de las evidencias que sugieren un componente genético en la etiología de los trastornos adictivos. Material y métodos: revisión crítica de los hallazgos publicados en estudios familiares y genes candidatos del sistema dopaminérgico. Resultados: los factores genéticos contribuyen a la vulnerabilidad al abuso de sustancias. Los hallazgos relativos a los genes del receptor dopamina D2 (DRD2) y su relación con el alcoholismo y el abuso de sustancias son controvertidos. No existen conclusiones firmes respecto a la relación entre los genes del receptor dopamina D4 (DRD4) y la tipología de búsqueda de sensaciones nuevas. Conclusiones: deben desarrollarse criterios para la evaluación de los estudios de asociación. El estudio de subgrupos de pacientes identificados a través de marcadores biológicos puede incrementar la homogeneidad de las muestras estudiadas.

1998 - BDNF and NT-3 applied in the whisker pad reverse cortical changes after peripheral deafferentation in neonatal rats [Articolo su rivista]
Calia, E.; Persico, A. M.; Baldi, A.; Keller, F.
abstract

It has been known for a long time that subcortical input drives the specification of cortical areas. Molecular signals mediating this instructive effect from the periphery are poorly understood. In foetal or neonatal rats, ablation of whisker follicles, transection of the infraorbital nerve, inhibition of axonal transport, but not impulse activity blockade, prevent formation of barrels in the primary somatosensory cortex (S1). These findings suggest that a chemical signal, possibly arising from the skin or the follicle, may be responsible for somatotopic pattern formation in S1. Neurotrophins promote survival and differentiation of primary sensory neurons, and are expressed in the whisker pad during development. Neonatal rats received gelfoam impregnated with NGF, BDNF or NT-3 under the whisker pad following surgical denervation of whisker rows D and E on P0. Barrel formation in S1 was assessed on P7 by acetylcholinesterase histochemistry and 5-HT-immunohistochemistry. BDNF and NT-3, but not NGF, promoted development of the cortical barrels corresponding to denervated whiskers. Furthermore, BDNF and NT-3 prevented the lesion-induced expansion of row C barrels, while NGF appeared to promote row C expansion. Our results suggest that BDNF and NT-3 arising from the whisker pad are involved in the formation and/or maintenance of the barrel pattern in S1. These findings are potentially relevant for the prevention of sensory disturbances possibly due to reorganization of central sensory circuits after peripheral nerve lesions in humans.

1998 - Lack of association between dopamine transporter gene polymorphisms and delusional disorder [Articolo su rivista]
Persico, A M; Catalano, M
abstract

Potential contributions of dopamine transporter (DAT) gene variants to delusional disorder were investigated using association analysis. DAT gene VNTR polymorphisms were assessed in 61 delusional patients and 54 normal controls. No differences were found in either genotypic or allelic distributions. These findings do not support relevant contributions of DAT gene variants to the pathogenesis of delusional disorder.

1998 - Medial prefrontal cortical injections of c-fos antisense oligonucleotides transiently lower c-Fos protein and mimic amphetamine withdrawal behaviours [Articolo su rivista]
Persico, A M; Schindler, C W; Davis, S C; Ambrosio, E; Uhl, G R
abstract

Prefrontal cerebral cortical areas display decreased expression of several transcription factor/immediate-early genes, including c-fos, during amphetamine withdrawal. Antisense strategies can help to test possible roles for this prefrontal c-fos down-regulation in the behavioural correlates of amphetamine withdrawal. Medial prefrontal cortical injections delivering 1.7 nmoles of anti c-fos oligonucleotides revealed an approximately 3 h half-life for phosphothioate and a 15 min half-life for phosphodiester oligonucleotides. Antisense phosphothioates complementary to the c-fos translational start site reduced levels of c-Fos protein, while exerting modest and variable effects on c-fos messenger RNA levels. Neither missense phosphorothioate nor antisense phosphodiester oligonucleotides significantly reduced levels of either c-fos messenger RNA or protein. Animals injected with anti c-fos phosphothioate oligonucleotides into the medial prefrontal cortex displayed marked reductions in linear locomotor activity and repetitive movements measured in a novel environment, effects not seen when missense oligonucleotides were used or when animals were accustomed to the activity monitor prior to antisense oligonucleotide injection. Behavioural changes produced by prefrontal cortical injections of c-fos antisense oligonucleotides closely mimic alterations recorded during amphetamine withdrawal. Prefrontal c-fos could thus conceivably play roles in the neurobiological underpinnings of psychostimulant withdrawal and of responses to stressors such as exposure to novel environments.

1998 - Parkinsonian patients report blunted subjective effects of methylphenidate [Articolo su rivista]
Persico, A. M.; Reich, S.; Henningfield, J. E.; Kuhar, M. J.; Uhl, G. R.
abstract

Mesolimbic-mesocortical dopamine brain circuits important for psychostimulant reward in animals are developed to greater extents in humans. Brains of patients with Parkinson's disease show depletion of ventral tegmental area mesolimbic-mesocortical neurons. The authors assessed psychostimulant responses in parkinsonian patients to test whether intact dopaminergic systems are required for subjective psychostimulant effects. Responses to placebo and 15, 20, 25, and 30 mg of methylphenidate were studied in 12 parkinsonian patients and 12 neurologically intact matched controls. Physiological and subjective mood responses were recorded using the Profile of Mood Slates, Addiction Research Center Inventory, and Visual Analog Scale. Drug-induced changes in "good" feelings and overall drug responses were attenuated in the parkinsonian patients. These results, in conjunction with animal data, provide support for dopamine hypotheses of psychostimulant reward in humans and suggest possible bases for some of the mood disturbances found in many parkinsonian patients.

1997 - Erratum: The association between dopamine D2 receptor gene variants and addiction: Facts and opinions (Alcologia European Journal of Alcohol Studies (1996) 8(3) (177-184)) [Articolo su rivista]
Persico, A.
abstract

1997 - Genotypic association between dopamine transporter gene polymorphisms and schizophrenia [Articolo su rivista]
Persico, A M; Macciardi, F
abstract

Dopamine transporter (DAT) gene variants do not appear to provide widespread contributions to the etiology of schizophrenia spectrum disorders, according to linkage studies [Persico et al., 1995: Am J Psychiatry 152:134-136]. They may, however, produce modifying effects, more readily detectable in specific subpopulations of schizophrenics through associations analyses. We therefore compared polymorphic DAT gene variable number tandem repeat (VNTR) distributions in 84 controls and 147 patients, divided according to DSM-IIIR schizophrenia type criteria. No evidence of allelic association between DAT alleles and schizophrenia or any specific schizophrenia subtype was found. Interestingly, the DAT genotype distribution among schizophrenic patients did display a statistically significant departure from the genotype distribution found in controls. Such discrepancies may represent stigmata of assortative mating or may suggest a "modifying" contribution of homozygote DAT genotypes to pathogenetic processes underlying schizophrenia.

1997 - Impiego della acetil-l-carnitina nella disintossicazione dalla nicotina [Brevetto]
Persico, A. M.; Malin, D. H.
abstract

1997 - Implants for sustained drug release over the somatosensory cortex of the newborn rat: A comparison of materials and surgical procedures [Articolo su rivista]
Persico, Antonio M.; Calia, Eustachio; Keller, Flavio
abstract

Drugs that interfere with neural transmission are an important tool to assess the role of specific neurotransmitters in the development of the nervous system. Systemic drug treatments often produce neurodevelopmental effects with questionable specificity. Furthermore, many compounds of interest do not cross the blood-brain barrier. To overcome these limitations, either elvax or gelfoam implants have been previously employed to produce sustained drug release over specific brain regions. In this paper, stereotaxic coordinates are provided for reproducible insertion of drug-delivery systems over the rat somatosensory cortex at birth (P0), prior to the appearance of the cortical barrel pattern; a novel and simpler method for preparation of elvax 40p sheets is described; a new implantation technique is provided. Furthermore, we compare the efficiency and tolerability of elvax vs gelfoam implants, showing that gelfoam, but not elvax, significantly disrupts cortical cytoarchitecture. Finally, successful destruction of serotonin-containing terminals in layer IV of the primary somatosensory cortex of the newborn rat is demonstrated by application of parachloroamphetamine-containing elvax implants.

1997 - Polymorphisms of the D2 dopamine receptor gene in polysubstance abusers [Capitolo/Saggio]
Persico, A. M.; Uhl, G. R.
abstract

1996 - Contribuciòn genetica a la neurobiologia de la vulnerabilidad a la adiccion a drogas [Capitolo/Saggio]
Persico, A. M.
abstract

1996 - D2 dopamine receptor gene TaqI A1 and B1 restriction fragment length polymorphisms: Enhanced frequencies in psychostimulant-preferring polysubstance abusers [Articolo su rivista]
Persico, Antonio M.; Bird, Geoffrey; Gabbay, Frances H.; Uhl, George R.
abstract

Several lines of evidence suggest that presence of a D 2 dopamine receptor (DRD2) gene variant marked by TaqI restriction fragment length polymorphisms (RFLPs) might contribute to vulnerability to substance abuse. Psychostimulants display the most robust enhancement of dopamine activity in mesolimbic/mesocortical circuits important for behavioral reward. The present study tests the hypothesis that a DRD2 gene variant might be more prominent in polysubstance users who preferentially use psychostimulants than in addicts with preferential opiate use or in those with no drug preference. Polysubstance users with histories of heavy daily preferential psychostimulant use more often displayed one or two copies of the TaqI A1 (27/62 = 43.5% vs 33/119 = 27.7%for controls), and B1 (20/62 = 32.3% vs 23/119 = 19.8% for controls) markers at the DRD2 locus. DRD2 gene marker distributions in abusers with more prominent opiate use, or those with no history of drug preference, were similar to control genotypes. Psychostimulant-preferring drug users also reported earlier onset of psychostimulant use. Our data are consistent with the hypothesis that DRD2 gene variants marked by these polymorphisms may work, probably in concert with other genetic and environmental factors, to enhance vulnerability to psychostimulant abuse.

1996 - Dopamine D2 receptor gene polymorphisms and vulnerability to substance abuse in African Americans [Articolo su rivista]
Berrettini, Wade H.; Persico, Antonio M.
abstract

1996 - The association between dopamine D2 receptor gene [Articolo su rivista]
Persico, A
abstract

The dopamine D2 receptor (DRD2) gene locus diplays polymorphic genetic markers. Two of them, the TaqI &lt;&gt; and &lt;&gt; Restriction Fragment Length Polymorphisms (RFLP), appear to mark a DRD2 gene variant associated with enhanced liability to severe alcoholism or substance abuse in most studies. Other reports, however, show non-significant trends or fail to repliate the association altogether. Positive findings have thus been criticized as potentially stemming from RFLP frequency differences in distinct Caucasian ethnic groups, a population-genetic phenomenon entirely unrelated to alcoholism per se. Negative results have been viewed, on the other hand, as possibly deriving from a biased selection of non-severe patients in a genetically heterogeneous disorder such as alcoholism. Data from studies published to date will be reviewed, opposing views will be discussed, meta-analyses consistent with contributions by DRD2 gene variants to interindividual differences in vulnerability to alcoholism and polysubstance abuse will be presented. The first single gene identified for its potential contributions to polygenic disorders such as alcoholism and substance abuse, appears to deserve continuing efforts to confirm or refute its role, particularly focused at this stage on functional correlates of A1/B1-marked DRD2 gene variants.

1996 - Transcription factors: potential roles in drug-induced neuroplasticity [Articolo su rivista]
Persico, A M; Uhl, G R
abstract

Transcription factors act to regulate gene expression. Many transcription factor families have been discovered based on their roles in cell cycle events involved in development and oncogenesis. In post-mitotic neuronal cells, however, many transcription factor genes are "trans-synaptically" regulated: their patterns of expression can be dramatically altered by extracellular stimuli. Transcription factor proteins can then potently influence expression of other genes, whose products can directly alter neuronal function. The central nervous system (CNS) displays varying degrees of neuroplasticity in adult life. Flexible neurochemical pathways that link extracellular stimuli to long-term modifications in neuronal functions are likely to contribute substantially to this neuroplasticity. This review summarizes evidence supporting central roles for transcription factors in such neurochemical cascades. It furthermore illustrates how drugs of abuse can trigger and modulate neuroadaptive processes that could conceivably contribute to clinically relevant addiction phenomena such as craving, tolerance, sensitization, and withdrawal.

1995 - Brain transcription factor gene expression, neurotransmitter levels, and novelty response behaviors: Alterations during rat amphetamine withdrawal and following chronic injection stress [Articolo su rivista]
Persico, Antonio M.; Schindler, Charles W.; Zaczek, Robert; Brannock, Michael T.; Uhl, George R.
abstract

1995 - Exclusion of close linkage between the synaptic vesicular monoamine transporter locus and schizophrenia spectrum disorders [Articolo su rivista]
Persico, A. M.; Zhe Wu Wang, Null; Black, D. W.; Andreasen, N. C.; Uhl, G. R.; Crowe, R. R.
abstract

1995 - Exclusion of close linkage of the dopamine transporter gene with schizophrenia spectrum disorders [Articolo su rivista]
Persico, A; Wang Z., W; Black D., W; Andreasen N., C; Uhl G., R; Crowe, R. R.
abstract

Objective: Involvement of genetic factors in the pathogenesis of schizophrenia spectrum disorders has been indicated in twin, adoption, and familial aggregation studies; the pivotal role played by the dopamine transporter in dopaminergic neurotransmission makes it a candidate gene for these disorders. Detection of close linkage between a dopamine transporter marker and schizophrenia spectrum disorders would strongly support the existence of causal relationships between genetic mutations at the dopamine transporter locus and the disease phenotype. Method: The authors assessed the linkage between this gene and schizophrenia spectrum disorders by using polymorphic dopamine transporter gene markers in 156 subjects from 16 multiplex pedigrees with schizophrenia as well as schizophreniform, schizoaffective, and schizotypal disorders and mood-incongruent psychotic depression. Results: Complete (θ=0.0) linkage to the schizophrenia spectrum was excluded under both dominant and recessive models. Conclusions: These results indicate that allelic variants at the dopamine transporter locus do not provide major genetic contributions to the etiology of schizophrenia and related disorders in these pedigrees.

1994 - Alterations of neocortical neuronal responses to acetylcholine and GABA in rats born to alcohol-dependent mothers [Articolo su rivista]
Janiri, L; Gobbi, G; Persico, A M; Santarelli, M; Minciacchi, D; Tempesta, E
abstract

Alcohol is known to be a CNS teratogenic factor interfering with neuronal and synaptic maturation. The purpose of this microiontophoretic study was to explore GABAergic and cholinergic central mechanisms in adult rats exposed to alcohol in the third phase of prenatal life (ADM), when their mothers were subjected to alcohol physical dependence induction (9.6 g/kg/day). Responses to acetylcholine and GABA were recorded in frontal and somatosensory cortical neurons. Adult rats, whose mothers had been administered placebo with identical procedures, were used as a control (C). Cholinergic responses were significantly decreased and GABAergic responses increased in ADM animals with respect to controls. After a single i.p. alcohol injection (1.6 g/kg) spontaneous firing was depressed in ADM animals to a lesser extent than in C rats. Cholinergic excitations were reduced in C group and potentiated with reversal of atropine antagonism in ADM animals. GABAergic inhibitions were slightly increased and bicuculline antagonism was blocked in C rats, while ADM animals showed decreased responses to GABA. The present results support the hyperactivity of GABAergic system and the hypoactivity of cholinergic system reported in previous studies on prenatally and postnatally alcohol-exposed animals. Microiontophoretic results following ethanol injection led to the hypothesis that a rapid tolerance/dependence may develop in the offspring of alcohol-dependent rats.

1994 - Behavioral assessment of high-dose amphetamine withdrawal: Importance of training and testing conditions [Articolo su rivista]
Schindler, C. W.; Persico, A. M.; Uhl, G. R.; Goldberg, S. R.
abstract

Chronic d-amphetamine-treated rats were given twice daily injections at a dose of 7.5 mg/kg for 2 weeks. Acute amphetamine and saline groups of rats were given saline treatments during this time, except that for the acute group the final injection was 7.5 mg/kg d-amphetamine. Acute and chronic amphetamine groups habituated to the locomotor activity testing apparatus showed increases in both distance traveled and repetitive movement time that lasted up to 6 h following the final injection. When animals were not habituated to the activity test apparatus, however, a significant decrease in repetitive movement time was noted for the chronic amphetamine group 24-54 h following the final amphetamine injection; no differences were observed for distance traveled when the locomotor activity apparatus was novel. Swim test immobility time was assessed twice following the last injection, with the second test following the first by approximately 24 h. During the first test, decreases in immobility were observed for both chronic and acute amphetamine groups, 6-12 h following the last injection. However, during the second test, decreases in immobility time were observed only for the chronic amphetamine groups 36-72 h following the final injection. These results indicate that 24 to 72 h after the end of the chronic amphetamine regimen a withdrawal effect was observed for both repetitive movement time in the locomotor activity test and immobility time in the swim test. The withdrawal effect was observed only for the locomotor activity groups for whom the test apparatus was novel, and only during the second test of immobility time for the swim test groups. Thus, the method of behavioral assessment can be critical for the demonstration of a high-dose amphetamine withdrawal effect.

1994 - Candidate gene study of the catecholamine system in schizophrenia [Relazione in Atti di Convegno]
Crowe, R. R.; Wang, Z. W.; Black, D. W.; Andreasen, N. C.; Gelernter, J.; Persico, A
abstract

1994 - Earlier onset of psychostimulant use and the D2 dopamine receptor gene TaqI A1 RFLP predict preferential psychostimulant use in polysubstance abusers [Relazione in Atti di Convegno]
Persico, A; Bird, G.; Gabbay, F. H.; Uhl, G. R.
abstract

1994 - Genetic linkage and association studies of schizophrenia and related disorders using dopamine transporter gene markers [Relazione in Atti di Convegno]
Persico, A.; Macciardi, F.; Cavallini, M. C.; Crowe, R. R.; Wang, Z. W.; Uhl, G. R.; Smeraldi, E.
abstract

1994 - Human μ opiate receptor. cDNA and genomic clones, pharmacologic characterization and chromosomal assignment [Articolo su rivista]
Wang, Jia-Bei; Johnson, Peter S.; Persico, Antonio M.; Hawkins, Anita L.; Griffin, Constance A.; Uhl, George R.
abstract

Radioligand binding and cDNA homology studies have suggested the existence of opiate receptors distinct from the recently-cloned mu, delta and kappa receptors. XOR1S, a rat brain cDNA whose predicted translation product displays 67-72% homology with those encoded by mu 1, delta 1 and kappa 1 opiate receptor cDNAs, was constructed from two partial cDNAs identified through cDNA homology approaches. A longer XOR1L variant of this cDNA was also identified by polymerase chain reaction studies using genomic DNA and cDNA from brain and peripheral tissues. XOR1 mRNA is most highly expressed in hypothalamus. COS cell expression of both clones confers neither robust binding of opiate ligands nor reproducible opiate inhibition of forskolin-stimulated adenylate cyclase. These studies identify an orphan clone that helps to define features of the opiate receptor gene family, including apparent differential splicing and expression in peripheral tissues

1994 - Impiego della acetil-l-carnitina nella disintossicazione dalla nicotina [Brevetto]
Persico, A. M.; Malin, D. H.
abstract

1994 - Meeting report: collaborative approaches in drug abuse genetics [Articolo su rivista]
Uhl, G. R.; Persico, A.
abstract

1994 - Opiate detoxification of methadone maintenance patients using lefetamine, clonidine and buprenorphine [Articolo su rivista]
Janiri, Luigi; Mannelli, Paolo; Persico, Antonio M.; Serretti, Alessandro; Tempesta, Enrico
abstract

Thirty-nine methadone maintenance patients were included in a 9-day, double blind, randomized, inpatient detoxification trial. Methadone was tapered to 10 mg/day and then patients were assigned to one of these 3 protocols: clonidine (0.3-0.9 mg/day), lefetamine (60-240 mg/day), buprenorphine (0.15-0.9 mg/day). Buprenorphine treatment was significantly superior to clonidine and to lefetamine (F = 3.96 df = 2, 29 P &lt; 0.05) in controlling objective, subjective and psychological withdrawal symptomatology. Clonidine was more effective than lefetamine in suppressing withdrawal in the first 3 days of treatment (day 3: F = 4.10 df = 2, 30 P &lt; 0.05), and this trend was apparent on the objective and psychological items. In addition to evaluations of the efficacy of the single drugs used, the study showed that tapering methadone to low doses before entering the pharmacologically assisted discontinuation phase was clinically acceptable in detoxification from long-term methadone treatment.

1994 - cDNA Cloning of an orphan opiate receptor gene family member and its splice variant [Articolo su rivista]
Wang, Jia Bei; Johnson, Peter S.; Imai, Yasuo; Persico, Antonio M.; Ozenberger, Bradley A.; Eppler, C. Mark; Uhl, George R.
abstract

Radioligand binding and cDNA homology studies have suggested the existence of opiate receptors distinct from the recently-cloned p, S and K receptors. XORlS, a rat brain cDNA whose predicted translation product displays 67-72% homology with those encoded by ~1, 61 and ~1 opiate receptor cDNAs, was constructed from two partial cDNAs identified through cDNA homology approaches. A longer XORlL variant of this cDNA was also identified by polymerase chain reaction studies using genomic DNA and cDNA from brain and peripheral tissues. XORl mRNA is most highly expressed in hypothalamus. COS cell expression of both clones confers neither robust binding of opiate hgands nor reproducible opiate inhibition of forskolin-stimulated adenylate cyclase. These studies identify an orphan clone that helps to define features of the opiate receptor gene family, including apparent differential splicing and expression in peripheral tissues.

1994 - cDNA and genomic clones encoding human mu opiate receptor and the purified gene product [Brevetto]
Uhl, G. R.; Wang, J. B.; Johnson, P.; Persico, A. M.
abstract

A human .mu. opiate receptor cDNA has been identified from a cerebral cortical CDNA library using sequences from the rat .mu. opiate receptor CDNA. The human .mu. opiate receptor (h.mu.OR1) shares 95% amino acid identity with the rat sequence. The expressed .mu.OR1 recognizes tested opiate drugs and opioid peptides in a sodium- and GTP-sensitive fashion with affinities virtually identical to those displayed by the rat .mu. opiate receptor. Effects on cyclic AMP are similar to those noted for the rat .mu. opiate receptor. Overlapping genomic clones spanning 50 kilobasepairs and hybridizing with the h.mu.OR1 cDNA contains exon sequences encoding the entire open reading frame of the human A opiate receptor are described. Analysis of hybridization to DNA prepared from human rodent hybrid cell lines and chromosomal in situ hybridization studies indicate localization to 6q24-25. An MspI polymorphism, producing a 3.7 kb band, is being used to assess this gene’s involvement in neuropsychiatric disorders involving opiatergic systems. Claim: 1. A purified protein having the biochemical properties of a human ? opiate receptor and having an amino acid sequence comprising the amino acid sequence of SEQ ID NO:2.

1994 - cDNA and genomic clones encoding human mu opiate receptor and the purified gene product [Brevetto]
Uhl, G. R.; Wang, J. B.; Johnson, P.; Persico, A. M.
abstract

A human .mu. opiate receptor cDNA has been identified from a cerebral cortical CDNA library using sequences from the rat .mu. opiate receptor CDNA. The human .mu. opiate receptor (h.mu.OR1) shares 95% amino acid identity with the rat sequence. The expressed .mu.OR1 recognizes tested opiate drugs and opioid peptides in a sodium- and GTP-sensitive fashion with affinities virtually identical to those displayed by the rat .mu. opiate receptor. Effects on cyclic AMP are similar to those noted for the rat .mu. opiate receptor. Overlapping genomic clones spanning 50 kilobasepairs and hybridizing with the h.mu.OR1 cDNA contains exon sequences encoding the entire open reading frame of the human A opiate receptor are described. Analysis of hybridization to DNA prepared from human rodent hybrid cell lines and chromosomal in situ hybridization studies indicate localization to 6q24-25. An MspI polymorphism, producing a 3.7 kb band, is being used to assess this gene's involvement in neuropsychiatric disorders involving opiatergic systems. Claim: 1. A purified protein having the biochemical properties of a human ? opiate receptor and having an amino acid sequence comprising the amino acid sequence of SEQ ID NO:2.

1993 - A human synaptic vesicle monoamine transporter cDNA predicts posttranslational modifications, reveals chromosome 10 gene localization and identifies TaqI RFLPs [Articolo su rivista]
Surratt, C K; Persico, A M; Yang, X D; Edgar, S R; Bird, G S; Hawkins, A L; Griffin, C A; Li, X; Jabs, E W; Uhl, G R
abstract

A human vesicular monoamine transporter cDNA has been identified by screening a human brainstem library using sequences from the rat brain synaptic vesicle monoamine transporter (SVMT) [(1992) Cell 70, 539-551; (1992) Proc. Natl. Acad. Sci. USA 89, 10993-10997]. The hSVMT shares 92% amino acid identity with the rat sequence, but displays one less consensus site for asparagine N-linked glycosylation and one more consensus site for phosphorylation by protein kinase C. The human SVMT gene maps to chromosome 10q25 using Southern blotting analysis of human/rodent hybrid cell lines and fluorescent in situ hybridization approaches. The cDNA, and a subclone, recognize TaqI polymorphisms that may prove useful to assess this gene's involvement in neuropsychiatric disorders involving monoaminergic brain systems.

1993 - Brain transcription factor expression: effects of acute and chronic amphetamine and injection stress [Articolo su rivista]
Persico, A M; Schindler, C W; O'Hara, B F; Brannock, M T; Uhl, G R
abstract

Amphetamine influences behaviors and the expression of transcription factor genes in the central nervous system (CNS). A single d-amphetamine dose (7.5 mg/kg, i.p.) enhances behavioral stereotypy and augments brain expression of c-fos, fos-B, fra-1, zif268, jun-B, and c-jun by 2-11 fold. When the single amphetamine dose is preceded by 28 saline injections over 14 days, it is half as effective in enhancing expression of these genes. Rats injected with 7.5 mg/kg i.p. twice daily for 2 weeks and sacrificed after the last injection reveal further attenuation or abolition of the amphetamine-induced mRNA upregulation. These stigmata of 'tolerance' in gene expression display partial overlap with behavioral tolerance, manifest as changes in locomotor activity. Rats receiving low (2 mg/kg) amphetamine challenge doses following the 2-week 7.5 mg/kg b.i.d. amphetamine treatment show tolerance to the locomotor activating effects of the drug; no tolerance is evident following a high (7.5 mg/kg) challenge dose. These data suggest that amphetamine-induced alterations in brain transcription factor gene expression can display 'tolerance' and possibly 'cross-tolerance' with the stress caused by i.p. injection.

1993 - D2receptor gene variants and substance abuse liability [Articolo su rivista]
Persico, A. M.; Smith, S. S.; Uhl, G. R.
abstract

1993 - DOPAMINERGIC GENE-EXPRESSION DURING AMPHETAMINE WITHDRAWAL [Articolo su rivista]
Persico, A. M.; Schindler, C. W.; Brannock, M. T.; Gonzalez, A. M.; Surratt, C. K.; Uhl, G. R.
abstract

Animals and humans display a constellation of behavioral and neurochemical signs after termination of psychostimulant administration. Amphetamine withdrawal could involve the dopaminergic systems that are thought to underlie psychostimulant rewarding effects, and may thus conceivably alter expression of key genes for dopaminergic transmission, including those encoding tyrosine hydroxylase (TH), the membrane dopamine transporter (DAT) and the synaptic vesicle amine transporter (SVAT). Withdrawal from 7.5 mg kg-1 i.p. amphetamine (b.i.d. for a two week duration) yields no significant changes in rat DAT mRNA. TH mRNA levels are modestly enhanced over the same week of withdrawal, during which dopamine levels and behavioral novelty responses are both depressed. SVAT expression is significantly blunted following chronic amphetamine treatment. Altered TH and/or SVAT gene expression might contribute to restoring normal function to neurons "withdrawing" from amphetamine treatments.

1993 - Dopamine D2 receptor RFLPs, haplotypes and their association with substance use in black and caucasian research volunteers [Articolo su rivista]
O’Hara, Bruce F.; Smith, Stevens S.; Bird, Geoffrey; Persico, Antonio M.; Suarez, Brian K.; Cutting, Garry R.; Uhl, George R.
abstract

Alleles of the dopamine D2 receptor gene are distinguished by polymorphic A and B TaqI sites approximately 10 kb 3' to the final exon and bordering the second exon, respectively. These alleles have been reported to be more prevalent in heavy substance users than in control populations in several, although not all studies. Meta-analyses of combined data from available work support significant association. Two competing hypotheses could explain this association: (1) the A1 and B1 RFLPs are in linkage disequilibrium with a functional allelic determinant that in some way influences behavior; (2) the affected subjects are drawn disproportionately from populations stratified on the basis of, for example, ethnicity that happen to have higher A1 and B1 RFLP frequencies. We report here data collected from 616 substance-abusing and control individuals that document substantial differences in A1 RFLP frequencies between white and black Americans, the almost exclusive presence of the A3 RFLP in blacks, and low frequencies of rare A4 and B3 RFLPs. In blacks, neither the A1 nor B1 RFLPs display association with substance use, while white individuals display significant association with polysubstance use. When expressed as a percent of the maximum possible disequilibrium, both white and black individuals display strong linkage disequilibrium between these loci, although blacks display many more A1/B2 chromosomes. These racial differences in TaqI RFLP haplotypes underscore the need for caution in interpreting allelic associations when careful matching for ethnicity has not been performed.

1993 - Dopamine D2 receptor gene Taq I 'A' locus map including 'A4' variant: relevance for alcoholism and drug abuse [Articolo su rivista]
Persico, A. M.; O'Hara, B. F.; Farmer, S.; Gysin, R.; Flanagan, S. D.; Uhl, G. R.
abstract

The D2 dopamine receptor gene (DRD2) displays Taq I restriction fragment length polymorphisms (RFLPs) at two different loci, termed A and B. One of the three different Taq I A 'alleles' described at this site, the A1 allele (size = 6.6 kb), has been found to be associated with alcoholism and with drug abuse in the majority of studies reported to date. A complete map of the Taq I A RFLP site has been constructed, through hybridization with different fragments of the 3' flanking region and polymerase chain reaction (PCR). When screening 432 unrelated individuals to establish possible A1 allelic association with drug abuse or dependence, we have encountered a novel Taq I A RFLP, which we have named 'A4' (size = 8.6 kb). This sequence variant displays a frequency of approximately 1% in our sample and shows a Mendelizing genetic pattern in an Italian nuclear family. Primers suitable for detecting A4 using PCR have been designed. The A4, but not the A3 'allele', displays substantial overlap with the A1. In particular, A2 and A3 share the presence of a Taq I restriction site, whose absence in A1 and A4 is apparently associated with substance abuse vulnerability. Therefore, in association studies it is proper to contrast individuals displaying A1 and A4 RFLP patterns, with individuals displaying A2 and A3 RFLPs.

1993 - Dopamine transporter gene polymorphisms are not associated with polysubstance abuse [Articolo su rivista]
Persico, Antonio M.; Vandenbergh, David J.; Smith, Stevens S.; Uhl, George R.
abstract

1992 - A human dopamine transporter cDNA predicts reduced glycosylation, displays a novel repetitive element and provides racially-dimorphic TaqI RFLPs [Articolo su rivista]
Vandenbergh, D J; Persico, A M; Uhl, G R
abstract

We describe a cDNA for the human dopamine transporter, which has been implicated in several human disorders linked to dopaminergic function. The cDNA predicts reduced glycosylation of the protein with respect to the rat transporter, as well as a novel repetitive element in the 3' untranslated region of the cDNA. A TaqI RFLP is also reported that shows a race-specific difference in allelic frequencies.

1992 - Current excitement with D2 dopamine receptor gene alleles in substance abuse [Articolo su rivista]
Uhl, Gr; Persico, Am; Smith, Ss.
abstract

1992 - Genetic vulnerability to drug abuse. The D2 dopamine receptor Taq I B1 restriction fragment length polymorphism appears more frequently in polysubstance abusers [Articolo su rivista]
Smith, S S; O'Hara, B F; Persico, A M; Gorelick, D A; Newlin, D B; Vlahov, D; Solomon, L; Pickens, R; Uhl, G R
abstract

Alcoholics are more likely than nonalcoholics to display the Taq I A1 restriction fragment length polymorphism of the D2 dopamine receptor gene, according to four of six studies that examined alcoholics and controls. The current study examines whether the association observed in alcoholism might extend to other addictive substances by examining D2 dopamine receptor Taq I A and B restriction fragment length polymorphisms in polysubstance users and controls free of significant substance use. We hypothesized a stronger association for the B1 restriction fragment length polymorphism since it lies closer to dopamine receptor protein coding and 5' regulatory regions. Heavy polysubstance users and subjects with DSM-III-R psychoactive substance use diagnoses displayed significantly higher Taq I B1 frequencies than control subjects; Taq I A1 results for these comparisons were less robust. These results are consistent with a role for a D2 dopamine receptor gene variant marked by these restriction fragment length polymorphisms in enhanced substance abuse vulnerability.

1992 - Human dopamine transporter gene (DAT1) maps to chromosome 5p15.3 and displays a VNTR [Articolo su rivista]
Vandenbergh, David J.; Persico, Antonio M.; Hawkins, Anita L.; Griffin, Constance A.; Li, Xiang; Jabs, Ethylin Wang; Uhl, George R.
abstract

We describe a cDNA for the human dopamine transporter, which has been implicated in several human disorders linked to dopaminergic function. The cDNA predicts reduced glycosylation of the protein with respect to the rat transporter, as well as a novel repetitive element in the 3' untranslated region of the cDNA. A TaqI RFLP is also reported that shows a race-specific difference in allelic frequencies.

1992 - Neurotransmitter transporter family cDNAs in a rat midbrain library: 'orphan transporters' suggest sizable structural variations [Articolo su rivista]
Uhl, G R; Kitayama, S; Gregor, P; Nanthakumar, E; Persico, A; Shimada, S
abstract

1992 - Persistent decrease in heart rate after smoking cessation: A 1-year follow-up study [Articolo su rivista]
Persico, A. M.
abstract

The heart rate of 11 smokers was collected throughout their first year of abstinence. One day after smoking cessation, we recorded a significant mean heart rate drop of 9.07 beats per min, from 74.18 to 65.11 beats per min. No significant variation was afterwards detected at any chosen time point (week 1, 2, 3, 4, 6, month 3, 6, 12). In fact, 1 year after cessation the mean heart rate was still 66.36 beats per min, well below initial baseline values. These data indicate that the decrease in heart rate following smoking cessation mostly represents a permanent return to individual normal values in the absence of nicotine self-administration. Nonetheless, three subjects did show a trend toward the recovery of their precessation heart rate and one subject fully recovered it between months 3 and 6 of abstinence. This trend suggests that, while heart rate adaptation to nicotine is largely of an acute nature, there may also be a chronic component. Its role, though usually minor, should become detectable in a few subjects, because of wide interindividual variability.

1992 - Predictors of smoking cessation in a sample of Italian smokers [Articolo su rivista]
Persico, A M
abstract

In this study we identify several pretreatment characteristics which predict abstinence at 6 months. Moreover, the persistence of withdrawal discomfort and of an increased frequency of night awakenings during the first month of abstinence, together with a tendency to "slip" during Weeks II-IV, strongly predicted relapse. Our results suggest that: 1) Predictors of outcome cannot be automatically extended from one cultural context to another; 2) a careful assessment of certain variables, made while the patient is still under treatment, provides significant prognostic hints; 3) ex-smokers' sleeping and dreaming function has been ignored by the literature, whereas they may well be involved into the maintenance of the drug-free state.

1992 - Sequence of human dopamine transporter cDNA [Brevetto]
Uhl, G. R.; Vandenbergh, D. J.; Persico, A. M.
abstract

The cloning and characterization of a human dopamine transporter (HUDAT) cDNA is described. RFLP analysis is used to determine the distribution of HUDAT alleles in two ethnic backgrounds. The means by which the association between HUDAT alleles and behavioral disorders which have altered HUDAT expression as a basis for their etiology is discussed. Methods for evaluating the expression of HUDAT are described.

1992 - Sequence of human dopamine transporter cDNA [Brevetto]
Uhl, G. R.; Vandenbergh, D. J.; Persico, A. M.
abstract

The cloning and characterization of a human dopamine transporter (HUDAT) cDNA is described. RFLP analysis is used to determine the distribution of HUDAT alleles in two ethnic backgrounds. The means by which the association between HUDAT alleles and behavioral disorders which have altered HUDAT expression as a basis for their etiology is discussed. Methods for evaluating the expression of HUDAT are described.

1991 - A prospective assessment of opiate addiction treatment protocols for inpatients with HIV-related syndromes [Articolo su rivista]
Persico, Antonio M.; Di Giannantonio, Massimo; Tempesta, Enrico
abstract

A sample of 114 intravenous drug abusers hospitalized for HIV-spectrum diseases, was allowed to choose between methadone maintenance, partial or complete withdrawal as inpatient programs and was reassessed 4 days after discharge. One third of the patients had relapsed into heroin abuse. Rates of early relapse were significantly lower in the partial withdrawal group and higher in the methadone maintenance group. Patients with longer hospitalization and more severe HIV-related syndromes were particularly at risk of early relapse. Also, 45 former intravenous drug abusers were reassessed. Relapses were even more frequent than among active abusers, especially if the drug-free period before admission had been shorter than 1 year.

1991 - Activity of L-carnitine and L-acetylcarnitine on cholinoceptive neocortical neurons of the rat in vivo [Articolo su rivista]
Janiri, L; Falcone, M; Persico, A; Tempesta, E
abstract

Carnitine and acetylcarnitine have been demonstrated to be present in the CNS and to be involved in cholinergic mechanisms, even if their exact role in neurotransmission is still unknown. This microiontophoretic study was carried out on single cholinoceptive neurons of the somatosensory cortex in the rat in order to analyze the effects of L- and D-carnitine and L-acetylcarnitine on the spontaneous firing and the neuronal responses to some putative transmitters. L-carnitine and L-acetylcarnitine increased the spontaneous discharge rate, while D-carnitine was found to be ineffective. L-acetylcarnitine clearly potentiated the cholinergic excitatory responses. On the contrary, L-carnitine was found to reduce cholinergic responses in a great percentage of units and to inhibit L-acetylcarnitine-induced excitatory responses. Atropine blocked the increase in firing rate induced by L-carnitine and L-acetylcarnitine, thus suggesting for both of them a muscarinic activity. No interactions were observed between carnitines and GABA and glutamate. These results show that carnitine and acetylcarnitine are stereospecific neuroactive compounds with a cholinomimetic activity. They may play a role in a modulatory system for the cholinoceptive cortical neuron.

1991 - Aids and psychiatric disorders: Guidelines for psychopharmacological treatment [Articolo su rivista]
Persico, A.; Di Giannantonio, M.; Mattioni, T.; Lestingi, L.; Zeppetelli, E.; Tempesta, E.
abstract

1991 - Benzodiazepines consumption and risk of dependence in institutionalized geriatric patients [Articolo su rivista]
Janiri, L.; Di Giovanni, A.; Persico, A; Zeppetelli, E.; Mannelli, P.; Antico, L.; Tempesta, E.
abstract

The use of psychoactive drugs in geriatric nursing homes has to be considered within the peculiar frame work of the elderly condition. The present study was aimed at verifying the prevalence and characteristics of prolonged benzodiazepine use (greater than 1 month) in the entire elderly institutionalized population of an Italian region (Molise). In the 31 benzodiazepine consumers identified, psychosocial and clinical data were recorded by means of a specific scale and of a semistructured interview for the assessment of drug effects. The dependence potential was evaluated in a smaller sample of 18 patients, recording with a scale the onset of withdrawal and/or rebound symptoms following discontinuation of drug treatment. The characteristics of prescription appeared to be generally correct and no appreciable tendency toward abuse was evidenced. In only one patient, discontinuation of benzodiazepines precipitated a confusional state. In other 5 patients significant but not severe withdrawal symptoms were recorded. Duration of treatment was the only variable moderately correlated with the onset of withdrawal/rebound symptomatology (p less than 0.1). In conclusion, the results of our study are reassuring but they should not induce to disregard important clinical aspects of psychopharmacotherapy in the elderly, such as: psychopathological diagnosis, drug associations and interactions, duration of treatment and dosage of prescribed benzodiazepines.

1991 - Zipeprol is a newly abused antitussive with an opioid spectrum and hallucinogenic effects [Articolo su rivista]
Janiri, Luigi; Mannelli, Paolo; Persico, Antonio M.; Diodato, Silvana; Tempesta, Enrico
abstract

Zipeprol is a non-opioid cough suppressor agent recently abused in Italy. Clinical reports from 30 street abusers show that the drug in high doses has a definite abuse liability and dependence potential. Many of its effects were identical to those induced by opiates, although specific dysperceptive symptoms were commonly reported by patients. Spontaneous withdrawal symptoms were similar to those of opiates. Withdrawal could be also precipitated by naloxone. It is concluded that zipeprol can induce dependence with a mechanism which may be mediated by some types of opioid receptors.

1990 - Clinical reports on recent abuse of an antitussive [Articolo su rivista]
Tempesta, E; Janiri, L; Mannelli, P; Persico, A M; Diodato, S
abstract

1989 - Dual effects of lephetamine on spontaneous and evoked neuronal firing in the somatosensory cortex of the rat [Articolo su rivista]
Janiri, L.; Persico, A. M.; Tempesta, E.
abstract

Lephetamine is a central analgesic, recently shown to be abused by drug addicts and to induce dependence in humans. The drug was applied microiontophoretically on single neurones of the somatosensory cortex of the rat in vivo. Its activity on the spontaneous and evoked firing rate was recorded. Morphine and naloxone were employed to verify the hypothesis that a mu-opiate mechanism of action could be involved. The most frequent response evoked by lephetamine was a dose-dependent excitation non-reversible by naloxone. On the other hand, units inhibited or apparently unaffected by the drug, showed a selective anti-glutamate (and partly anti-acetylcholine) effect, which was reversed by either systemically- or iontophoretically-administered naloxone. Long-lasting (8-12 min) applications of lephetamine caused a progressive desensitization of cortical neurones to the inhibitory and anti-glutamate effect. The inhibitory activity of lephetamine and morphine was additive and an increased neuronal excitability was shown by a post-inhibitory rebound of glutamate-induced neuronal activity. The action exerted by lephetamine on glutamate-induced excitations and on postsynaptic excitability, its reversibility by naloxone and the occurrence of acute tolerance allow the conclusion that only the inhibitory effect of lephetamine is mediated by an opioid mechanism. The lephetamine-induced excitations, not reversed by naloxone, are difficult to interpret as opioid-mediated.

1989 - The biological basis of psychoimmunology [Articolo su rivista]
Persico, A.; Janiri, L.; Tempesta, E.
abstract

1988 - Genetic studies of panic disorder and related conditions [Capitolo/Saggio]
Crowe, R. R.; Noyes, R.; Persico, A. M.; Wilson, A. F.; Elston, R. C.
abstract

1987 - Pro-opiomelanocortin (POMC) gene excluded as a cause of panic disorder in a large family [Articolo su rivista]
Crowe, R R; Noyes, R; Persico, A M
abstract