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Silvia ALBONI

Professore Associato
Dipartimento di Scienze della Vita sede ex-Scienze Biomediche

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Pubblicazioni

2023 - ADOLESCENCE AS A CRITICAL TIME-WINDOW FOR NEUROINFLAMMATION IN THE MOUSE: WHY SEX MATTERS [Abstract in Rivista]
Toscano, Ylenia; Benatti, Cristina; Alboni, Silvia; Ciani, Miriam; Rigillo, Giovanna; Tascedda, Fabio; Blom, Johanna Maria Catharina; Brunello, Nicoletta
abstract

2023 - Environmental enrichment influences mouse hippocampal neuroinflammatory response [Abstract in Rivista]
Rigillo, Giovanna; Benatti, Cristina; Toscano, Ylenia; Tascedda, Fabio; Pani, Luca; Brunello, Nicoletta; Alboni, Silvia
abstract

Background: Many studies have shown the positive effects of environmental enrichment on brain plasticity with significant implications for development, behavior, learning, memory, and recovery from brain damage (1). Experimental and clinical studies report that one’s living environment can modulate cellular and molecular responses in the brain, counteracting cognitive decline, alleviating anxiety, and depressive behaviours, as well as moderating the outcome of pharmacological treatments (2). Neuroinflammation has been well established as an important factor in the aetiopathogenesis and progression of brain disorders. It can affect neural development and alters hippocampal plasticity thus resulting in cognitive impairments. Neuroinflammation is characterized by a dysregulation of the NLRP3 inflammasome activation, an increase in the expression of inflammatory cytokines, and a decrease of neurotrophic factors (3). Behavioral and neurochemical changes, caused by neuroinflammation, have been most frequently investigated through peripheral administration of lipopolysaccharide (LPS), which can, directly and indirectly, affect the central nervous system (4). Based on these premises, the aim of this study was to explore the molecular effects of the quality of the living environment in modulating the LPS-induced neuroinflammatory response in the hippocampus of wild-type mice. Methods: Male C57BL6J mice (13 weeks-old) were randomly housed in Impoverished (IE) or Enriched Environment (EE) condition for 28 days, then exposed to LPS (0.830 mg/Kg, i.p.) or saline (SAL). Twenty-four hours after injection hippocampi were removed for gene expression analysis performed by means of qRT-PCR. Data from groups were analyzed by Two-way ANOVA followed by Tukey’s post hoc test. Results: The analysis of the environmental effects on the LPS signaling system highlighted the downregulation of the membrane-bound protein LBP, the receptor TLR4 and the co-receptor cluster CD14 expression levels in EE-housed animals compared to their counterparts. The exposure to an EE condition was able to attenuate the LPS-induced increase of TLR4 and NLRP3 inflammasome mRNA levels. Gene expression analysis revealed a significant downregulation of the pro-inflammatory cytokines IL-1β and TNFα levels in EE-housed mice while LPS exposure strongly increased IL-1β and TNFα mRNA levels irrespective of the housing conditions. Moreover, EE-exposed mice showed a significant upregulation of BDNF hippocampal mRNA levels, although no effects were observed after LPS treatment in both conditions. Conclusions: Our results displayed the beneficial effect of EE in regulating the expression of inflammatory mediators involved in the LPS-induced response in the hippocampus, a key area for learning, memory, and emotion. These data suggest that living environment may exert a positive and protective role on the brain by reducing susceptibility toward neurodegenerative or neuropsychiatric disorders.

2023 - Farmaci antidepressivi parte II: SNRI e farmaci multimodali [Capitolo/Saggio]
Benatti, Cristina; Alboni, Silvia; Brunello, Nicoletta; Pani, Luca; Tascedda, Fabio
abstract

2023 - Hydroxypropyl-β-Cyclodextrin Depletes Membrane Cholesterol and Inhibits SARS-CoV-2 Entry into HEK293T-ACEhi Cells [Articolo su rivista]
Alboni, Silvia; Secco, Valentina; Papotti, Bianca; Vilella, Antonietta; Pia Adorni, Maria; Zimetti, Francesca; Schaeffer, Laurent; Tascedda, Fabio; Zoli, Michele; Leblanc, Pascal; Villa, Erica
abstract

2023 - VORTIOXETINE ATTENUATES NEUROINFLAMMATION BY MODULATING THE NOD-LIKE RECEPTOR FAMILY PYRIN DOMAIN CONTAINING 3 INFLAMMASOME ACTIVATION IN MICROGLIA: IMPLICATIONS FOR COGNITIVE FUNCTION [Abstract in Rivista]
Rigillo, G.; Ciani, M.; Benatti, C.; Blom, J. M. C.; Tascedda, F.; Pani, L.; Alboni, S.; Brunello, N.
abstract

Vortioxetine (VTX) is a multimodal antidepressant with an extensive pharmacological profile that includes modulation of various neurotransmitter systems, neuroprotective activity, and beneficial effects on cognitive functions. Recent research has revealed a novel aspect of VTX's activity - its antiinflammatory effects - that suggests an intriguing molecular mechanism may underpin its therapeutic benefits. Neuroinflammation, dysfunctional neurogenesis and neurotransmission, and dysregulation of the hypothalamus– pituitary–adrenal (HPA) axis are all pivotal in the onset and progression of depression. One particular immune-inflammatory pathway overactivated in brain disorders is the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, a multiprotein complex. This complex's activation is mediated by NF-kB and reactive oxygen species (ROS) signalling pathways, leading to caspase-1-dependent release of the proinflammatory cytokines, IL-1β and IL-18. Mounting evidence implicates NLRP3 inflammasome in neuroinflammation- related disorders, with its activation associated with cognitive function impairment. Of note, microglia, the resident immune cells crucial for brain plasticity, express high levels of the NLRP3-inflammasome components. Our initial findings indicate that VTX exerts a region-dependent modulatoryeffect on the NLRP3-inflammasome system in a LPS-induced memory impairment in vivo model. Furthermore, VTX's ability to modulate immune response suggests that microglia could be a direct target of the drug. AIM: In light of the compelling evidence surrounding the role of the NLRP3 inflammasome in cognitive dysfunctions and the recent discovery of VTX's anti-inflammatory activity, we aimed to investigate the molecular effects induced by VTX pre-treatment in the presence or absence of the inflammasome-inducer LPS in a well-established in vitro model of mouse microglia: BV2 cells. METHODS: To dissect the influence of VTX pre-treatment (24h) on the NLRP3 inflammasome signaling pathway and microglial polarization, we analyzed gene and protein expression in BV2 cells stimulated with LPS or vehicle for 6h. We also scrutinized the activation/translocation of NF-kB and ROS release under these conditions. We applied one-way or two-way ANOVA followed by Tukey’s post hoc test for statistical analysis based on the experimental design. RESULTS: Our data demonstrate that short-term exposure to LPS significantly induces the activation/translocation of NF-kB signaling and ROS release in BV2 cells. We observed a time-dependent transcriptional upregulation of the inflammasome complex, IL-1β and IL-18, and microglial pro-inflammatory targets post-LPS stimulation, alongside a downregulation of the anti-inflammatory factors. Interestingly, a pre-treatment with VTX (10 nM) for 24h effectively modulated the LPS-induced NF-kB translocation and ROS production compared to control cells. Cells pre-treated with VTX exhibited lower levels of LPS-induced NLRP3 inflammasome- and microglia pro-inflammatory-related targets. However, VTX did not influence the expression of anti-inflammatory factors in both unstimulated and LPS-stimulated BV2 cells. CONCLUSIONS: Our findings reinforce the emerging evidence that supports VTX's anti-inflammatory activity. This activity is mediated via modulation of the inflammasome signaling pathway, which plays a pivotal role in the inflammatory response of microglia cells.

2022 - Deciphering the central immunomodulatory effects of a vortioxetine pretreatment on the LPS-induced inflammatory cascade [Abstract in Atti di Convegno]
Ciani, M.; Toscano, Y.; Benatti, C.; Blom, J. M. C.; Tascedda, F.; Alboni, S.; Brunello, N.
abstract

2022 - Depressive disorders [Capitolo/Saggio]
Ferrari, S; Blanch, J; Lavasani, S; Beall, Sc; Gibson, Sj; Magarini, Fm; Alboni, S
abstract

Major Depressive Disorder (MDD) and other disorders on the depressive spectrum are more prevalent among HIV-infected individuals than in the general population. These illnesses have a significant impact on prevention, engagement and adherence to HIV care, prognosis, illness-related disability, and quality of life. Diagnosis of depressive disorders may be delayed, and management may be inadequate, due to interfering factors such as overlapping symptoms, co-morbid neurocognitive disorders, and risk of drug-drug interactions. Not only do HIV infection and disorders on the depressive spectrum act as mutually reciprocal risk factors, but their concomitance may provide an etiopathogenetic model; co-morbid depressive disorders may contribute to understanding the complex role played by the neuro-endocrine-immune system in the development, maintenance, and chronicity of psychiatric symptoms. Many neurobiological pathways are involved, including the HPA axis and the gut microbiome. Various therapeutic options are available for MDD among HIV-infected patients, including antidepressant medications, other psychopharmacological interventions, and non-pharmacological strategies. Antidepressants are safe and appropriate for the care of persons with HIV and co-morbid depressive disorders. The recognition and treatment of depressive symptoms and disorders among HIV-infected persons is a major clinical goal, considering the high prevalence of this co-morbidity and its impact on prognosis and perceived quality of life.

2022 - Destabilizers of the thymidylate synthase homodimer accelerate its proteasomal degradation and inhibit cancer growth [Articolo su rivista]
Costantino, Luca; Ferrari, Stefania; Santucci, Matteo; MH Salo-Ahen, Outi; Carosati, Emanuele; Franchini, Silvia; Lauriola, Angela; Pozzi, Cecilia; Trande, Matteo; Gozzi, Gaia; Saxena, Puneet; Cannazza, Giuseppe; Losi, Lorena; Cardinale, Daniela; Venturelli, Alberto; Quotadamo, Antonio; Linciano, Pasquale; Tagliazucchi, Lorenzo; Moschella, MARIA GAETANA; Guerrini, Remo; Pacifico, Salvatore; Luciani, Rosaria; Genovese, Filippo; Henrich, Stefan; Alboni, Silvia; Santarem, Nuno; CORDEIRO DA SILVA, Anabela; Giovannetti, Elisa; J Peters, Godefridus; Pinton, Paolo; Rimessi, Alessandro; Cruciani, Gabriele; M Stroud, Robert; C Wade, Rebecca; Mangani, Stefano; Marverti, Gaetano; D'Arca, Domenico; Ponterini, Glauco; Costi, Maria Paola
abstract

2022 - DIFFERENZE DI GENERE NELLA COMORBILITÀ TRA SINTOMATOLOGIA ANSIOSO-DEPRESSIVA, SINDROME METABOLICA E ADENOMI COLORETTALI [Abstract in Atti di Convegno]
Rioli, Giulia; Bonamici, Caterina; Mancini, Stefano; Mattei, Giorgio; Alboni, Silvia; Sena, Paola; Roncucci, Luca; Fiore, Gianluca; Pingani, Luca; Ferrari, Silvia; Galeazzi, Gian Maria
abstract

2022 - Differenze di genere nella comorbilità tra sintomatologia ansioso-depressiva, sindrome metabolica e adenomi colorettali [Abstract in Rivista]
Rioli, G.; Bonamici, C.; Mancini, S.; Mattei, G.; Alboni, S.; Sena, P.; Roncucci, L.; Fiore, G.; Pingani, L.; Ferrari, S.; Galeazzi, G. M.
abstract

2022 - Gender differences in Anxious-depressive symptomatology, Metabolic Syndrome and Colorectal Adenomas among outpatients undergoing colonoscopy: a cross-sectional study according to a PNEI perspective [Articolo su rivista]
Rioli, Giulia; Mattei, Giorgio; Bonamici, Caterina; Mancini, Stefano; Alboni, Silvia; Cannazza, Giuseppe; Sena, Paola; Roncucci, Luca; Pingani, Luca; Ferrari, Silvia; Galeazzi, Gian Maria
abstract

2022 - Gut microbiota alterations promote traumatic stress susceptibility associated with p-cresol-induced dopaminergic dysfunctions [Articolo su rivista]
Laudani, Samuele; Torrisi, Sebastiano A; Alboni, Silvia; Bastiaanssen, Thomaz F S; Benatti, Cristina; Rivi, Veronica; Moloney, Rachel D; Fuochi, Virginia; Furneri, Pio M; Drago, Filippo; Salomone, Salvatore; Tascedda, Fabio; Cryan, John F; Leggio, Gian Marco
abstract

Mounting evidence suggests a link between gut microbiota abnormalities and post-traumatic stress disorder (PTSD). However, whether and how the gut microbiota influences PTSD susceptibility is poorly understood. Here using the arousal-based individual screening model, we provide evidence for pre-trauma and post-trauma gut microbiota alterations in susceptible mice exhibiting persistent PTSD-related phenotypes. A more in-depth analysis revealed an increased abundance of bacteria affecting brain processes including myelination, and brain systems like the dopaminergic neurotransmission. Because dopaminergic dysfunctions play a key role in the pathophysiological mechanisms subserving PTSD, we assessed whether these alterations in gut microbiota composition could be associated with abnormal levels of metabolites inducing dopaminergic dysfunctions. We found high levels of the l-tyrosine-derived metabolite p-cresol exclusively in the prefrontal cortex of susceptible mice. We further uncovered abnormal levels of dopamine and DOPAC, together with a detrimental increase of dopamine D3 receptor expression, exclusively in the prefrontal cortex of susceptible mice. Conversely, we observed either resilience mechanisms aimed at counteracting these p-cresol-induced dopaminergic dysfunctions or myelination-related resilience mechanisms only in the prefrontal cortex of resilient mice. These findings reveal that gut microbiota abnormalities foster trauma susceptibility and thus it may represent a promising target for therapeutic interventions.

2022 - Identification and characterization of the kynurenine pathway in the pond snail Lymnaea stagnalis [Articolo su rivista]
Benatti, Cristina; Rivi, Veronica; Alboni, Silvia; Grilli, Andrea; Castellano, Sara; Pani, Luca; Brunello, Nicoletta; Blom, Johanna Maria Catharina; Bicciato, Silvio; Tascedda, Fabio
abstract

2022 - Il ruolo dell'infiammazione sistemica cronica e della via metabolica delle chinurenine nella comorbidilità tra ansia, depressione e sindrome metabolica: risultati di uno studio cross-sectional [Abstract in Rivista]
Rioli, G.; Bonamici, C.; Macini, S.; Mattei, G.; Alboni, S.; Sena, P.; Roncucci, L.; Fiore, G.; Pingani, L.; Ferrari, S.; Galeazzi, G. M.
abstract

2022 - IL RUOLO DELL’INFIAMMAZIONE SISTEMICA CRONICA E DELLA VIA METABOLICA DELLE CHINURENINE NELLA COMORBIDITÀ TRA ANSIA, DEPRESSIONE E SINDROME METABOLICA: RISULTATI DI UNO STUDIO CROSS-SECTIONAL [Abstract in Atti di Convegno]
Rioli, Giulia; Bonamici, Caterina; Macini, Stefano; Mattei, Giorgio; Alboni, Silvia; Sena, Paola; Roncucci, Luca; Fiore, Gianluca; Pingani, Luca; Ferrari, Silvia; Galeazzi, Gian Maria
abstract

2022 - Indoleamine 2, 3-Dioxygenase 1 Mediates Survival Signals in Chronic Lymphocytic Leukemia via Kynurenine/Aryl Hydrocarbon Receptor-Mediated MCL1 Modulation [Articolo su rivista]
Atene, C. G.; Fiorcari, S.; Mesini, N.; Alboni, S.; Martinelli, S.; Maccaferri, M.; Leonardi, G.; Potenza, L.; Luppi, M.; Maffei, R.; Marasca, R.
abstract

The indoleamine 2,3-dioxygenase 1 (IDO1) metabolic circuitry, comprising the first tryptophan (Trp) catabolite L-kynurenine (Kyn) and the aryl hydrocarbon receptor (AHR), has emerged as a mechanism of cancer immune evasion. Here, we investigated the functional role of the IDO1/Kyn/AHR axis in chronic lymphocytic leukemia (CLL). Our data show that CLL cells expressed an active form of the IDO1 enzyme and microenvironmental stimuli can positively modulate its expression. Interferon (IFN)-γ induces IDO1 expression through the Jak/STAT1 pathway and mediates Kyn production concomitantly with Trp consumption in CLL-conditioned media, while INCB018424 (ruxolitinib), a JAK1/2 inhibitor, impaired both effects. To characterize the involvement of IDO1 in leukemic cell maintenance, we overexpressed IDO1 by vector transfection measuring enhanced resistance to spontaneous apoptosis. IDO1 pro-survival influence was confirmed by treating CLL cells with Kyn, which mediated the increase of induced myeloid leukemia cell differentiation protein (MCL1). Conversely, AHR silencing or its blockade via CH-223191 improved the apoptosis of leukemic clones and mitigated MCL1 expression. Moreover, Kyn-treated CLL cells are less affected by the pro-apoptotic effect of ABT-199 (venetoclax), while CH-223191 showed synergistic/additive cytotoxicity with this drug. Lastly, targeting directly MCL1 in CLL cells with AMG-176, we abrogate the pro-survival effect of Kyn. In conclusion, our data identify IDO1/Kyn/AHR signaling as a new therapeutic target for CLL, describing for the first time its role in CLL pathobiology.

2022 - Microglial diversity along the hippocampal longitudinal axis impacts synaptic plasticity in adult male mice under homeostatic conditions [Articolo su rivista]
De Felice, E; Gonçalves de Andrade, E; Golia, M T; González Ibáñez, F; Khakpour, M; Di Castro, M A; Garofalo, S; Di Pietro, E; Benatti, C; Brunello, N; Tascedda, F; Kaminska, B; Limatola, C; Ragozzino, D; Tremblay, M E; Alboni, S; Maggi, L
abstract

The hippocampus is a plastic brain area that shows functional segregation along its longitudinal axis, reflected by a higher level of long-term potentiation (LTP) in the CA1 region of the dorsal hippocampus (DH) compared to the ventral hippocampus (VH), but the mechanisms underlying this difference remain elusive. Numerous studies have highlighted the importance of microglia-neuronal communication in modulating synaptic transmission and hippocampal plasticity, although its role in physiological contexts is still largely unknown. We characterized in depth the features of microglia in the two hippocampal poles and investigated their contribution to CA1 plasticity under physiological conditions. We unveiled the influence of microglia in differentially modulating the amplitude of LTP in the DH and VH, showing that minocycline or PLX5622 treatment reduced LTP amplitude in the DH, while increasing it in the VH. This was recapitulated in Cx3cr1 knockout mice, indicating that microglia have a key role in setting the conditions for plasticity processes in a region-specific manner, and that the CX3CL1-CX3CR1 pathway is a key element in determining the basal level of CA1 LTP in the two regions. The observed LTP differences at the two poles were associated with transcriptional changes in the expression of genes encoding for Il-1, Tnf-α, Il-6, and Bdnf, essential players of neuronal plasticity. Furthermore, microglia in the CA1 SR region showed an increase in soma and a more extensive arborization, an increased prevalence of immature lysosomes accompanied by an elevation in mRNA expression of phagocytic markers Mertk and Cd68 and a surge in the expression of microglial outward K+ currents in the VH compared to DH, suggesting a distinct basal phenotypic state of microglia across the two hippocampal poles. Overall, we characterized the molecular, morphological, ultrastructural, and functional profile of microglia at the two poles, suggesting that modifications in hippocampal subregions related to different microglial statuses can contribute to dissect the phenotypical aspects of many diseases in which microglia are known to be involved.

2022 - Non-psychotropic Cannabis sativa L. phytocomplex modulates microglial inflammatory response through CB2 receptors-, endocannabinoids-, and NF-κB-mediated signaling [Articolo su rivista]
Borgonetti, V.; Benatti, C.; Governa, P.; Isoldi, G.; Pellati, F.; Alboni, S.; Tascedda, F.; Montopoli, M.; Galeotti, N.; Manetti, F.; Miraldi, E.; Biagi, M.; Rigillo, G.
abstract

Cannabis sativa L. is increasingly emerging for its protective role in modulating neuroinflammation, a complex process orchestrated among others by microglia, the resident immune cells of the central nervous system. Phytocannabinoids, especially cannabidiol (CBD), terpenes, and other constituents trigger several upstream and downstream microglial intracellular pathways. Here, we investigated the molecular mechanisms of a CBD- and terpenes-enriched C. sativa extract (CSE) in an in vitro model of neuroinflammation. We evaluated the effect of CSE on the inflammatory response induced by exposure to lipopolysaccharide (LPS) in BV-2 microglial cells, compared with CBD and β-caryophyllene (CAR), CB2 receptors (CB2r) inverse and full agonist, respectively. The LPS-induced upregulation of the pro-inflammatory cytokines IL-1β, IL-6, and TNF-α was significantly attenuated by CSE and only partially by CBD, whereas CAR was ineffective. In BV-2 cells, these anti-inflammatory effects exerted by CSE phytocomplex were only partially dependent on CB2r modulation and they were mediated by the regulation of enzymes responsible for the endocannabinoids metabolism, by the inhibition of reactive oxygen species release and the modulation of JNK/p38 cascade with consequent NF-κB p65 nuclear translocation suppression. Our data suggest that C. sativa phytocomplex and its multitarget mechanism could represent a novel therapeutic strategy for neuroinflammatory-related diseases.

2022 - Stress, Hormones, and Metabolism [Capitolo/Saggio]
Radighieri, Giulia; Alboni, Silvia
abstract

From an evolutionary perspective, the stress response system has evolved to ensure the survival of the organism and therefore of the species. Triggering the stress response system leads to several hormone-mediated metabolic alterations, aimed at exploiting energy resources and adaptive behavioral strategies to cope with real or perceived threats. In the modern highly stressed society, the majority of the stressors we face chronically are psychological. The consequent activation of the stress response system mobilizes energy resources that are not expended in the short term, but rather compromise the energetic balance of the organism for long intervals. Stress and obesity are widespread conditions that mutually interplay, as they share at the basis an intricated network of pathways that regulate the physical, behavioral, and cognitive spheres of the organism. The type of stressor, along with substantial differences in genetic, environmental, and developmental factors, are important determinants for the vulnerability to hormones-related negative consequences on metabolism, and therefore the different susceptibility to develop obesity and eventually metabolic diseases associated with an obese state, like metabolic syndrome and type 2 diabetes. The main purpose of this article is to discuss the cardinal stress-activated pathways that, combined with an impaired energy balance, result in fat mass accumulation, especially visceral, and in the onset of a chronic low-grade inflammation. These features contribute to the development of insulin resistance, and therefore to the incidence of common metabolic dysregulations which in turn represent important risk factors for other widespread and severe pathologies, such as cardiovascular diseases and tumors.

2021 - Preliminary results of a multidisciplinary italian study adopting a psycho-neuro-endocrine-immunological (Pnei) approach to the study of colorectal adenomas [Articolo su rivista]
Mancini, S.; Alboni, S.; Mattei, G.; Rioli, G.; Sena, P.; Marchi, M.; Sacchetti, A.; Boarino, V.; Roncucci, L.; Galeazzi, G. M.; Ferrari, S.
abstract

Background and aim of the work: Colorectal mucosal precancerous lesions, metabolic syndrome (MetS) and psychiatric disorders may share a common low-grade local and systemic inflammation. Aim is to report on preliminary data concerning a research adopting a psycho-neuro-endocrine-immune (PNEI) approach to study outpatients undergoing colonoscopy. Methods: A sample of patients undergoing colonosco-py was cross-sectionally investigated. Data on colorectal adenomas, MetS, early atherosclerosis, anxious-de-pressive symptoms, personality traits, and inflammatory markers were statistically analyzed. Results: Sixty-two patients were recruited (female 50%, mean age: 60.8±9.4 years). The prevalence of adenomas and MetS was respectively of 45.2% and 41.9%. Anxiety and depressive symptoms were detected in 16 (32.7%) and 9 (18.4%) subjects, respectively. The presence of adenomas positively correlated with male sex (p=0.01), age (p<0.01), IL-6 (p=0.03), hsCRP (p=0.04), and MetS (p=0.03); it was also associated with hsCRP concentra-tion (aOR=3.81, p=0.03). Conclusions: Proinflammatory atherogenic status, psychological traits, increased mucosal inflammation, and metabolic parameters may share a common a pathogenic mechanism, worth studying.

2021 - Serum metabolic signature of binge-like palatable food consumption in female rats by nuclear magnetic resonance spectroscopy [Articolo su rivista]
Cifani, Carlo; Alboni, Silvia; Mucci, Adele; Benatti, Cristina; Botticelli, Luca; Brunello, Nicoletta; Micioni Di Bonaventura, Maria Vittoria; Righi, Valeria
abstract

Maladaptive eating behavior is a growing public health problem and compulsively eating excessive food in a short time, or binge eating, is a key symptom of many eating disorders. In order to investigate the binge-like eating behavior in female rats, induced by intermittent food restrictions/refeeding and frustration stress, we analyzed for the first time the metabolic profile obtained from serum of rats, through nuclear magnetic resonance (NMR) spectroscopy. In this experimental protocol, rats were exposed to chow food restricting/refeeding and frustration stress manipulation. This stress procedure consists of 15 min exposure to the odor and sight of a familiar chocolate paste, without access to it, just before offering the palatable food. In this model, a "binge-eating episode" was considered the significantly higher palatable food consumption within 2 h in restricted and stressed rats (R + S) than in the other three experimental groups: rats with no food restriction and no stress (NR + NS), only stressed rats (NR + S) or only restricted rats (R + NS). Serum samples from these four different rat groups were collected. The statistical analysis of the 1 H NMR spectral profiles of the four sets of samples pointed to O- and N-acetyl glycoproteins as the main biomarkers for the discrimination of restriction effects. Other metabolites, such as threonine, glycine, glutamine, acetate, pyruvate and lactate, showed trends that may be useful to understand metabolic pathways involved in eating disorders. This study suggested that NMR-based metabolomics is a suitable approach to detect biomarkers related to binge-eating behavior.

2021 - Vortioxetine Prevents Lipopolysaccharide-Induced Memory Impairment Without Inhibiting the Initial Inflammatory Cascade [Articolo su rivista]
Alboni, S.; Benatti, C.; Colliva, C.; Radighieri, G.; Blom, J. M. C.; Brunello, N.; Tascedda, F.
abstract

Vortioxetine is a novel multimodal antidepressant that modulates a wide range of neurotransmitters throughout the brain. Preclinical and clinical studies have shown that vortioxetine exerts positive effects on different cognitive domains and neuroprotective effects. Considering the key role of microglial cells in brain plasticity and cognition, we aimed at investigating the effects of pretreatment with vortioxetine in modulating behavioral and molecular effects induced by an immune challenge: peripheral injection of lipopolysaccharide (LPS). To this purpose, C57BL/6J male mice were first exposed to a 28-day standard diet or vortioxetine-enriched diet, which was followed by an acute immune challenge with LPS. Sickness symptoms and depressive-like behaviors (anhedonia and memory impairment) were tested 6 and 24 h after exposure to LPS, respectively. Moreover, the expressions of markers of immune activation and M1/M2 markers of microglia polarization were measured in the dorsal and ventral parts of the hippocampus. The pretreatment with vortioxetine did not affect both LPS-induced sickness behavior and anhedonia but prevented the deficit in the recognition memory induced by the immune challenge. At the transcriptional level, chronic exposure to vortioxetine did not prevent LPS-induced upregulation of proinflammatory cytokines 6 h after the immune challenge but rather seemed to potentiate the immune response to the challenge also by affecting the levels of expression of markers of microglia M1 phenotype, like cluster of differentiation (CD)14 and CD86, in an area-dependent manner. However, at the same time point, LPS injection significantly increased the expression of the M2 polarization inducer, interleukin 4, only in the hippocampus of animals chronically exposed to vortioxetine. These results demonstrate that a chronic administration of vortioxetine specifically prevents LPS-induced memory impairment, without affecting acute sickness behavior and anhedonia, and suggest that hippocampal microglia may represent a cellular target of this novel antidepressant medication. Moreover, we provide a useful model to further explore the molecular mechanisms specifically underlying cognitive impairments following an immune challenge.

2021 - What can we teach Lymnaea and what can Lymnaea teach us? [Articolo su rivista]
Rivi, Veronica; Benatti, Cristina; Lukowiak, Ken; Colliva, Chiara; Alboni, Silvia; Tascedda, Fabio; Blom, Johanna M. C.
abstract

2019 - Combined fluoxetine and metformin treatment potentiates antidepressant efficacy increasing IGF2 expression in the dorsal hippocampus [Articolo su rivista]
Poggini, S.; Golia, M. T.; Alboni, S.; Milior, G.; Sciarria, L. P.; Viglione, A.; Bon, G. M.; Brunello, N.; Puglisi-Allegra, S.; Limatola, C.; Maggi, L.; Branchi, I.
abstract

An increasing number of studies show that selective serotonin reuptake inhibitors (SSRIs) exert their therapeutic action, at least in part, by amplifying the influence of the living environment on mood. As a consequence, when administered in a favorable environment, SSRIs lead to a reduction of symptoms, but in stressful conditions, they show limited efficacy. Therefore, novel therapeutic approaches able to neutralize the influence of the stressful environment on treatment are needed. The aim of our study was to test whether, in a mouse model of depression, the combined administration of SSRI fluoxetine and metformin, a drug able to improve the metabolic profile, counteracts the limited efficacy of fluoxetine alone when administered in stressful conditions. Indeed, metabolic alterations are associated to both the onset of major depression and the antidepressant efficacy. To this goal, adult C57BL/6 male mice were exposed to stress for 6 weeks; the first two weeks was aimed at generating a mouse model of depression. During the remaining 4 weeks, mice received one of the following treatments: vehicle, fluoxetine, metformin, or a combination of fluoxetine and metformin. We measured liking- and wanting-type anhedonia as behavioral phenotypes of depression and assessed the expression levels of selected genes involved in major depressive disorder and antidepressant response in the dorsal and ventral hippocampus, which are differently involved in the depressive symptomatology. The combined treatment was more effective than fluoxetine alone in ameliorating the depressive phenotype after one week of treatment. This was associated to an increase in IGF2 mRNA expression and enhanced long-term potentiation, specifically in the dorsal hippocampus, at the end of treatment. Overall, the present results show that, when administered in stressful conditions, the combined fluoxetine and metformin treatment may represent a more effective approach than fluoxetine alone in a short term. Finally, our findings highlight the relevance of polypharmacological strategy as effective interventions to increase the efficacy of the antidepressant drugs currently available.

2019 - Interplay between inflammation and neural plasticity: Both immune activation and suppression impair LTP and BDNF expression [Articolo su rivista]
Golia, M. T.; Poggini, S.; Alboni, S.; Garofalo, S.; Ciano Albanese, N.; Viglione, A.; Ajmone-Cat, M. A.; St-Pierre, A.; Brunello, N.; Limatola, C.; Branchi, I.; Maggi, L.
abstract

An increasing number of studies show that both inflammation and neural plasticity act as key players in the vulnerability and recovery from psychiatric disorders and neurodegenerative diseases. However, the interplay between these two players has been limitedly explored. In fact, while a few studies reported an immune activation, others conveyed an immune suppression, associated with an impairment in neural plasticity. Therefore, we hypothesized that deviations in inflammatory levels in both directions may impair neural plasticity. We tested this hypothesis experimentally, by acute treatment of C57BL/6 adult male mice with different doses of two inflammatory modulators: lipopolysaccharide (LPS), an endotoxin, and ibuprofen (IBU), a nonselective cyclooxygenase inhibitor, which are respectively a pro- and an anti-inflammatory agent. The results showed that LPS and IBU have different effects on behavior and inflammatory response. LPS treatment induced a reduction of body temperature, a decrease of body weight and a reduced food and liquid intake. In addition, it led to increased levels of inflammatory markers expression, both in the total hippocampus and in isolated microglia cells, including Interleukin (IL)-1β, and enhanced the concentration of prostaglandin E2 (PGE2). On the other hand, IBU increased the level of anti-inflammatory markers, decreased tryptophan 2,3-dioxygenase (TDO2), the first step in the kynurenine pathway known to be activated during inflammatory conditions, and PGE2 levels. Though LPS and IBU administration differently affected mediators related with pro- or anti-inflammatory responses, they produced overlapping effects on neural plasticity. Indeed, higher doses of both LPS and IBU induced a statistically significant decrease in the amplitude of long-term potentiation (LTP), in Brain-Derived Neurotrophic Factor (BDNF) expression levels and in the phosphorylation of the AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptor subunit GluR1, compared to the control group. Such effect appears to be dose-dependent since only the higher, but not the lower, dose of both compounds led to a plasticity impairment. Overall, the present findings indicate that acute treatment with pro- and anti-inflammatory agents impair neural plasticity in a dose dependent manner.

2019 - Modulation of neuroplasticity-related targets following stress-induced acute escape deficit [Articolo su rivista]
Benatti, C.; Radighieri, Giulia; Alboni, S.; Blom, J. M. C.; Brunello, N.; Tascedda, F.
abstract

Understanding resilience is a major challenge to improve current pharmacological therapies aimed at complementing psychological-based approaches of stress-related disorders. In particular, resilience is a multi-factorial construct where the complex network of molecular events that drive the process still needs to be resolved. Here, we exploit the acute escape deficit model, an animal model based on exposure to acute unavoidable stress followed by an escape test, to define vulnerable and resilient phenotypes in rats. Hippocampus and prefrontal cortex (PFC), two of the brain areas most involved in the stress response, were analysed for gene expression at two different time points (3 and 24 h) after the escape test. Total Brain-Derived Neurotrophic Factor (BDNF) was highly responsive in the PFC at 24-h after the escape test, while expression of BDNF transcript IV increased in the hippocampus of resistant animals 3 h post-test. Expression of memory enhancers like Neuronal PAS Domain Protein 4 (Npas4) and Activity-regulated cytoskeleton-associated protein (Arc) decreased in a time- and region-dependent fashion in both behavioural phenotypes. Also, the memory inhibitor Protein Phosphatase 1 (Ppp1ca) was increased in the hippocampus of resilient rats at 3 h post-test. Given the importance of neurotrophic factors and synaptic plasticity-related genes for the development of appropriate coping strategies, our data contribute to an additional step forward in the comprehension of the psychobiology of stress and resiliency.

2019 - The Association Between Symptoms of Anxiety, Depression, and Cardiovascular Risk Factors: Results From an Italian Cross-Sectional Study [Articolo su rivista]
Rioli, Giulia; Tassi, Silvia; Mattei, Giorgio; Ferrari, Silvia; Galeazzi, Gian Maria; Mancini, Stefano; Alboni, Silvia; Roncucci, Luca
abstract

Cardiovascular diseases, anxiety, and depression are among the most frequent clinical conditions in the Western world, often in comorbidity. Evidence regarding a shared pathophysiology suggests a mediating role by chronic systemic inflammation. The aims of this study were to measure the association between anxiety and depressive symptoms, cardiovascular risk factors, and inflammatory markers. Outpatients aged 40 years or more undergoing colonoscopy after positive fecal occult blood test were enrolled; the following data were collected: body mass index, blood pressure, blood glucose, lipid profile, C-reactive protein (CRP) level, carotid thickness, Hospital Anxiety and Depression Scale, Temperament and Character Inventory, INTERdisciplinary MEDicine Self-Assessment, and 36-Item Short-Form Health Survey scores. Fifty-four patients were enrolled; 30.2% had anxiety symptoms, 18.9% depressive symptoms, and 9.4% concomitant anxiety-depressive symptoms. Anxiety symptoms were associated with low high-density lipoprotein levels. Depressive symptoms were associated with CRP levels, providing supporting evidence for the role of inflammation in the pathophysiology of depression.

2018 - Molecular changes associated with escitalopram response in a stress-based model of depression [Articolo su rivista]
Benatti, Cristina; Alboni, Silvia; Blom, Johanna Maria Catharina; Mendlewicz, Julien; Tascedda, Fabio; Brunello, Nicoletta
abstract

Converging evidence points at hypothalamus-pituitary-adrenal (HPA) axis hyperactivity and neuroinflammation as important factors involved in the etiopathogenesis of major depressive disorder (MDD) and in therapeutic efficacy of antidepressants. In this study, we examined the molecular effects associated with a response to a week-long treatment with escitalopram in the chronic escape deficit (CED) model, a validated model of depression based on the induction of an escape deficit after exposure of rats to an unavoidable stress. We confirmed our previous result that a treatment with escitalopram (10 mg/kg) was effective after 7 days in reverting the stress-induced escape deficit in approximately 50% of the animals, separating responders from non-responders. Expression of markers of HPA axis functionality as well as several inflammatory mediators were evaluated in the hypothalamus, a key structure integrating signals from the neuro, immune, endocrine systems. In the hypothalamus of responder animals we observed a decrease in the expression of CRH and its receptors and an increase in GR protein in total and nuclear extracts; this effect was accompanied by a significant decrease in circulating corticosterone in the same cohort. Hypothalamic IL-1β and TNFα expression were increased in stressed animals, while CXCL2, IL-6, and ADAM17 mRNA levels were decreased in escitalopram treated rats regardless of the treatment response. These data suggest that efficacy of a one week treatment with escitalopram may be partially mediated by a decrease HPA axis activity, while in the hypothalamus the drug-induced effects on the expression of immune modulators did not correlate with the behavioural outcome.

2018 - Neither all anti-inflammatory drugs nor all doses are effective in accelerating the antidepressant-like effect of fluoxetine in an animal model of depression [Articolo su rivista]
Alboni, Silvia; Benatti, Cristina; Capone, Giacomo; Tascedda, Fabio; Brunello, Nicoletta
abstract

Introduction: Non-steroidal anti-inflammatory drugs (NSAIDs) have been studied as possible adjunctive therapy in the treatment of depression. However, administering NSAIDs to increase the effectiveness of antidepressant has yielded inconsistent results. Methods: We evaluated the effect of the co-administration of fluoxetine (5 mg/kg) and flurbiprofen (5 mg/kg) or fluoxetine (5 mg/kg) and celecoxib (5 mg/kg) in the chronic escape deficit (CED) model of depression after 7 days of treatment. The co-administration of fluoxetine plus acetylsalicylic acid (ASA, 45 mg/kg i.p.) was used as a positive control. Moreover, we tested the behavioral effect of different doses (45, 22.5, and 11.25 mg/Kg i.p.) of ASA as potentiating agent of the effect of fluoxetine in the same paradigm. Results: Our study showed that only the co-administration of ASA with fluoxetine was able to revert the stress-induced condition of escape deficit after 7 days of treatment, and that the amplitude of the antidepressant-like effect of ASA was dose dependent. In the same experimental conditions, celecoxib with fluoxetine only partially resolved the stress-induced impaired behavior while flurbiprofen/fluoxetine cotreatment was ineffective. Limitations: Our study is still exploratory, more doses, longer treatment regimens, and different behavioral outcomes must be investigated to draw a clear conclusion. Conclusion: Our results further stress the importance of the type and dose when NSAIDs are associated with antidepressants to ameliorate a clinical response.

2017 - An Italian observational study on subclinical cardiovascular risk factors and depressive symptomatology. A suggestion for the potential utility of a sinergic cardio-psychiatric perspective [Abstract in Rivista]
Tassi, S.; Rioli, Giulia; Mattei, Giorgio; Mancini, Stefano; Alboni, Silvia; Roncucci, Luca; Sena, Paola; Mariani, Francesco; Marchi, Mattia; Fabbrizi, Andrea; Feltri, L.; Visentini, Chiara; Pollutri, Gabriella; Artoni, Cecilia; Saraceni, Serena; Galli, Giacomo; Spiga, Giulia; Minarini, Alessandro; Perrone, Daniela; Galletti, Martina; Giambalvo, Nina; Montardi, Giulia; Galeazzi, Gian Maria; Ferrari, Silvia
abstract

Introduction Growing evidence has been collected over the complex, intertwined pathophysiological connection among subclinical cardiovascular (CV) disease, i.e. atherosclerosis, systemic low pro-inflammatory states and psychiatric disorders/symptomatology (anxiety, depression), with controversial results. Aim Aim of this study was to investigate the possible link between subclinical CV risk factors (atherosclerosis), depressive symptoms, and inflammation. Methods Cross-sectional study. Inclusion criteria: outpatients aged ≥40 years, attending colonoscopy after positive faecal occult blood test, negative medical history for cancer. Collected data: blood pressure, glycaemia, lipid profile, waist circumference, BMI, PCR (C reactive protein), LPS (bacterial lipopolysaccharide), ultrasound carotid intima-media thickness (c-IMT). Psychometric tests: HADS, TCI, IMSA, SF36. Statistical analysis performed with STATA13. Results The 54 patients enrolled were equally distributed by gender. CV risk factors were common in the study population, with 33 patients (61.11%) with hypertension, 14 (25.93%) with hyperglycaemia, 20 (37.4%) with hypertriglyceridemia, 19 (35.19%) with low HDL and 64.81% with overweight. High levels of PCR were found in 24 subjects (44.44%). Right c-IMT was increased in 26.41% of the sample, and 11.32% had an atheromatous plaque. Left c-IMT was increased in 24.53% of patients, with a plaque in 7.55% of them. Clinically relevant depressive symptoms were found in the 18.87% of the sample and were statistically significantly associated with PCR (OR = 28.63; P = 0.01). Conclusions Evidence contributing to the so-called “inflammation theory” of depression and supporting the association between mood and CV disorders was here collected, supporting the need for a multidisciplinary approach to the diagnosis and treatment of such conditions, assuming a clinically-translated PNEI (psycho-neuro-endocrino-immunological) perspective.

2017 - Cardiovascular risk factors, anxiety symptoms and inflammation markers: Evidence of association from a cross-sectional study [Abstract in Rivista]
Rioli, Giulia; Tassi, S.; Mattei, Giorgio; Alboni, Silvia; Mancini, Stefano; Artoni, Cecilia; Galletti, Martina; Giambalvo, Nina; Galli, Giacomo; Marchi, Mattia; Minarini, Alessandro; Montardi, Giulia; Perrone, Daniela; Pollutri, Gabriella; Roncucci, Luca; Saraceni, Serena; Spiga, Giulia; Visentini, Chiara; Galeazzi, Gian Maria; Ferrari, Silvia
abstract

Introduction Anxiety disorders and Cardiovascular (CV) diseases, among the most common disorders in Western World, are often comorbid. A chronic systemic inflammatory state might be a shared underlining pathophysiological mechanism. Aims To investigate the association between anxiety symptoms, CV risks factors and inflammatory markers in an outpatient sample. Methods Cross-sectional study. Inclusion criteria: outpatients aged ≥40 years, attending colonoscopy after positive faecal occult blood test, negative medical history for cancer. Collected data: blood pressure, glycaemia, lipid profile, waist circumference, BMI, PCR (C Reactive Protein), LPS (bacterial Lipopolysaccharide). Psychometric tests: HADS, TCI, IMSA, SF36. Statistical analysis performed with STATA13. Results Fifty four patients enrolled (27 males, 27 females). Sixteen patients (30.19%) were positive for anxiety symptoms. Thirty-three patients (61.11%) had hypertension, 14 (25.93%) hyperglycaemia and 64.81% were overweight, with frank obesity (BMI≥ 30) in 11 subjects (20.37%). Anxiety symptoms were associated with low hematic HDL values (OR = 0.01; P = 0.01) and high concentration of triglycerides (OR = 0.023; P = 0.02) at the multiple regression model. At the univariate logistic analysis, anxiety was associated with LPS (OR = 1.06; P = 0.04). Conclusions Further evidence over the epidemiological link between common mental disorders and CV diseases was collected, with possible hints on pathophysiology and causative mechanisms related to inflammation. The importance of screening for anxiety and depression in medical populations is confirmed. Suggestions on future availability of screening tools based on inflammatory-related indicators should be the focus of future research.

2017 - Fluoxetine effects on molecular, cellular and behavioral endophenotypes of depression are driven by the living environment [Articolo su rivista]
Alboni, Silvia; van Dijk, R. M.; Poggini, Silvia; Milior, G.; Perrotta, M.; Drenth, T.; Brunello, Nicoletta; Wolfer, D. P.; Limatola, C.; Amrein, I.; Cirulli, F.; Maggi, L.; Branchi, I.
abstract

Selective serotonin reuptake inhibitors (SSRIs) represent the most common treatment for major depression. However, their efficacy is variable and incomplete. In order to elucidate the cause of such incomplete efficacy, we explored the hypothesis positing that SSRIs may not affect mood per se but, by enhancing neural plasticity, render the individual more susceptible to the influence of the environment. Consequently, SSRI administration in a favorable environment promotes a reduction of symptoms, whereas in a stressful environment leads to a worse prognosis. To test such hypothesis, we exposed C57BL/6 mice to chronic stress in order to induce a depression-like phenotype and, subsequently, to fluoxetine treatment (21 days), while being exposed to either an enriched or a stressful condition. We measured the most commonly investigated molecular, cellular and behavioral endophenotypes of depression and SSRI outcome, including depression-like behavior, neurogenesis, brain-derived neurotrophic factor levels, hypothalamic-pituitary–adrenal axis activity and long-term potentiation. Results showed that, in line with our hypothesis, the endophenotypes investigated were affected by the treatment according to the quality of the living environment. In particular, mice treated with fluoxetine in an enriched condition overall improved their depression-like phenotype compared with controls, whereas those treated in a stressful condition showed a distinct worsening. Our findings suggest that the effects of SSRI on the depression- like phenotype is not determined by the drug per se but is induced by the drug and driven by the environment. These findings may be helpful to explain variable effects of SSRI found in clinical practice and to device strategies aimed at enhancing their efficacy by means of controlling environmental conditions.

2017 - Hippocampus-related effects of fluoxetine treatment under stressful vs enriched conditions [Articolo su rivista]
Alboni, S.; van Dijk, M.; Poggini, S.; Milior, G.; Perrotta, M. L.; Drenth, T.; Brunello, N.; Wolfer, D.; Limatola, C.; Amrein, I.; Cirulli, F.; Maggi, L.; Branchi, I.
abstract

2017 - How are personality traits and physical activity involved in colorectal carcinogenesis? A cross-sectional study on patients undergoing colonoscopy [Abstract in Rivista]
Marchi, Mattia; Alboni, Silvia; Artoni, Cecilia; Galletti, Martina; Giambalvo, Nina; Mancini, Stefano; Mariani, Francesco; Mattei, Giorgio; Merighi, A.; Montardi, Giulia; Pollutri, Gabriella; Rioli, Giulia; Saraceni, Serena; Spiga, Giulia; Visentini, Chiara; Ferrari, Silvia
abstract

Introduction Inflammatory state of the large bowel is a key factor for the development of colorectal cancer (CRC). It has multifactorial aetiology, including psychological determinants. Physical activity may have a protective function against CRC via anti-inflammatory properties; on the contrary, personality traits correlate with an unhealthy and dangerous lifestyle. Objective To measure the association between personality traits, lifestyle and colonoscopy outcome. Methods Cross sectional study. Patients undergoing colonoscopy aged 40 or more, with a negative history for cancer or inflammatory bowel disease, were enrolled. Data collected: colonoscopy outcome, smoke, alcohol, physical activity, presence/absence of Metabolic Syndrome, personality traits assessed by the Temperament & Character Inventory (TCI). Results In a sample of 53 subjects (females = 24, 45.3%), the mean age was 60.66 ± 9.08. At least one adenoma was found to 23 patients (43.3%). Twenty patients were smokers (37.74%), 36 (67.92%) drank alcohol at least weekly; approximately 60% reported regular physical activity. At the multivariate regression, the outcome was associated to: TCI Self Transcendence domain (ST) (OR = 1.36, P = 0.04) and physical activity (OR = 0.14, P = 0.03). Conclusion People with ST's characteristic personality traits and sedentary life style are more likely to have precancerous colorectal lesions. This confirms the protective role of physical activity, and suggests to further explore the role of personality in cancerogenesis.

2017 - Hypothalamic expression of inflammatory mediators in an animal model of binge eating [Articolo su rivista]
Alboni, Silvia; Micioni di Bonaventura, Maria Vittoria; Benatti, Cristina; Giusepponi, Maria; Elena, ; Brunello, Nicoletta; Cifani, Carlo
abstract

Binge eating episodes are characterized by uncontrollable, distressing eating of a large amount of highly palatable food and represent a central feature of bingeing related eating disorders. Research suggests that inflammation plays a role in the onset and maintenance of eating-related maladaptive behavior. Markers of inflammation can be selectively altered in discrete brain region where they can directly or indirectly regulate food intake. In the present study we measured expression levels of different components of cytokine systems (IL-1, IL-6, IL-18, TNF- and IFN-ɣ) and related molecules (iNOS and COX2) in the preoptic and anterior-tuberal parts of the hypothalamus of a validated animal model of binge eating. In this animal model, based on the exposure to both food restriction and frustration stress, binge-like eating behavior for highly palatable food is not shown when animals are exposed to the frustration stress during the estrus phase. We found a characteristic down-regulation of the IL-18/ IL-18 receptor system (with increased expression of the inhibitor of the pro-inflammatory cytokine IL18, IL-18BP, together with a decreased expression of the binding chain of the IL-18 receptor) and a three-fold increase in the expression of iNOS specifically in the anterior- tuberal region of the hypothalamus of animals that develop a binge-like eating behavior. Differently, when food restricted animals were stressed during the estrus phase IL-18 expression increased while iNOS expression was not significantly affected. Considering the role of this region of the hypothalamus in controlling feeding related behavior, this can be relevant in eating disorders and obesity. Our data suggest that by targeting centrally selected inflammatory markers, we may prevent that disordered eating turns into a full blown eating disorder.

2017 - Prevalence of metabolic syndrome and of symptoms of anxiety and depression in patients undergoing colonoscopy [Abstract in Rivista]
Marchi, Mattia; Alboni, Silvia; Fabbrizi, Andrea; Feltri, L.; Galli, Giacomo; Guicciardi, Alessia; Mancini, Stefano; Mattei, Giorgio; Minarini, Alessandro; Perrone, Daniela; Rioli, Giulia; Roncucci, Luca; Sena, Paola; Ferrari, Silvia
abstract

Introduction Metabolic syndrome (MetS) is defined by metabolic and cardio-vascular impairments and is frequently associated with anxiety and depressive disorders. Both MetS and anxiety-depressive syndromes feature similar systemic inflammatory alterations. Inflammation of the large bowel is also a key factor for the development of colorectal cancer (CRC). Objective To measure the prevalence of MetS and symptoms of anxiety and depression among patients undergoing colonoscopy. Methods Cross-sectional study. Patients undergoing colonoscopy aged 40 or more, with negative history for neoplasia or inflammatory bowel disease, were enrolled. Data collected: colonoscopy outcome, presence/absence of MetS (IDF and ATP III criteria), presence/absence of depressive and anxiety symptoms assessed with HADS. Results The sample was made up of 53 patients (female 24, 45.3%). Mean age was 60.66 ± 9.08. At least one adenoma was found to 23 patients (43.3%). Prevalence of MetS ranged from 34% to 36% (ATP III and IDF criteria, respectively). Prevalence of depressive and anxiety symptoms was 20% and 33%, respectively. Conclusion Prevalence of MetS, anxiety and depressive symptoms among patients undergoing colonoscopy was higher than in the general population.

2017 - Rescue of IL-1β-induced reduction of human neurogenesis by omega-3 fatty acids and antidepressants [Articolo su rivista]
Borsini, Alessandra; Alboni, Silvia; Horowitz, Mark A.; Tojo, Luis M.; Cannazza, Giuseppe; Su, Kuan-Pin; Pariante, Carmine M.; Zunszain, Patricia A.
abstract

Both increased inflammation and reduced neurogenesis have been associated with the pathophysiology of major depression. We have previously described how interleukin-1 (IL-1) β, a pro-inflammatory cytokine increased in depressed patients, decreases neurogenesis in human hippocampal progenitor cells. Here, using the same human in vitro model, we show how omega-3 (ω-3) polyunsaturated fatty acids and conventional antidepressants reverse this reduction in neurogenesis, while differentially affecting the kynurenine pathway. We allowed neural cells to proliferate for 3 days and further differentiate for 7 days in the presence of IL-1β (10 ng/ml) and either the selective serotonin reuptake inhibitor sertraline (1 µM), the serotonin and norepinephrine reuptake inhibitor venlafaxine (1 µM), or the ω-3 fatty acids eicosapentaenoic acid (EPA, 10 µM) or docosahexaenoic acid (DHA, 10 µM). Co-incubation with each of these compounds reversed the IL-1β-induced reduction in neurogenesis (DCX- and MAP2-positive neurons), indicative of a protective effect. Moreover, EPA and DHA also reversed the IL-1β-induced increase in kynurenine, as well as mRNA levels of indolamine-2,3-dioxygenase (IDO); while DHA and sertraline reverted the IL-1β-induced increase in quinolinic acid and mRNA levels of kynurenine 3-monooxygenase (KMO). Our results show common effects of monoaminergic antidepressants and ω-3 fatty acids on the reduction of neurogenesis caused by IL-1β, but acting through both common and different kynurenine pathway-related mechanisms. Further characterization of their individual properties will be of benefit towards improving a future personalized medicine approach.

2017 - The screening for depression and neurocognitive disorders in subjects newly diagnosed with HIV [Abstract in Rivista]
Ferrari, Silvia; Piemonte, Chiara; Feltri, L.; Ottolini, Fedra; Maffei, Simone; Nanni, M. G.; Alboni, Silvia
abstract

Background Inflammatory mediators may be relevant to explain the frequent comorbidity between depression, neurocognitive disorders and HIV. HIV induces activation of inflammatory mediators, mainly cytokines, that have been involved in the onset of depression and response to antidepressant treatment. Aim To identify recurring profiles of inflammatory biomarkers subtending depression, effectiveness of antidepressants and neurocognitive disorders among HIV-infected individuals. Methods All adult newly HIV-diagnosed out-patients attending HIV clinics in three towns of Northern Italy were screened, assessed for depression and studied immunologically and for neurocognitive disorders. Results Twenty-five patients have been enrolled so far: of these, 35% were positive to PHQ-9 screening, of which 6 were positive to the diagnostic assessment for depression. No neurocognitive disorders were found among the patients. As the project will develop, it is expected that frequency of depression, neurocognitive disorders and effective antidepressant treatment will be found to correlate to the profile of immune biomarkers. These findings might help to understand the etiology of depression in HIV, and specifically the role of inflammation and immunological changes.

2016 - Changes in the NMR Metabolic Profile of Live Human Neuron-Like SH-SY5Y Cells Exposed to Interferon-α2 [Articolo su rivista]
Valeria, Righi; Schenetti, Luisa; Mucci, Adele; Benatti, Stefania; Tascedda, Fabio; Brunello, Nicoletta; Carmine, Pariante M; Alboni, Silvia
abstract

Interferon (IFN)-α2 is an extensively therapeutically used pro-inflammatory cytokine. Though its efficacy in controlling viral replication and tumor cells proliferation, administration of IFN-α2 is often associated with the development of central side effects. Magnetic resonance spectroscopy studies have demonstrated that IFN-α2 administration affects brain metabolism, however the exact nature of this effect is not completely known. We hypothesized that IFN-α2 can affect metabolic activity of human neuron-like SH-SY5Y cells which possess many characteristics of neurons and represent one of the most used models for studying mechanisms involved in neurotoxicity or neuroprotection. To test our hypothesis we have characterized the metabolic signature of live SH-SY5Y, and their conditioned media, after 24 and 72 h of exposure to vehicle or IFN-α2 (100 ng/ml) by using High Resolution-Magic Angle Spinning (HR-MAS) Nuclear Magnetic Resonance (NMR) spectroscopy. Our results revealed that 1) the use of HR-MAS NMR is ideally suitable for the characterization of the metabolic profile of live cells and their conditioned media without extraction procedures; and 2) a 72 h exposure to IFN-α2 increases the level of metabolites involved in maintaining energetic (including creatine and lactate) and osmotic (such as myo-inositol, scyllo-inositol, taurine and glycerophosphorylcholine) balances in neuron-like cells and of metabolic waste products (namely lactate, ethanol and acetate), glycine and glutamine in their growth media. These results may contribute to gain more knowledge about the IFN-α2 induced effect on the brain and support the interpretation of magnetic resonance spectroscopy studies performed in humans.

2016 - Disease-induced neuroinflammation and depression [Articolo su rivista]
Benatti, Cristina; Blom, Johanna Maria Catharina; Rigillo, Giovanna; Alboni, Silvia; Zizzi, Francesca; Torta, Riccardo; Brunello, Nicoletta; Tascedda, Fabio
abstract

Progression of major depression, a multifactorial disorder with a neuroinflammatory signature, seems to be associated with the disruption of body allostasis. High rates of comorbidity between depression and specific medical disorders, such as, stroke, chronic pain conditions, diabetes mellitus, and human immunodeficiency virus (HIV) infection, have been extensively reported. In this review, we discuss how these medical disorders may predispose an individual to develop depression by examining the impact of these disorders on some hallmarks of neuroinflammation known to be impaired in depressed patients: altered permeability of the blood brain barrier, immune cells infiltration, activated microglia, increased cytokines production, and the role of inflammasomes. In all four pathologies, blood brain barrier integrity was altered, allowing the infiltration of peripheral factors, known to activate resident microglia. Evidence indicated morphological changes in the glial population, increased levels of circulating pro-inflammatory cytokines or increased production of these mediators within the brain, all fundamental in neuroinflammation, for the four medical disorders considered. Moreover, activity of the kynurenine pathway appeared to be enhanced. With respect to the inflammasome NLRP3, a new target whose role in neuroinflammation is emerging as being important, accumulating data suggest its involvement in the pathogenesis of brain injury following stroke, chronic pain conditions, diabetes mellitus or in HIV associated immune impairment. Finally, data gathered over the last 10 years, indicate and confirm that depression, stroke, chronic pain, diabetes, and HIV infection share a combination of underlying molecular, cellular and network mechanisms leading to a general increase in the neuroinflammatory burden for the individual.

2016 - Editorial: Cytokines as players of neuronal plasticity and sensitivity to environment in healthy and pathological brain [Articolo su rivista]
Alboni, Silvia; Maggi, Laura
abstract

NA

2016 - Erratum to: Changes in the NMR Metabolic Profile of Live Human Neuron-Like SH-SY5Y Cells Exposed to Interferon-α2 (J Neuroimmune Pharmacol, 2015, DOI 10.1007/s11481-015-9641-x) [Altro]
Righi, Valeria; Schenetti, Luisa; Mucci, Adele; Benatti, Stefania; Tascedda, Fabio; Brunello, Nicoletta; Pariante, Carmine M.; Alboni, Silvia
abstract

Erratum

2016 - Fluoxetine treatment affects the inflammatory response and microglial function according to the quality of the living environment [Articolo su rivista]
Alboni, Silvia; Poggini, Silvia; Garofalo, Stefano; Milior, Giampaolo; El Hajj, Hassan; Lecours, Cynthia; Girard, Isabelle; Gagnon, Steven; Boisjoly Villeneuve, Samuel; Brunello, Nicoletta; Wolfer, David P; Limatola, Cristina; Tremblay, Marie Ève; Maggi, Laura; Branchi, Igor
abstract

It has been hypothesized that selective serotonin reuptake inhibitors (SSRIs), the most common treatment for major depression, affect mood through changes in immune function. However, the effects of SSRIs on inflammatory response are contradictory since these act either as anti- or pro-inflammatory drugs. Previous experimental and clinical studies showed that the quality of the living environment moderates the outcome of antidepressant treatment. Therefore, we hypothesized that the interplay between SSRIs and the environment may, at least partially, explain the apparent incongruence regarding the effects of SSRI treatment on the inflammatory response. In order to investigate such interplay, we exposed C57BL/6 mice to chronic stress to induce a depression-like phenotype and, subsequently, to fluoxetine treatment or vehicle (21days) while being exposed to either an enriched or a stressful condition. At the end of treatment, we measured the expression levels of several anti- and pro-inflammatory cytokines and inflammatory mediators in the whole hippocampus and in isolated microglia. We also determined microglial density, distribution, and morphology to investigate their surveillance state. Results show that the effects of fluoxetine treatment on inflammation and microglial function, as compared to vehicle, were dependent on the quality of the living environment. In particular, fluoxetine administered in the enriched condition increased the expression of pro-inflammatory markers compared to vehicle, while treatment in a stressful condition produced anti-inflammatory effects. These findings provide new insights regarding the effects of SSRIs on inflammation, which may be crucial to devise pharmacological strategies aimed at enhancing antidepressant efficacy by means of controlling environmental conditions.

2016 - Fractalkine receptor deficiency impairs microglial and neuronal responsiveness to chronic stress [Articolo su rivista]
Milior, Giampaolo; Lecours, Cynthia; Samson, Louis; Bisht, Kanchan; Poggini, Silvia; Pagani, Francesca; Deflorio, Cristina; Lauro, Clotilde; Alboni, Silvia; Limatola, Cristina; Branchi, Igor; Tremblay, Marie Eve; Maggi, Laura
abstract

Chronic stress is one of the most relevant triggering factors for major depression. Microglial cells are highly sensitive to stress and, more generally, to environmental challenges. However, the role of these brain immune cells in mediating the effects of stress is still unclear. Fractalkine signaling - which comprises the chemokine CX3CL1, mainly expressed by neurons, and its receptor CX3CR1, almost exclusively present on microglia in the healthy brain - has been reported to critically regulate microglial activity. Here, we investigated whether interfering with microglial function by deleting the Cx3cr1 gene affects the brain's response to chronic stress. To this purpose, we housed Cx3cr1 knockout and wild-type adult mice in either control or stressful environments for 2weeks, and investigated the consequences on microglial phenotype and interactions with synapses, synaptic transmission, behavioral response and corticosterone levels. Our results show that hampering neuron-microglia communication via the CX3CR1-CX3CL1 pathway prevents the effects of chronic unpredictable stress on microglial function, short- and long-term neuronal plasticity and depressive-like behavior. Overall, the present findings suggest that microglia-regulated mechanisms may underlie the differential susceptibility to stress and consequently the vulnerability to diseases triggered by the experience of stressful events, such as major depression.

2016 - The proinflammatory cytokine interleukin 18 regulates feeding by acting on the bed nucleus of the stria terminalis [Articolo su rivista]
Francesconi, Walter; Sánchez Alavez, Manuel; Berton, Fulvia; Alboni, Silvia; Benatti, Cristina; Mori, Simone; Nguyen, William; Zorrilla, Eric; Moroncini, Gianluca; Tascedda, Fabio; Conti, Bruno
abstract

The proinflammatory cytokine IL-18 has central anorexigenic effects and was proposed to contribute to loss of appetite observed during sickness. Here we tested in the mouse the hypothesis that IL-18 can decrease food intake by acting on neurons of the bed nucleus of the stria terminalis (BST), a component of extended amygdala recently shown to influence feeding via its projections to the lateral hypothalamus (LH). We found that both subunits of the heterodimeric IL-18 receptor are highly expressed in the BST and that local injection of recombinant IL-18 (50 ng/ml) significantly reduced c-fos activation and food intake for at least 6 h. Electrophysiological experiments performed in BST brain slices demonstrated that IL-18 strongly reduces the excitatory input on BST neurons through a presynaptic mechanism. The effects of IL-18 are cell-specific and were observed in Type III but not in Type I/II neurons. Interestingly, IL-18-sensitve Type III neurons were recorded in the juxtacapsular BST, a region that contains BST-LH projecting neurons. Reducing the excitatory input on Type III GABAergic neurons, IL-18 can increase the firing of glutamatergic LH neurons through a disinhibitory mechanism. Imbalance between excitatory and inhibitory activity in the LH can induce changes in food intake. Effects of IL-18 were mediated by the IL-18R because they were absent in neurons from animals null for IL-18Rα (Il18ra-/-), which lack functional IL-18 receptors. In conclusion, our data show that IL-18 may inhibit feeding by inhibiting the activity of BST Type III GABAergic neurons.

2015 - P.2.a.012 Co-administration of fluoxetine with acetylsalicylic acid, but not flurbiprofen or celecoxib, for one week shows an antidepressant-like effect [Abstract in Rivista]
Alboni, Silvia; Benatti, Cristina; Tascedda, Fabio; Brunello, Nicoletta
abstract

Increasing evidence is now demonstrating the involvement of the immune system and in particular of their effectors, cytokines, in the development and progression of depression. In particular, it is worth underlying how pro-inflammatory cytokines appear to be increased in blood or brain of patients with major depression (MD) and that pharmacological use of pro-inflammatory cytokines (i.e. interferon alpha) may induce MD. These data suggest a role for inflammation in the pathogenesis of depression and that anti-inflammatory drugs may be used as an adjunctive therapy in the treatment of MD. However, some studies reported contradictory results and suggest that adverse effects may contraindicate the use of anti-inflammatory agents in the treatment of depression. Nevertheless, non-steroidal anti-inflammatory drugs (NSAIDs) can have different mechanisms of action also depending on the dose. This is true for the therapeutic effects as well as for the unwanted side effects. On this basis, the present study aimed at evaluating the behavioural effect of the co-administration of fluoxetine (FLX, 5 mg/kg i.p.) with different NSAIDs in the chronic escape model of depression (CED). The CED model of depression possesses face, construct and pharmacological validity and is based on the induction, and maintenance, of an escape deficit upon exposing rats to unavoidable stressors. We previously demonstrated that, in this model, the stress-induced impaired behaviour can be resolved by one week of treatment with the co-administration of FLX (5 mg/kg/i.p.) plus acetylsalicylic acid (ASA, 45 mg/kg i.p.) but not FLX alone [1]. Here we evaluated the effect of the co-administration of FLX and flurbiprofen (FLB, an inhibitor of Cox-1 and Cox-2, 5 mg/kg, p.o.) or celecoxib (CLX, a selective COX2 inhibitor, 5 mg/kg, p.o.) in the CED model after 7 days of treatment. The co-administration FLX plus ASA (45mg/kg i.p.) was used as a positive control. Morever we tested the behavioral effect of different doses (45, 22.5 and 11.25 mg/Kg i.p.) of ASA as potentiating agent of the effect of fluoxetine. Our study shows that only the co-administration of ASA with FLX reverted the stress-induced condition of escape deficit after 7 days of treatment. Moreover, the amplitude of the antidepressant-like effect was dose dependent. The percentage of the antidepressant response was about 90%, 60% and 40% for animals receiving FLX (5 mg/kg/i.p.) plus ASA at the dose of 45, 22.5 or 11.25 mg/Kg i.p. respectively. Both flurbiprofen and celecoxib, when administered together with FLX for 7 days, failed to induce an antidepressant-like effects in the CED model. Higher dose of FLB (50 mg/Kg p.o.) and CLX (20 mg/Kg p.o.) were also tested, but they were associated to high mortality rate (80% and 25% respectively). These data demonstrated that neither all NSAIDs, nor all doses, may be useful in the treatment of depression while adverse effects can be potentiated or induced by the co-administration with antidepressants. Unraveling the cellular and molecular mechanisms behind the dissimilar behavioral response elicited by different anti-inflammatory drugs can contribute to understand the role of inflammation in the etiopathogensis of MD and to improve patient care. [1] Brunello, N., Alboni, S., Capone, G., Benatti, C., Blom, J.M., Tascedda, F., Kriwin, P., Mendlewicz, J., 2006 Shortened onset of action of antidepressants in major depression using acetylsalicylic acid augmentation: a pilot open-label study. Int Clin Psychopharmacol. 21:227-31.

2015 - The HD project" - biomarkers of depression in HIV newly diagnosed patients [Abstract in Rivista]
Ferrari, Silvia; Menozzi, Marianna; Alboni, Silvia; Miselli, Manuela; Rigatelli, Marco; Ghidoni, Andrea
abstract

Background: Inflammatory mediators may be relevant to explain the frequent comorbidity between depression, neurocognitive disorders and HIV. HIV induces activation of inflammatory mediators, mainly cytokines, that have been involved in the onset of depression and response to antidepressant treatment. Aim: To identify recurring profiles of inflammatory biomarkers subtending depression, effectiveness of antidepressants and neurocognitive disorders among HIV-infected individuals. Method: All adult newly HIV-diagnosed out-patients attending HIV clinics in three towns of Northern Italy will be screened, assessed for depression and studied immunologically and for neurocognitive disorders. The study-specific set of immune biomarkers will consist of: IL-1β and α, IL-6, IL-23, IL-18, IFN-γ, TNFα, MCP1, IL-8, BDNF, FGF-8, CNTF; IL1Ra, IL18Rα type II, IL18Rβ short, IL18BP. Results: 17 patients have been enrolled so far: of these, 6 (35%) were positive to PHQ-9 screening. Three of the 6 PHQ-9-positive patients were positive to the diagnostic assessment for depression. No neurocognitive disorders were found among the 17 patients. As the project will develop, it is expected that frequency of depression, neurocognitive disorders and effective antidepressant treatment will be found to correlate to the profile of immune biomarkers. These findings might help to understand the etiology of depression in HIV, and specifically the role of inflammation and immunological changes.

2014 - Behavioural and transcriptional effects of escitalopram in the chronic escape deficit model of depression [Articolo su rivista]
Benatti, Cristina; Alboni, Silvia; Blom, Johanna Maria Catharina; Gandolfi, Francesco; Mendlewicz, Julien; Brunello, Nicoletta; Tascedda, Fabio
abstract

The study of depression is facing major challenges: first, the need to develop new drugs with a faster onset of action and second, fulfilling the unmet needs of treatment resistant patients with more effective compounds. The chronic escape deficit (CED) is a valid and useful model of depression and is based on the induction of an escape deficit after exposure of rats to an unavoidable stress. This behavioural model provides a method for evaluating the capacity of a treatment to revert the escape deficit. The majority of antidepressant drugs need to be administered for at least 3-4 weeks in order to revert the escape deficit. A 7-day treatment with escitalopram reverted the stress-induced escape deficit in approximately 50% of the animals. Escitalopram treatment decreased anxiety-related behaviours in stressed animals, by increasing the time spent in the central part of the arena with respect to saline treated stressed animals, without affecting exploratory related behaviours. Gene expression profiling was carried out in the hippocampus to identify new targets associated with the effects of stress or with the different response to escitalopram. By combining a well-validated animal model with gene expression analysis we demonstrated that the CED model may represent a perfect tool for studying treatment-resistant depression.

2014 - Interleukin 18 activates MAPKs and STAT3 but not NF-κB in hippocampal HT-22 cells [Articolo su rivista]
Alboni, Silvia; Montanari, C; Benatti, Cristina; Sanchez Alavez, M; Rigillo, Giovanna; Blom, Johanna Maria Catharina; Brunello, Nicoletta; Conti, B; Pariante, Mc; Tascedda, Fabio
abstract

Interleukin (IL)-18 is a cytokine previously demonstrated to participate in neuroinflammatory processes. Since the components of the IL-18 receptor complex are expressed in neurons throughout the brain, IL-18 is also believed to directly influence neuronal function. Here we tested this hypothesis on mouse hippocampal neurons by measuring the effects of IL-18 on three pathways previously shown to be regulated by this cytokine in non-neuronal cells: the MAPK pathways, p38 and ERK1/2 MAPKs, STAT3 and NF-κB. Experiments were carried out in vitro using the immortalized hippocampal neuronal line HT-22 or in vivo following i.c.v. injection with recombinant mouse IL-18. We showed that IL-18 did not activate NF-κB in HT-22 cells whereas it induced a rapid (within 15min) activation of the MAPK pathways. Moreover, we demonstrated that IL-18 treatment enhanced P-STAT3 (Tyr705)/STAT3 ratio in the nucleus of HT-22 cells after 30-60min of exposure. A similar increase in P-STAT3 (Tyr705)/STAT3 ratio was observed in the whole hippocampus one hour after i.c.v. injection. These data demonstrate that IL-18 can act directly on neuronal cells affecting the STAT3 pathway; therefore, possibly regulating the expression of specific genes within the hippocampus. This effect may help to explain some of the IL-18-induced effects on synaptic plasticity and functionality within the hippocampal system.

2014 - Successful treatment of HIV-1 infection increases the expression of a novel, short transcript for IL-18 receptor α chain [Articolo su rivista]
Nasi, Milena; Alboni, Silvia; Pinti, Marcello; Tascedda, Fabio; Benatti, Cristina; Benatti, Stefania; Gibellini, Lara; DE BIASI, Sara; Borghi, Vanni; Brunello, Nicoletta; Mussini, Cristina; Cossarizza, Andrea
abstract

The importance of interleukin (IL)-18 in mediating immune activation during HIV infection has recently emerged. IL-18 activity is regulated by its receptor (IL-18R), formed by an α and a β chain, the IL-18-binding protein, and the newly identified shorter isoforms of both IL-18R chains. We evaluated gene expression of the IL-18/IL-18R system in peripheral blood mononuclear cells from HIV+ patients. Compared with healthy donors, IL-18 expression decreased in patients with primary infection. The IL-18Rα short transcript expression was strongly upregulated by successful highly active antiretroviral therapy. HIV progression and its treatment can influence the expression of different components of the complex IL-18/IL-18R system.

2014 - The role of microglia in mediating the effect of the environment in brain plasticity and behavior. [Articolo su rivista]
Branchi, I; Alboni, Silvia; Maggi, L.
abstract

No abstract available

2013 - Chronic antidepressant treatments resulted in altered expression of genes involved in inflammation in the rat hypothalamus [Articolo su rivista]
Alboni, Silvia; Benatti, Cristina; Montanari, Claudia; Tascedda, Fabio; Brunello, Nicoletta
abstract

To gain insight into the possible immune targets of antidepressant, we evaluated the expression of several inflammatory mediators in the hypothalamus of rats chronically (28 days) treated with the serotonin selective reuptake inhibitor fluoxetine (5mg/kg, i.p.) or the tricyclic compound imipramine (15 mg/kg, i.p.). We focused our attention on the hypothalamus as it plays a key role in determining many of the somatic symptoms experienced by depressed patients. This brain region, critical also for expression of motivated behaviours, participates in the control of the hypothalamic-pituitary-adrenal axis activity and in stress response as well as coordinates physiological functions such as sleep and food intake that have been found altered in a high percentage of depressed patients. Notably, hypothalamus is a key structure for brain cytokine expression and function as it integrates signals from the neuro, immune, endocrine systems. By means of quantitative Real Time PCR experiments we demonstrated that a chronic treatment with either fluoxetine or imipramine resulted in a reduction of IL-6 and IFN-γ mRNAs and increased IL-4 mRNA expression in the rat hypothalamus. Moreover, we demonstrated that hypothalamic expression of members of IL-18 system was differentially affected by chronic antidepressant treatments. Chronically administered fluoxetine decreased IL-8 and CX3CL1 hypothalamic expression, while a chronic treatment with imipramine decreased p11 mRNA. Our data suggest that a shift in the balance of the inflammation toward an anti-inflammatory state in the hypothalamus may represent a common mechanism of action of both the chronic treatments with fluoxetine and imipramine.

2013 - Interferon alpha exposure increases the expression of the enzymes belonging to the kynurenine pathway in an in vitro model of human neurons: SH-SY5Y cells [Abstract in Rivista]
Alboni, Silvia; Benatti, Cristina; Claudia, Montanari; Benatti, Stefania; Tascedda, Fabio; Cannazza, Giuseppe; Pariante Carmine, M; Brunello, Nicoletta
abstract

The past two decades have witnessed a burgeoning area of pre-clinical and clinical research linking psychiatric illnesses – particularly major depression (MD) – to activation of the inflammatory immune system. One of the stronger evidence supporting a causal role for inflammation in leading MD comes from reports indicating that depressive symptoms frequently develop in patients undergoing immunotherapy with cytokines, such as interferon (IFN)-α, for the treatment of malignancies or chronic viral infection. Although INF-alpha- induced effects on the brain made of IFN-α a model to study the influence of pro-inflammatory cytokines in the CNS and behavior the molecular mechanisms underlying these effects are far from being fully understood. It has been proposed that IFN-α may contribute to the etiology of MD by inducing indolamine 2,3-dioxygenase (IDO) expression and thus unbalancing in the tryptophan/kynurenine metabolism toward the production of neurotoxic metabolites and\or reducing serotonin (5-HT) availability. IDO catalyzes the initial rate-limiting step in tryptophan degradation along the kynurenine pathway (KP). Kynurenine, the initial product of tryptophan degradation, is further catalysed into neurotoxic end-products through steps catalyzed by kynurenine 3-monooxygenase (KMO) and kynureninase (Kynu). However, Kynurenine can also be catabolised by kynurenine aminotransferase (KAT), into kynurenic acid, a potentially neuroptotective agent. A role for a disturbance in the equilibrium between neurotoxic/ neuropoptective end KP endproducts producing an alteration in the neuroprotective–neurodegenerative balance in the brain of patients with MD, has been proposed in the neurodegeneration hypothesis of depression. Given that we previously demonstrated that IFN-α induces toxic effects in an in vitro model of human neurons (human SH-SY5Y neuroblastoma cells) we were aim to investigate the effects of IFN-α on KP in these cells. Our studies show that IFN-α exposure increased the expression of all the kynurenergic enzymes investigated (IDO, KMO, Kynu and KAT). More particularly strongly induced the expression of IDO mRNA (more than 900 –fold) in SH-SY5Y cells. Similar effects on kynurenergic enzyme expression were also observed when SH-SY5Y cells where differentiated with all-trans retinoic acid (in presence of neurotrophic support and in serum deprived conditions). We also demonstrated that INF-α decreased 5-HT levels whereas increased the kynurenine levels in the medium of both differentiated as well not differentiated SH-SY5Y cells.

2013 - Metabolic changes induced by interferon-α exposure in an in vitro model of human neurons [Abstract in Rivista]
Alboni, Silvia; Schenetti, Luisa; Brunello, Nicoletta; Pariante Carmine, M; Righi, Valeria
abstract

The human neuroblastoma SH-SY5Y cell line is a third successive subclone of the SK-N-SH line, originally established from a bone marrow biopsy of a neuroblastoma patient. These cells possess many characteristics of neurons, and they represent one of the most-used models for studying cellular events and mechanisms involved in neurotoxicity and neurodegeneration or even in neuroprotection. We have been using these cells as a tools to evaluate the largely unexplored effects induced by interferon (IFN)- alpha (a clinically used type I IFN) exposure on neurons. Interferons are cytokines endowed with a pleiotropic spectrum of biological properties, including immunomodulation, antiviral and pro-inflammatory activity. Beside the periphery, and cells of the immune system, type I IFNs may have broad-ranging actions also in the brain, affecting neuronal differentiation, survival and synaptic plasticity. We recently demonstrated that exposure to IFN-α induces neurotoxic effects in a time e dose dependent manner in SH-SY5Y cells by impairing mitochondrial integrity and activity, recruitment of Bcl-2 family members, induction of oxidative stress (increases reactive oxygen species). Indeed following 72 hours exposure to this cytokine we found increased early apoptosis (Alboni et al., 2013). This prompted us to investigate the changes in the metabolic profile of live SH-SY5Y cells exposed to IFN-α (72 h) using 1H High Resolution-Magic Angle Spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy. Firstly, this technique enabled us to characterize the metabolic signature of intact SH-SY5Y cells. Several metabolites, including amino acids, osmolites, phospholipids, organic acids, sugars and polyols have been identified. Moreover, we found that human neuronal cells exposed to IFN-α for 72 h had significantly increased concentration of lactate (% 93.9), taurine (% 117.7), myo-inositol (% 123.2), scyllo-inositol (% 117.4), glycerolphosphocholine (% 135.2) and creatine (% 159.9) compared with the vehicle-treated control cells. These data provide the demonstration that IFN-α exposure induces metabolic changes in human neuron-like cells. Moreover, these results may contribute to explain IFN-α-induced central side-effects often observed following IFN-α treatment for viral infection and malignancies.

2013 - Metabolic characterization of SH-SY5Y cells and effects of interferon-alpha exposure [Abstract in Atti di Convegno]
Alboni, Silvia; Schenetti, Luisa; Mucci, Adele; Brunello, Nicoletta; Valeria, Righi
abstract

The human neuroblastoma SH-SY5Y cell line is a third successive subclone of the SK-N-SH line, originally established from a bone marrow biopsy of a neuroblastoma patient. These cells possess many characteristics of neurons, and they represent one of the most-used models for studying cellular events and mechanisms involved in neurotoxicity and neurodegeneration or even in neuroprotection. Interferons are cytokines endowed with a pleiotropic spectrum of biological properties, including immunomodulation, antiviral and proinflammatory activity. Interferon (IFN)-α is a type I IFN that may have broad-ranging actions in the brain, affecting neuronal differentiation, survival and synaptic plasticity. We previously demonstrated that a 72 hours exposure to IFN-α induces early apoptosis in SH-SY5Y cells. This prompted us to investigate the metabolic profile of the SH-SY5Y cells using HR-MAS NMR Spectroscopy after a 72 hours exposure to IFN-α to explore the metabolic changes that characterize these cells. Moreover, since a metabolic characterization of this extensively used cell clone is still lacking, we analyzed the metabolic profile of the SH-SY5Y in standard growth conditions. Results show some interesting changes in metabolites, such as choline containing compounds, creatine and glutamate. Our goal will be to relate the metabolic changes to IFN-α exposure.

2013 - N-acetyl-cysteine prevents toxic oxidative effects induced by IFN-α in human neurons. [Articolo su rivista]
Alboni, Silvia; Gibellini, Lara; Montanari, Claudia; Benatti, Cristina; Benatti, Stefania; Tascedda, Fabio; Brunello, Nicoletta; Cossarizza, Andrea; Pariante, Carmine M.
abstract

Currently IFN-α is widely used for effective treatment of viral infections and several malignancies. However, IFN-α can cause neuropsychiatric disturbances and mental impairments, including fatigue, insomnia, depression, irritability and cognitive deficits. Molecular and cellular mechanisms leading to such side-effects are still poorly understood. Neurons seem to be an important target in mediating cellular effects induced by exposure to this cytokine, but so far little is known about IFN-α-induced effects on these cells. We have investigated the ability of IFN-α (2-100 ng/ml) to induce damage and toxicity to the human neuroblastoma SH-SY5Y cell line, commonly used for studying such phenomena, and the mechanisms underlying these effects. After 24 h treatment, IFN-α increased mitochondrial activity, whereas cell density was reduced in a dose- and time-dependent manner. This effect did not depend on reduced cell proliferation, but rather the activation of apoptosis, as revealed by an increased Bax:Bcl-2 mRNA ratio after 72-h IFN-α exposure. At this time-point, IFN-α also reduced the expression of the brain-derived neurotrophic factor gene, and induced an increase in reactive oxygen species (ROS). A co-treatment with N-acetyl-cysteine (NAC; 5 mm), a potent antioxidant and mitochondrial modulator, was able to counteract all of these IFN-α-induced effects. These findings demonstrated that IFN-α induces neurotoxicity and apoptosis that is, in part, very likely due to mitochondrial damages and production of ROS. We suggest that NAC, already tested for the treatment of psychiatric disorders, may be useful to prevent IFN-α-induced central side-effects in a safe and effective way.

2013 - Neuropeptide S stimulates human monocyte chemotaxis via NPS receptor activation. [Articolo su rivista]
Filaferro, Monica; Novi, Chiara; Ruggieri, Valentina; Genedani, Susanna; Alboni, Silvia; Malagoli, Davide; Caló, G; Guerrini, R; Vitale, Giovanni
abstract

Neuropeptide S (NPS) produces several biological actions by activating a formerly orphan GPCR, now named NPS receptor (NPSR). It has been previously demonstrated that NPS stimulates murine leukocyte chemotaxis in vitro. In the present study we investigated the ability of NPS, in comparison with the proinflammatory peptide formyl-Met-Leu-Phe (fMLP), to stimulate human monocyte chemotaxis. At a concentration of 10(-8)M fMLP significantly stimulated chemotaxis. NPS produced a concentration dependent chemotactic action over the concentration range 10(-12) to 10(-5)M. The NPSR antagonists [D-Cys((t)Bu)(5)]NPS, [(t)Bu-D-Gly(5)]NPS and SHA 68 were used to pharmacologically characterize NPS action. Monocyte chemoattractant effect of NPS, but not fMLP, was completely blocked by either peptide antagonists or SHA with the nonpeptide molecule being more potent. None of the NPSR antagonists modified per se random cell migration. Thus, the present study demonstrated that NPS is able to stimulate human monocyte chemotaxis and that this effect is entirely due to selective NPSR activation.

2012 - Interferon-alpha exposure increases the expression of enzymes of the kynurenine pathway and induces apoptosis in a model of human neurons [Abstract in Rivista]
Alboni, Silvia; Benatti, Cristina; Montanari, Claudia; Tascedda, Fabio; C. M., Pariante; Brunello, Nicoletta
abstract

Major depression (MD) is associated with a profound unbalance between the nervous-, the endocrine- and the immune- systems. This suggests the possibility that molecules that regulate the homeostasis of these systems may contribute to the development of MD. For instance, some cytokines, important neuro-endocrineimmuno modulators, have been proposed to have a role in MD as supported by the observation that activation of immune system with therapeutically used cytokines may induce MD. IFN-a is an innate immune cytokine with potent antiviral and anti-proliferative properties that is used to treat viral infections, such as hepatitis C, and certain cancers. Despite therapeutic efficacy in these illnesses, it has been observed that IFN-a exposure may be associated with important side effects including neuropsychological and behavioural changes that overlap with MD. Although IFN-a- induced effects on the brain make IFN-alpha a model to study the influence of pro-inflammatory cytokines in the CNS and behavior, the molecular mechanisms underlying these effects are far from being fully understood. It has been proposed that cytokines may contribute to the etiology of MD by inducing indolamine 2,3-dioxygenase (IDO) expression. IDO catalyzes the initial rate-limiting step in tryptophan (TPR) degradation along the kynurenine pathway (KP). Kynurenine, the initial product of TPR degradation, is further catalysed into neurotoxic end-products through steps catalyzed by kynurenine 3-monooxygenase (KMO) and kynureninase (Kynu). However, Kynurenine can also be catabolised by kynurenine aminotransferase (KAT), into kynurenic acid, a potentially neuroprotective agent. A role for a disturbance in the KP in the neuroprotective–neurodegenerative balance in the brain of patients with MD, has been proposed in the neurodegeneration hypothesis of depression. This prompted us to investigate the effects of IFN-a on the expression of the IDO, KMO, Kynu and KAT mRNAs in an in vitro model of human neurons: SH-SY5Y (human neuroblastoma) cells. We also evaluated the IFN-a-induced effect on cell viability and number in these cells. Finally, because cell number results from the balance between cell proliferation and cell elimination, we measured cell proliferation and apoptosis in SH-SY5Y cells after IFN-a exposure. The same experiments were performed in differentiated SH-SY5Y cells with retinoic acid (RA) and Brain- Derived Neurotrophic Factor (BDNF). Our studies show that IFN-a exposure increased the expression of all the kynurenergic enzymes investigated (with an unbalance of the KP toward the synthesis of neurotoxic end-products) and in more particularly strongly induced the expression of IDO mRNA (more than 900-fold) in SH-SY5Y cells. We also demonstrated that IFN-a reduced in a dose- (2, 20, 50 and 100 ng/ml) and time- (24, 48 and 72 hrs) dependent manner the cell number and induces apoptosis in SH-SY5Y cells. Similar results were obtained in SHSY5Y RA/BDNF differentiated cells. Together our results clearly enlighten the cytotoxic effects of IFN-a in this in vitro model of human neurons. Moreover, our findings provide further information on the molecular pathways involved in cytokine-induced effects in the brain and add a piece to the puzzle of what and how these factors or pathways may contribute to the pathogenesis of MD.

2012 - Transcriptional profiles underlying vulnerability and resilience in rats exposed to an acute unavoidable stress [Articolo su rivista]
Benatti, Cristina; Valensisi, Cristina; Blom, Johanna Maria Catharina; Alboni, Silvia; Montanari, Claudia; Ferrari, Francesco; Tagliafico, Enrico; Julien, Mendlewicz; Brunello, Nicoletta; Tascedda, Fabio
abstract

A complex interplay between gene and environment influences the vulnerability or the resilience to stressful events. In the acute escape deficit (AED) paradigm, rats exposed to an acute unavoidable stress (AUS) develop impaired reactivity to noxious stimuli. Here we assessed the behavioral and molecular changes in rats exposed to AUS. A genome-wide microarray experiment generated a comprehensive picture of changes in gene expression in the hippocampus and the frontal cortex of animals exposed or not to AUS. Exposure to AUS resulted in two distinct groups of rats with opposite behavioral profiles: one developing an AED, called “stress vulnerable,” and one that did not develop an AED, called “stress resilient.” Genome-wide profiling revealed a low percentage of overlapping mechanisms in the two areas, suggesting that, in the presence of stress, resilience or vulnerability to AUS is sustained by specific changes in gene expression that can either buffer or promote the behavioral and molecular adverse consequences of stress. Specifically, we observed in the frontal cortex a downregulation of the transcript coding for interferon-β and leukemia inhibitory factor in resilient rats and an upregulation of neuroendocrine related genes, growth hormone and prolactin, in vulnerable rats. In the hippocampus, the muscarinic M2 receptor was downregulated in vulnerable but upregulated in resilient rats. Our findings demonstrate that opposite behavioral responses did not correspond to opposite regulatory changes of the same genes, but resilience rather than vulnerability to stress was associated with specific changes, with little overlap, in the expression of patterns of genes.

2011 - Central effects of a local inflammation in three commonly used mouse strains with a different anxious phenotype [Articolo su rivista]
Benatti, Cristina; Alboni, Silvia; Montanari, Claudia; Caggia, Federica; Tascedda, Fabio; Brunello, Nicoletta; Blom, Johanna Maria Catharina
abstract

As in humans, genetic background in rodents may influence a peculiar set of behavioural traits such as sensitivity to pain and stressors or anxiety-related behaviours. Therefore, we tested the hypothesis that mice with different genetic backgrounds [outbred (CD1), inbred (C57BL/6J) and hybrid (B6C3F1) adult male mice] display altered reactivity to pain, stress and anxiety related behaviours. We demonstrated that B6C3F1 mice displayed the more anxious phenotype with respect to C57BL/6J or CD1 animals, with the latter being the less anxious strain when tested in an open field and on an elevated plus maze. No difference was observed across strains in thermal sensitivity to a radiant heat source. Mice were then treated with a sub-plantar injection of the inflammatory agent Complete Freund's Adjuvant (CFA), 24h later they were hyperalgesic with respect to saline exposed animals, irrespective of strain. We then measured intra-strain differences and CFA-induced inter-strain effects on the expression of various genes with a recognized role in pain and anxiety: BDNF, IL-6, IL-1β, IL-18 and NMDA receptor subunits in the mouse thalamus, hippocampus and hypothalamus. The more anxious phenotype observed in B6C3F1 hybrid mice displayed lower levels of BDNF mRNA in the hippocampus and hypothalamus when compared to outbred CD1 and C57BL/6J inbred mice. CFA led to a general decrease in central gene expression of the evaluated targets especially in CD1 mice, while BDNF hypothalamic downregulation stands out as a common effect of CFA in all three strains evaluated

2011 - Constitutive and LPS-regulated expression of interleukin-18 receptor beta variants in the mouse brain [Articolo su rivista]
Alboni, Silvia; Montanari, Claudia; Benatti, Cristina; Blom, Johanna Maria Catharina; Simone, Ml; Brunello, Nicoletta; Caggia, Federica; Guidotti, G; Marcondes, Mc; Sanchez Alavez, M; Conti, B; Tascedda, Fabio
abstract

Interleukin (IL)-18 is a pro-inflammatory cytokine that is proposed to be involved in physiological as well as pathological conditions in the adult brain. IL-18 acts through a heterodimer receptor comprised of a subunit alpha (IL-18Rα) required for binding, and a subunit beta (IL-18Rβ) necessary for activation of signal transduction. We recently demonstrated that the canonical alpha binding chain, and its putative decoy isoform, are expressed in the mouse central nervous system (CNS) suggesting that IL-18 may act on the brain by directly binding its receptor. Considering that the co-expression of the beta chain seems to be required to generate a functional receptor and, a short variant of this chain has been described in rat and human brain, in this study we have extended our investigation to IL-18Rβ in mouse. Using a multi-methodological approach we found that: (1) a short splice variant of IL-18Rβ was expressed in the CNS even if at lower levels compared to the full-length IL-18Rβ variants, (2) the canonical IL-18Rβ is expressed in the CNS particularly in areas and nuclei belonging to the limbic system as previously observed for IL-18Rα and finally (3) we have also demonstrated that both IL-18Rβ isoforms are up-regulated in different brain areas three hours after a single lipopolysaccharide (LPS) injection suggesting that IL-18Rβ in the CNS might be involved in mediating the endocrine and behavioral effects of LPS. Our data highlight the considerable complexity of the IL-18 regulation activity in the mouse brain and further support an important central role for IL-18.

2011 - Sex- and age-specific differences in core body temperature of C57Bl/6 mice [Articolo su rivista]
Sanchez Alavez, M; Alboni, Silvia; Conti, B.
abstract

Gender-specific differences in longevity are reported across species and are mediated by mechanisms not entirely understood. In C57Bl/6 mice, commonly used in aging research, males typically outlive females. Since in these animals modest but prolonged reduction of core body (Tc) increased life span, we hypothesized that differential Tc may contribute to sex-specific longevity. Here, we compared the circadian profiles of Tc and locomotor activity (LMA) of male and female C57Bl/6 mice. Since Tc and LMA normally change with age, measurements were carried out in young (3 months) as well as in old (24 months) mice. In young females, Tc was influenced by estrous but was overall higher than in males. This difference was larger in old animals after age eliminated the variations associated with estrous. Although temperature homeostasis is regulated centrally by the sexually dimorphic hypothalamic preoptic area, these differences were uniquely dependent on the gonads. In fact, bilateral gonadectomy abolished the effects of estrous and increased resting Tc in males eliminating all sex-specific differences in Tc and LMA. These effects were only partially mimicked by hormonal replacement as Tc was affected by progesterone and to a lesser extent by estrogen but not by testosterone. Thus, gonadal-dependent modulation of Tc may be one of the physiological parameters contributing to gender-specific differences in longevity.

2011 - Stress induces altered CRE/CREB pathway activity and BDNF expression in the hippocampus of glucocorticoid receptor-impaired mice [Articolo su rivista]
Alboni, Silvia; Tascedda, Fabio; Corsini, Daniela; Benatti, Cristina; Caggia, Federica; Capone, Giacomo; Barden, N; Blom, Johanna Maria Catharina; Brunello, Nicoletta
abstract

The gene coding for the neurotrophin Brain-Derived Neurotrophic Factor (BDNF) is a stress-responsive gene. Changes in its expression may underlie some of the pathological effects of stress-related disorders like depression. Data on the stress-induced regulation of the expression of BDNF in pathological conditions are rare because often research is conducted using healthy animals. In our experiments, we used transgenic mice with glucocorticoid receptor impaired (GR-i) expression in the hypothalamus created as a tool to study the neuroendocrine changes occurring in stress-related disorders. First, under basal condition, GR-i mice displayed lower levels of BDNF exons IX and IV and decreased CRE(BDNF) binding activity with respect to wild-type (WT) mice in the hippocampus. Then, we exposed GR-i and WT mice to an acute restraint stress (ARS) to test the hypothesis that GR-i mice display: 1] different ARS induced expression of BDNF, and 2] altered activation of signaling pathways implicated in regulating BDNF gene expression in the hippocampus with respect to WT mice. Results indicate that ARS enhanced BDNF mRNA expression mainly in the CA3 hippocampal sub-region of GR-i mice in the presence of enhanced levels of pro-BDNF protein, while no effect was observed in WT mice. Moreover, ARS reduced CREB signaling and binding to the BDNF promoter in GR-i mice but enhanced signaling and binding, possibly through ERK1/2 activation, in WT mice. Thus, life-long central GR dysfunction resulted in an altered sensitivity at the transcriptional level that may underlie an impaired response to an acute psycho-physical stress.

2010 - Erratum: HTR1B as a risk profile maker in psychiatric disorders: A review through motivation and memory (European Journal of Clinical Pharmacology DOI:10.1007/s00228-009-0724-6) [Articolo su rivista]
Drago, A.; Alboni, S.; Brunello, N.; De Ronchi, D.; Serretti, A.
abstract

2010 - HTR1B as a risk profile maker in psychiatric disorders:a review through motivation and memory [Articolo su rivista]
Antonio, Drago; Alboni, Silvia; Brunello, Nicoletta; Diana De, Ronchi; Alessandro, Serretti
abstract

Purpose Serotonin receptor 1B (HTR1B) is involved in theregulation of the serotonin system, playing different roles inspecific areas of the brain. We review the characteristics ofthe gene coding for HTR1B, its product and the functionalrole of HTR1B in the neural networks involved inmotivation and memory; the central role played by HTR1Bin these functions is thoroughly depicted and show HTR1Bto be a candidate modulator of the mnemonic andmotivationally related symptoms in psychiatric illnesses.Methods In order to challenge this assessment, we analyzehow and how much the genetic variations located in thegene that codes for HTR1B impacts on the psychiatricphenotypes by reviewing the literature on this topic.Results We gathered partial evidence arising from geneticassociation studies, which suggests that HTR1B plays arelevant role in substance-related and obsessive compulsivedisorders. On the other hand, no solid evidence for otherpsychiatric disorders was found. This finding is quitestriking because of the heavy impairment of motivationand of mnemonic-related functions (for example, recallbias) that characterize major psychiatric disorders.Conclusions The possible reasons for the contrast betweenthe prime relevance of HTR1B in regulating memory andmotivation and the limited evidence brought by geneticassociation studies in humans are discussed, and somesuggestions for possible future directions are provided.

2010 - Interleukin 18 in the CNS [Articolo su rivista]
Alboni, Silvia; Cervia, D; Sugama, S; Conti, B.
abstract

Interleukin (IL)-18 is a cytokine isolated as an important modulator of immune responses and subsequently shown to be pleiotropic. IL-18 and its receptors are expressed in the central nervous system (CNS) where they participate in neuroinflammatory/neurodegenerative processes but also influence homeostasis and behavior. Work on IL-18 null mice, the localization of the IL-18 receptor complex in neurons and the neuronal expression of decoy isoforms of the receptor subunits are beginning to reveal the complexity and the significance of the IL-18 system in the CNS. This review summarizes current knowledge on the central role of IL-18 in health and disease.

2010 - Time-dependent effects of escitalopram on brain derived neurotrophic factor (BDNF) and neuroplasticity related targets in the central nervous system of rats [Articolo su rivista]
Alboni, Silvia; Benatti, Cristina; Capone, Giacomo; Corsini, Daniela; Caggia, Federica; Tascedda, Fabio; Mendlewicz, J; Brunello, Nicoletta
abstract

Chronic treatment with antidepressants affects several proteins linked to neuroplasticity, particularly brain derived neurotrophic factor (BDNF): this leads eventually to their therapeutic effects. It is possible that also for putative early therapeutic onset, antidepressants may act by promoting cellular adaptations linked to neuroplasticity. Escitalopram, known to be already effective in preclinical models of depression after 7 days, allowed us to investigate whether two effective treatment regimens (7 and 21 days) may contribute to synaptic plasticity by acting on BDNF signalling. We focused our attention on two regulators of BDNF transcription, CREB and CaRF (calcium responsive factor), and on kinases, CaMKII, ERK1/2 and p38 MAPK, linked to BDNF that play a distinctive role in synaptic plasticity. We evaluated whether the effects of escitalopram on these targets may be different in brain areas involved in the depressive symptomatology (hippocampus, frontal and prefrontal cortex). Here we demonstrate that escitalopram regulates intracellular pathways linked to neuroplasticity at both the time points evaluated in an area-specific manner. While the two escitalopram-treatment regimens failed to affect gene expression in the rat frontal cortex, 7days of treatment with escitalopram activated intracellular pathways linked to BDNF and increased the levels of Pro-BDNF in the rat prefrontal cortex. Moreover, 21 days of treatment with escitalopram decreased CREB/BDNF signalling while increasing p38 levels in the rat hippocampus. Even if further experiments with different antidepressant strategies will be needed, our data suggest that escitalopram efficacy may be mediated by early and late effects on synaptic plasticity in selective brain areas.

2009 - Chronic treatment with the selective NOP receptor antagonist [Nphe1,Arg14,Lys15]N/OFQ-NH2 (UFP-101) reverses the behavioural and biochemical effects of unpredictable chronic mild stress in rats [Articolo su rivista]
Vitale, Giovanni; Ruggieri, Valentina; Filaferro, Monica; Frigeri, Claudio; Alboni, Silvia; Tascedda, Fabio; Brunello, Nicoletta; Guerrini, R; Cifani, C; Massi, M.
abstract

Introduction The present study was designed to assess the antidepressant effects of UFP-101, a selective nociceptin/orphanin FQ peptide (NOP) receptor antagonist, in a validated animal model of depression: the chronic mild stress (CMS). Materials and methods and Results UFP-101 (5, 10 and 20 nmol/rat; i.c.v., once a day for 21 days) dose- and time-dependently reinstated sucrose consumption in stressed animals without affecting the same parameter in non-stressed ones. In the forced swimming test, UFP-101 reduced immobility of stressed rats from day 8 of treatment. After a 3-week treatment, rats were killed for biochemical evaluations. UFP-101 abolished increase in serum corticosterone induced by CMS and reverted changes in central 5-HT/5-HIAA ratio. The behavioural and biochemical effects of UFP-101 mimicked those of imipramine, the reference antidepressant drug, administered at the dose of 15 mg/kg (i.p.). Co-administration of nociceptin/orphanin FQ (5 nmol/rat, from day 12 to 21) prevented the effects of UFP-101. Brain-derived neurotrophic factor mRNA and protein in hippocampus were not reduced by CMS nor did UFP-101 modify these parameters. Discussion and Conclusion This study demonstrated that chronic treatment with UFP-101 produces antidepressant-like effects in rats subjected to CMS supporting the proposal that NOP receptors represent a candidate target for the development of innovative antidepressant drugs.

2009 - Early neonatal inflammation affects adult pain reactivity and anxiety related traits in mice: genetic background counts [Articolo su rivista]
Benatti, Cristina; Alboni, Silvia; Capone, Giacomo; Corsini, Daniela; Caggia, Federica; Brunello, Nicoletta; Tascedda, Fabio; Blom, Johanna Maria Catharina
abstract

Protracted or recurrent pain and inflammation in the early neonatal period may cause long-lasting changes in central neural function. However, more research is necessary to better characterize the long-term behavioral sequelae of such exposure in the neonatal period. Objectives: (1) to study whether timing of postnatal exposure to persistent inflammation alters responsiveness to thermal pain in the adult animal; (2) to assess whether animals experiencing early postnatal chronic inflammation display altered anxiety related behavior; (3) to study the importance of genetic background. Newborn mice (outbred strain, CD1 and F1 hybrid strain, B6C3F1) received an injection of Complete Freund's Adjuvant (CFA) or saline on either postnatal day 1 or 14 (PND1; PND14) into the left hind paw. Pain to radiant heat and anxiety were examined in 12-week-old adult animals. Adult baseline PWL was significantly decreased in CD1 mice exposed to CFA on PND 1 and 14 as compared to their saline treated counterparts. B6C3F1 mice exposed to CFA on PND14 showed markedly reduced baseline PWL compared to the PND14 saline group. Persistent inflammation experienced by B6C3F1 mice on PND1 failed to affect baseline adult thermal responsiveness. Adult mice, CD1 and B6C3F1, displayed low anxiety traits only if they had been exposed to persistent inflammation on PND1 and not on PND14. Our research suggests a role for genetic background in modulating long-term behavioral consequences of neonatal persistent inflammation: the data support the hypothesis that pain experienced very early in life differentially affects adult behavioral and emotional responsiveness in outbred (CD1) and hybrid mice (B6C3F1).

2009 - Gene expression profile of the hippocampus of a behavioural model of depression [Abstract in Rivista]
Valensisi, Cristina; Caggia, Federica; Alboni, Silvia; Benatti, Cristina; Ferrari, F; Mendlewicz, J; Blom, Johanna Maria Catharina; Brunello, Nicoletta; Tascedda, Fabio
abstract

Although the neurobiological basis of depression has not been fully elucidated, numerous studies have emphasized that in the etiology of depression stress may be the most significant cause, together with genetic vulnerability. Stress induces a coordinated and complex response that is adaptive and integral to survival. The brain's ability to adapt and change over time is refered to neuroplasticity and long-term plasticity in the brain requires changes in gene expression. However, exposure to intense or chronic stressors leads to an increased risk for the development of stress-related disorders including major depression. Numerous studies demonstrate that neuronal atrophy and loss of plasticity occur in hippocampus in response to stress and depression. Therefore, the hippocampal region may play a central role in depressive illness. Likewise changes in gene expression underlying the plasticity of hippocampal structures appear to be relevant in undenstanding the molecular and cellular mechanisms involved in the etiology as well as the treatment of depression, and the mechanisms leading vulnerability or resilience to stress. In fact, humans display a remarkable heterogeneity in their responses to stress and adversity. Although we are beginning to understand how maladaptive neurobiological changes may contribute to depression, relatively little is known about the molecular mechanisms that may underlie stress resilience. Here we set out to investigate the changes in the gene expression profile underlying the effects of stress on the hippocampus using a behavioural paradigm of depression, the chronic escape deficit model [1], which is based on the modified reactivity of rats to external stimuli, the escape deficit, induced by exposure to intense and unavoidable stress. The chronic escape deficit model starts as an acute escape deficit which can be indefinitely sustained by repeated administration of mild stressors. This approach has proved to be a valid and useful model of depression because it consider depressive symptoms like behavioural despair. We performed gene expression profiling in the rat hippocampus, using GeneChip Rat Exon Array (Affymetrix). Using this new platform we carried out analyses of gene expression on three different levels: gene, transcript and exon level analyses. The behavioural results showed that exposure to intense and unavoidable stressful procedure induced escape deficit only in 60% of them. Whereas the animals remaining display a behaviour apparently identical to control animals which did not undergo the stressful procedure. Comparing gene expression profiles and performing functional analysis on differently expressed genes we have indicated multiple pathways that may be involved in the underlying mechanisms of stress condition associated with escape deficit. Moreover we identified possible cellular functions and biological processes that could represent targets that may contribute to mediate the effects of stress on the hippocampal plasticity. Such as, gene expression profiling of stress-vulnerable and stress-resilient animals revealed distinct transcriptional profiles, suggesting that resilient behaviour represents an active neurobiological process and not simply the absence of vulnerability.

2009 - Mapping of the full length and the truncated interleukin-18 receptor alpha in the mouse brain [Articolo su rivista]
Alboni, Silvia; Cervia, D; Ross, B; Montanari, Claudia; Gonzalez, As; Sanchez Alavez, M; Marcondes, Mc; De Vries, D; Sugama, S; Brunello, Nicoletta; Blom, Johanna Maria Catharina; Tascedda, Fabio; Conti, B.
abstract

The cytokine IL-18 acts on the CNS both in physiological and pathological conditions. Its action occurs through the heterodimeric receptor IL-18Rα β. To better understand IL-18 central effects, we investigated in the mouse brain the distribution of two IL-18Rα transcripts, a full length and an isoform lacking the intracellular domain hypothesized to be a decoy receptor. Both isoforms were expressed in neurons throughout the brain primarily with overlapping distribution but also with some unique pattern. These data suggest that IL-18 may modulate neuronal functions and that its action may be regulated through expression of a decoy receptor.

2009 - Microarray analysis in hippocampus of rats treated with escitalopram in the chronic escape deficit model of depression [Abstract in Rivista]
Caggia, Federica; Valensisi, Cristina; Alboni, Silvia; Benatti, Cristina; Corsini, Daniela; F., Ferrari; Tagliafico, Enrico; J., Mendlewicz; Tascedda, Fabio; Brunello, Nicoletta
abstract

Currently, the biological bases of depression and the molecular mechanisms underlying antidepressant action are not completely understood. Valuable tools to better understand the pathophysiology of this disease are behavioural models of depression eventually combined with genome-wide gene expression analysis. The Chronic Escape Deficit (CED) is a validated behavioural model of depression, based on the induction of an escape deficit after exposure of rats to an unavoidable stress. This model allows to evaluate the capacity of a treatment to revert the escape deficit. The antidepressant drugs tested in CED model need to be administered for at least 3−4 weeks in order to revert the escape deficit [1,2]. In this study, we demonstrated that already after one week of treatment with Escitalopram, a widely used SSRI, 50% of the animals responded reverting the escape deficit induced. Moreover, the other 50% of treated animals did not respond also after 3−4 weeks of treatment. Since in the CED model the behavioural alteration is induced by stress application and reverted by escitalopram treatment in only half of animals, the aims of our study were two fold: (i) to investigate transcriptional changes activated by stress; (ii) to study the different gene expression pattern involved into mechanisms of the response and not response to the pharmacological treatment. To address these issues we performed a microarray experiment in the rat hippocampus using Affymetrix GeneChip Rat Exon 1.0 ST evaluating both gene-level and exon-level expression profiling on the whole genome. Total RNA extracted from hippocampus of each animal was utilized to chip a single array using the Affymetrix protocols. 20 single arrays were utilized for data analysis and divided into five replicates for each experimental group (control, stress, stress-escitalopram responders and stress escitalopram-not responders). Using two parallel analyses (gene level and exon level) of raw data files carried out in Expression Console software using iterPLIER algorithms, we identified genes and exons that were differentially regulated in each pairwise comparison considered. The exons identified in this study were examinated for their biological association to gene ontology (GO) categories using eGOn software. Moreover, all exons differentially expressed were also uploaded into Ingenuity Pathways Analysis (Ingenuity® Systems, www.ingenuity.com) in order to identify molecular pathways and functions related to stress and escitalopram response. Our results suggest that stress may exert a negative effect on gene transcription since the largest number of genes was downregulated. Moreover from our data it seems that a different pattern of gene expression exhibits between animals that respond and that did not respond to escitalopram treatment. Functional analysis of exon dataset, arising from stress protocol and escitalopram treatment, reflects interesting different biological features. More specifically, the biological functions regard both molecular and cellular functions, such as cellular growth and proliferation, gene expression and signal transduction, as well as involvement of central neurotransmission and immune response. We believe that this pharmacogenomic approach will be helpful to understand the molecular mechanisms involved in the pathogenesis of depression as well as in the response to antidepressant drugs.

2008 - Impaired stress-induced regulation of brain derived neurotrophic factor expression in hippocampus of glucocorticoid receptor impaired mice: Model of depression [Abstract in Rivista]
Alboni, Silvia; Corsini, Daniela; Caggia, Federica; Benatti, Cristina; Capone, Giacomo; Barden, N; Blom, Johanna Maria Catharina; Tascedda, Fabio; Brunello, Nicoletta
abstract

Objective: The gene codifying for the neurotrophin Brain Derived Nurotrophic Factor (BDNF) is a stress-responsive gene and alteration in its expression may be important in producing some of the pathophysiological effects of stress in the hippocampus as seen in stress-related pathologies like depression. While the effects of stress procedures on the regulation of BDNF expression was widely investigated in hippocampus of healthy control animals, the stress-induced effects on BDNF hippocampal expression in “pathological” condition are still lacking. In order to deepen our knowledge in the understanding of the effects of an acute stressful procedure on molecular targets of synaptic plasticity we used transgenic mice with impaired glucocorticoid receptor (GR-i) expression that represent an animal model of depression. The hypothesis was tested that a single period of restraint stress (6 hours) affects BDNF mRNA expression in the hippocampus of GR-i mice differently than in wild-type (WT) mice. Methods: Using real time RT-PCR we evaluated the effects of a 6 hours acute stress on the levels of BDNF coding exon VIII and the activity regulated BDNF exon IV mRNA. In the WT and in the GR-i animals, the hippocampal levels of the two BDNF exons, immediately after the stress ended, were significantly lower in stressed animals with the respect to respective control unstressed animals. However, for the BDNF exon IV mRNA the reduction is most pronounced inWT animals and two-way ANOVA followed by Bonferroni posttest revealed a significant interaction between stress response and genotype at the level of BDNF exon IV mRNA expression. Results: The BDNF gene is a very complex gene regulated by a wide array of stimuli and signalling pathways. An electophoresis mobility shift assay (EMSA) was used to study DNA-binding activity of two transcription factors with an important role in controlling synaptic plasticity most likely trough an involvement BDNF: the cAMP responsive element binding (CREB) protein and the nuclear factor kB (Nf-kB). Taken together, our results show a different binding activity of these transcription factors in GR-i mice with respect to WT mice following acute stressful conditions. Conclusion: The identification of the molecular mechanisms activated by stress in GR-i mice model of depression may contribute to the development of new strategies that reducing neuron vulnerability to stress and prevent neurophatologocal alteration in the hippocampus.

2008 - Microarray analysis of the chronic escape deficit model of depression: Effects of escitalopram treatment in hippocampus [Abstract in Rivista]
Caggia, Federica; Valensisi, Cristina; Alboni, Silvia; Benatti, Cristina; Corsini, Daniela; Ferrari, F; Tagliafico, Enrico; Mendlewicz, J; Brunello, Nicoletta; Tascedda, Fabio
abstract

Objective: Currently, the biological bases of depression and the molecular mechanisms underlying antidepressant action are not completely understood. Behavioural models of depression and genome-wide gene expression analysis can be relevant to better understand the pathophysiology of this disease. Chronic escape deficit is a valid and useful model of depression and is based on the induction of an escape deficit after exposure of rats to unavoidable stress. This behavioural model allows to evaluate the capacity of a treatment to revert the escape deficit. The majority of antidepressant drugs need to be administered for at least 3−4 weeks in order to revert the escape deficit. In this study, we demonstrated that only one week of treatment with Escitalopram, a widely used SSRI, is effective in the chronic escape deficit model of depression. Also, our study demonstrated that only 50% of the animals receiving ESC responded to the treatment. The mechanisms underlying the action of escitalopram are still poorly understood and the molecular targets and pathways involved remain to be identified. In order to identify the biological target involved in the response to escitalopram, we performed a microarray experiment using the chronic escape deficit model of depression after a 7 day treatment with escitalopram. Methods: Gene expression patterns in the rat hippocampus were analyzed using Affymetrix GeneChip Rat Exon 1.0 ST evaluating both gene-level and exon-level expression profiling on the whole genome. Total RNA extracted from hippocampus of each treated animal was utilized to chipping a single array using the Affymetrix protocols. 20 single arrays were utilized for data analysis and divided into five replicates for each experimental group (naive, stress, escitalopram responders and not responders). With two parallel analyses (gene level and exon level) of raw data files carried out in Expression Console software using iterPLIER algorithms, we identified various transcripts that were differentially regulated in each pairwise comparison. In order to identify biological processes and signalling networks regulated by escitalopram response, we performed a functional analysis using Ingenuity web tool. Results: Functional annotation of selected genes reflected interesting different biological features between escitalopram responders and not responders. More specifically, the biological functions regard cellular growth and proliferation, gene expression and signal transduction. Conclusion: We believe that this pharmacogenomic approach will be helpful to understand the molecular mechanisms involved in the pathogenesis of depression as well as in the response to antidepressant drugs.

2008 - Molecular effects of subchronic andchronic treatment with escitalopram inthe rat central nervous system [Abstract in Rivista]
Benatti, Cristina; Alboni, Silvia; Capone, Giacomo; Corsini, Daniela; Caggia, Federica; Blom, Johanna Maria Catharina; Tascedda, Fabio; Brunello, Nicoletta
abstract

A clear understanding of the mechanisms behind depressionand its treatment is a critical issue for amelioratethe effectiveness of existing antidepressants. Acutely,antidepressant drugs increase synaptic concentrations ofmonoamines, but clinical efficacy requires several weeksof continuous treatment, proposing a key role for timedependentneural adaptations, perhaps induced by acutesynaptic actions, in their therapeutic efficacy.Escitalopram is the S(+) enantiomer of citalopram, oneof the most widely prescribed serotonin selective reuptakeinhibitor (SSRIs) antidepressants. In the chronic mildstress model of depression sucrose intake was alreadynormalized after one week of treatment.We evaluated the effects of a subchronic or a chronictreatment with escitalopram on expression levels of someof the main targets of antidepressant drugs such as theneurotrophin Brain Derived Neurotrophic Factor (BDNF),the transcription factors cAMP Response Element Binding(CREB) [1] Protein and Calcium Responsive Factor(CaRF).Sprague-Dawley rats were treated for 7 days (subchronic)or 21 days (chronic) with either escitalopram(10 mg/kg die i.p) or saline (1 mL/kg die); BDNF, CREBand CaRF mRNAs were evaluated using RNAse ProtectionAssay in hippocampus and prefrontal cortex.No difference was observed on BDNF, CREB andCaRF expression in the hippocampus of rats treatedsubchronically with escitalopram with respect to the grouptreated with saline. In contrast a significant inductionof BDNF mRNA was observed in prefrontal cortexof escitalopram-treated animals with respect to salinetreated ones. CaRF expression patterns were similar.Escitalopram for 7 days caused a significant induction ofCaRF mRNA with respect to the group treated with saline(p<0.05; Dunnett t-test), on the other hand CREB mRNAremained unaffected. Following a chronic treatment withEscitalopram, BDNF, CREB and CaRF mRNA levels weresignificantly decreased with respect to the group treatedwith saline in hippocampus (p<0.05; Dunnett t-test),while a 21 day treatment with escitalopram failed toproduce changes in gene expression in prefrontal cortex.These results showed that escitalopram is able todifferentially affect BDNF, CREB and CaRF expressionwith respect to treatment duration and that the observedeffects are area-specific.Consequently, to further investigate the possiblemolecular mechanisms underlying the observed effectson gene expression we evaluated by western blottingsome of the main signalling pathways regulating CREB aswell as BDNF expression, such as p38 MAPK (Mitogen-Activated Protein Kinase), CaMKII (Calcium CalmodulineKinase), ERK 1/2 (Extracellular Signal-Regulated Kinase)and CREB itself [1].Our study demonstrates that:1. A subchronic treatment with escitalopram inducesBDNF and CaRF expression in prefrontal cortexprobably through activation of p38 MAPK signallingpathway.2. A 21 day escitalopram treatment reduces hippocampalBDNF, CaRF, CREB gene expression and also CREBphosphorylated nuclear levels.Spatially distinct signalling pathways may be involvedin mediating the differential effect on gene expressionobserved following either a subchronic or a chronictreatment with escitalopram.

2007 - Combined effect of antidepressant and anti-inflammatory drugs in an animal model of depressio [Abstract in Rivista]
Brunello, Nicoletta; Alboni, Silvia; Benatti, Cristina; Corsini, Daniela; Capone, Giacomo; Tascedda, Fabio; J., Mendlewicz
abstract

The delay in the onset of action of antidepressants remains one of the unsolved issues in the treatment of depression despite the availability of safe and effective drugs. It has been known for years that depression may share identical symptoms as those in inflammatory reaction and that immune function and inflammation markers are altered in psychiatric patients. This suggests to use anti-inflammatory drugs as an adjunctive therapy for depression. We previously demonstrated that the combination of acetylsalicylic acid (ASA) with Fluoxetine (FLX) accelerated and potentiated the effect of the antidepressant in the chronic escape model of depression (Brunello et al., 2006). These results, together with preliminary clinical data in major depressed nonresponder patients, suggest that ASA might accelerate the onset of action of SSRIs (Mendlewicz et al., 2006). Preclinical and clinical studies have recently reported that Escitalopram (ESC), the active enantiomer of citalopram, shows a faster onset of action compared to other antidepressants. The aim of our study was to compare the effect of one week of combined treatment with ESC plus ASA vs. ESC alone in the chronic escape deficit model of depression. Significant response after one week of treatment was present in about 50% of the animals receiving ESC (10 mg/kg/day) alone and in about 75% of the rats receiving ESC plus ASA (45 mg/kg/day). These results suggest that the co-administration of ASA with ESC increased the response to treatment in reverting the behavioural despair induced by stress in rats, thus confirming the specific effect of combined therapy.

2007 - The transporters GlyT2 and VIAAT cooperate to determine the vesicular glycinergic phenotype [Articolo su rivista]
Aubrey, Kr; Rossi, Fm; Ruivo, R; Alboni, Silvia; Bellenchi, Gc; Le Goff, A; Gasnier, B; Supplisson, S.
abstract

The mechanisms that specify the vesicular phenotype of inhibitory interneurons in vertebrates are poorly understood because the two main inhibitory transmitters, glycine and GABA, share the same vesicular inhibitory amino acid transporter (VIAAT) and are both present in neurons during postnatal development. We have expressed VIAAT and the plasmalemmal transporters for glycine and GABA in a neuroendocrine cell line and measured the quantal release of glycine and GABA using a novel double-sniffer patch-clamp technique. We found that glycine is released from vesicles when VIAAT is coexpressed with either the neuronal transporter GlyT2 or the glial transporter GlyT1. However, GlyT2 was more effective than GlyT1, probably because GlyT2 is unable to operate in the reverse mode, which gives it an advantage in maintaining the high cytosolic glycine concentration required for efficient vesicular loading by VIAAT. The vesicular inhibitory phenotype was gradually altered from glycinergic to GABAergic through mixed events when GABA is introduced into the secretory cell and competes for uptake by VIAAT. Interestingly, the VIAAT ortholog from Caenorhabditis elegans (UNC-47), a species lacking glycine transmission, also supports glycine exocytosis in the presence of GlyT2, and a point mutation of UNC-47 that abolishes GABA transmission in the worm confers glycine specificity. Together, these results suggest that an increased cytosolic availability of glycine in VIAAT-containing terminals was crucial for the emergence of glycinergic transmission in vertebrates.

2006 - Acetylsalicylic acid accelerates the antidepressant effect of fluoxetine in a rat model of depression [Abstract in Rivista]
Capone, Giacomo; Alboni, Silvia; Benatti, Cristina; Tascedda, Fabio; Blom, Johanna Maria Catharina; J., Mendlewicz; Brunello, Nicoletta
abstract

Depression currently ranks fourth among the major causesof disability worldwide and by 2020, it is estimated thatunipolar major depression will rank second as a sourceof lost disability-adjusted life years (DALYs) worldwide(Murray and Lopez, 1997). To date however no singleagent is effective in all patients treated, probably due tothe different neurobiological alterations occurring for thedisorder and to individual differences in pharmacogeneticand pharmacodynamic parameters.Different therapies have been proposed to amelioratethe clinical responses of antidepressant drugs throughaugmentation or combination strategies.Another achievement in the development of newtreatments is to reduce the latency of clinical effect ofantidepressant drugs known to be characterized by 4-6weeks lag phase.Evidence has accumulated suggesting that majordepression is associated with dysfunction of inflammatorymediators and that psychiatric symptoms may occur duringinflammatory diseases. Moreover antidepressants show ananti-inflanmlatory action possibly related to their clinicalefficacy. In fact, an improvement in psychiatric symptomshas been recently reported in patients treated with antiinflammatorydrugs for other indications.These data imply that inflammation may be involved inthe pathogenesis of depression and that anti-inflanm~atorydrugs may be used as an adjunctive therapy.Among anti-inflammatory drugs acetylsalicylic acid(ASA) has been shown to act on the brain serotonergicsystem and to have a neuroprotective effect in vitro aswell as in vivo.Aim of the present study was to evaluate the effectsof combined treatment fluoxetine (FLX) plus ASA andASA alone in a behavioural model of depression:the chronic escape deficit (Gambarana et al., 2001).The chronic escape deficit model is based on themodified reactivity of rats to external stimuli inducedby exposure to unavoidable stress and allows evaluatingthe capacity of a treatment to revert the condition ofescape deficit. In this model, FLX alone needs to beadministered for at least 3 weeks in order to revert thiscondition.Our results showed that ASA (45 mg/kg) did not possessantidepressant properties in the chronic escape deficitmodel at any time tested. A combined treatment of FLX(5 mg/kg) and ASA (45 mg/kg) completely reverted thecondition of escape deficit as early as after 7 days, theeffect being already partially present after 4 days. Theeffect was maintained after 14 and 21 days of treatment.In the same experimental condition the effect of FLX(5 mg/kg) was significant only at 21 days, as previouslydemonstrated by other groups.The exact nature of the mechanisms underlying theabove behavioural effects are still unknown, neverthelessseveral hypotheses can be formulated. Further biochemicaland genetic researches could help to clarify the targets ofaction of the combined treatment FLX plus ASA for thedevelopment of more active and faster acting molecules.

2006 - Acetylsalicylic acid accelerates the antidepressant effect of fluoxetine in the chronic escape deficit model of depression [Articolo su rivista]
Brunello, Nicoletta; Alboni, Silvia; Capone, Giacomo; Benatti, Cristina; Blom, Johanna Maria Catharina; Tascedda, Fabio; Kriwin, P; Mendlewicz, J.
abstract

Evidence has accumulated suggesting that major depression is associated with dysfunction of inflammatory mediators. Moreover, antidepressants show an antiinflammatory action possibly related to their clinical efficacy. An improvement in psychiatric symptoms has been recently reported in patients treated with antiinflammatory drugs for other indications. These data imply that inflammation may be involved in the pathogenesis of depression and that anti-inflammatory drugs may be used as an adjunctive therapy. The aim of the present study was to evaluate the behavioural effect of the co-administration of acetylsalicylic acid (ASA, 45 mg/kg or 22.5 mg/kg) and fluoxetine (FLX, 5 mg/kg) in the chronic escape deficit model of depression. The chronic escape deficit model is based on the modified reactivity of rats to external stimuli induced by exposure to unavoidable stress and allows evaluation of the capacity of a treatment to revert the condition of escape deficit. In this model, FLX alone needs to be administered for at least 3 weeks to revert this condition. Our results show that combined treatment of fluoxetine and ASA completely reverted the condition of escape deficit by as early as 7 days, the effect being already partially present after 4 days. The effect was maintained after 14 and 21 days of treatment. ASA alone was ineffective at any time tested and the effect of fluoxetine was significant only at 21 days. These results, together with clinical data from preliminary results, suggest that ASA might accelerate the onset of action of selective serotonin reuptake inhibitor antidepressants.

2006 - Behavioural and molecular effects of the combined treatment fluoxetine plus acetylsalicylic acid in a rat model of depression [Abstract in Rivista]
Alboni, Silvia; Capone, Giacomo; Benatti, Cristina; Tascedda, Fabio; Blom, Johanna Maria Catharina; Mendlewicz, J; Brunello, Nicoletta
abstract

Current treatments for depression are inadequate for many patients, and different therapies have been proposed to ameliorate the clinical responses of antidepressant drugs through augumentation or combination strategies. Another achievement in the development of new treatments is to reduce the latency of clinical effect of antidepressant drugs known to be characterized by 4−6 weeks lag phase. Evidence has accumulated suggesting that major depression is associated with dysfunction of inflammatory mediators and that psychiatric symptoms may occur during inflammatory diseases. Moreover antidepressants show an anti-inflammatory action possibly related to their clinical efficacy. In fact, an improvement in psychiatric symptoms has been recently reported in patients treated with anti-inflammatory drugs for other indications. These data imply that inflammation may be involved in the pathogenesis of depression and that anti-inflammatory drugs may be used as an adjunctive therapy. Among the anti-inflammatory drugs the Acetylsalicylic acid has been shown to act on the brain serotonergic system and to have a neuroprotective effect toward brain damage. Aim of the present study was to evaluate the effects of combined treatment fluoxetine (FLX) plus acetylsalicylic acid (ASA) in a behavioural model of depression: the chronic escape deficit. The chronic escape deficit model is based on the modified reactivity of rats to external stimuli induced by exposure to unavoidable stress and allows evaluating the capacity of a treatment to revert the condition of escape deficit. In this model, FLX alone needs to be administered for at least 3 weeks in order to revert this condition. Our results showed that ASA (45 mg/kg) did not possess antidepressant properties in the chronic escape deficit model at any time tested. A combined treatment of FLX (5 mg/kg) and ASA (45 mg/kg) completely reverted the condition of escape deficit as early as after 7 days, the effect being already partially present after 4 days. The effect was maintained after 14 and 21 days of treatment. In the same experimental condition the effect of FLX (5 mg/kg) was significant only at 21 days, as previously demonstrated by other groups. The exact nature of the mechanisms underlying the above behavioural effects are still unknown, nevertheless several hypotheses can be formulated. Moreover, because a role for the neurotrophin BDNF was proposed in the clinical response to antidepressant treatment, we have determined the effect of combined treatment FLX plus ASA on hippocampal BDNF mRNA and protein in the same behavioural model. Ours data demonstrated that the hippocampal levels of BDNF mRNA were significantly increased with respect to control groups (naive and stressed) only in the animals responding (number of escape 10 out of 30 trials) to the combined treatment. Further biochemical and genetic researches could help to clarify the targets of action of the combined treatment FLX plus ASA for the development of more active and faster molecules.

2006 - Early postnatal chronic inflammation produces long-term changes in pain behavior and N-methyl-D-aspartate receptor subtype gene expression in the central nervous system of adult mice [Articolo su rivista]
Blom, Johanna Maria Catharina; Benatti, Cristina; Alboni, Silvia; Capone, Giacomo; Ferraguti, Chiara; Brunello, Nicoletta; Tascedda, Fabio
abstract

The objective of this study was to test whether postnatal chronic inflammation resulted in altered reactivity to pain later in life when reexposed to the same inflammatory agent and whether this alteration correlated with brain-region-specific patterns of N-methyl-D-aspartate (NMDA) receptor subtype gene expression. Neonatal mouse pups received a single injection of complete Freund's adjuvant (CFA) or saline into the left hind paw on postnatal day 1 or 14. At 12 weeks of age, both neonatal CFA- and saline-treated animals received a unilateral injection of CFA in the left hind paw. Adult behavioral responsiveness of the left paw to a radiant heat source was determined in mice treated neonatally with saline or CFA before and after receiving CFA as adults. Twenty-four hours later, brains were dissected and NMDA receptor subunit gene expression was determined in four different brain areas by using an RNase protection assay. The results indicated that NMDA receptor subtype gene expression in adult mice exposed to persistent neonatal peripheral inflammation was brain region specific and that NMDA gene expression and pain reactivity differed according to the day of neonatal CFA exposure. Similarly, adult behavioral responsiveness to a noxious radiant heat source differed according to the age of neonatal exposure to CFA. The data suggest a possible molecular basis for the hypothesis that chronic persistent inflammation experienced early during development may permanently alter the future behavior and the sensitivity to pain later in life, especially in response to subsequent or recurrent inflammatory events

2006 - Effects of acute stress on brain-derived neurotrophic factor in the hippocampus of transgenic mouse model of depression [Abstract in Rivista]
Alboni, Silvia; Blom, Johanna Maria Catharina; Corsini, Daniela; Benatti, Cristina; Capone, Giacomo; Ferraguti, Chiara; N., Barden; Tascedda, Fabio; Brunello, Nicoletta
abstract

Brain-Derived Neurotrophic Factor (BDNF) is a member of the neurotrophin family which includes a group of molecules important for the development and the maintenance of the nervous system. Since BDNF is highly expressed in the hippocampus, the action and regulation of this neurotrophin in this area has become subject of intense study. The gene codifying for BDNF is a stress responsive gene and alterations in its expression may be important in regulating some of the physiological and pathophysiological effects of chronic and acute stress in the hippocampus. Different studies show that several types of stress reduce BDNF expression in the hippocampus of control animals [Smith et al., 1995] and these works led to a neurotrophic hypothesis of depression [Nestler et al., 2002]. Nevertheless, the effect of stress on BDNF gene expression may differ between a "normal" and a "pathological" brain. In our study, we used transgenic mice with glucocorticoid receptor impaired (GRi) expression created [Pepin et al., 1992], as a tool to study the neuroendocrine changes observed in stress-related disorders, such as major depression. This GRi mouse model is characterized by dysfunctional glucocorticoid inhibitory feedback and an excessive activation of the hypothalamic pituitar~adrenal (HPA) axis, that can be restored by antidepressant drugs' treatment. The hypothesis was tested that a single period of 30 minutes of restraint stress affects BDNF expression in the hippocampus of GRi mice differently than in wildtype (WT) mice. Using RNase protection assay and in situ hybridization we had assessed the BDNF mRNA hippocampal levels, while the levels of BDNF and its precursor, pro-BDNF were analyzed by western blotting. Our results indicated that 30 minutes of restraint enhanced BDNF mRNA expression in the CA3 hippocampal subregion of GRi mice; the same stress procedure induced also a statistically significant increase of pro- BDNF level in hippocampus of GRi mice. No effect of acute stress was observed in the WT at the level of the expression of BDNE Moreover, we evaluated the effects of restraint on signalling pathways implicated in the regulation of BDNF expression (mitogen-activated protein kinase and calcium/calmodulin-dependent kinase cascades) that converge on the phosphorylation of CREB that we found down-regulated in the hippocampus of GRi mice and up-regulated in WT mice. Our data suggest that, in the presence of psychophysiological stress (restraint stress), GRi mice display altered hippocampal regulation in BDNF gene expression. Thus, life-long central GR dysfunction may negatively affects neural functioning by limiting the capacity to cope with change or acute stress, which could be a predisposing or determining factor in depression. Understanding the mechanisms underlying the induction of BDNF mRNA and accumulation of pro-BDNF in the hippocampus of GRi mice, may help to clarify the molecular basis of action of this neurotrophin and contribute to the development of new strategies reducing the vulnerability of neurons to stress, thus preventing neuropathological alterations in the hippocampus.

2006 - Shortened onset of action of antidepressants in major depression using acetylsalicylic acid augmentation: a pilot open-label study [Articolo su rivista]
Mendlewicz, J; Kriwin, P; Oswald, P; Souery, D; Alboni, Silvia; Brunello, Nicoletta
abstract

Based on our preclinical data showing a potential accelerating effect of acetylsalicylic acid (ASA) in combination with fluoxetine in an animal model of depression, we examined the effect of ASA augmentation therapy on selective reuptake inhibitors (SSRI) in major depressed non-responder patients. Twenty-four non-responder patients having received at least 4 weeks of an adequate SSRI treatment were included in a pilot open-label study. Participants were treated openly during 4 weeks with 160 mg/day ASA in addition to their current antidepressant treatment. The combination SSRI-ASA was associated with a response rate of 52.4%. Remission was achieved in 43% of the total sample and 82% of the responder sample. In the responder group, a significant improvement was observed within week 1 (mean Hamilton Depression Rating Scale-21 items at day 0 = 29.3 +/- 4.5, at day 7 = 14.0 +/- 4.1; P < 0.0001) and remained sustained until day 28. Despite limitations due to the open nature of this study, our preliminary results confirm our preclinical findings and are in favour of an accelerating effect of ASA in combination with SSRIs in the treatment of major depression. Potential physiological and biochemical mechanisms may involve an anti-inflammatory and/or neurotrophic effect.

2005 - Neonatal persistent inflammation alters pain response and NMDA receptor expression in adult mice [Abstract in Rivista]
Benatti, Cristina; Alboni, Silvia; Ferraguti, Chiara; Tascedda, Fabio; Blom, Johanna Maria Catharina; Brunello, Nicoletta
abstract

Infant pain is of critical interest, especially with respect to premature infants and other high-risk neonates that experience many invasive and traumatic procedures early in development. The early neonatal period is characterized by great plasticity and reorganization. Sustained activation of central nervous circuits, caused by protracted and recurrent pain, may cause long-lasting changes in central neural function thus affecting developmental outcome and behavioural responsiveness to pain or stress later in life. However little is known about the neurobiological substrates underlying this ``memory'' process. The aim of our study was twofold:to study whether timing of postnatal exposure to a persistent inflammatory insult alters the responsiveness to thermal pain in the adult animal;given the role of the NMDA receptor in pain processing as well as in learning and memory, to examine if NMDA receptor subtype gene expression in specific areas of the cns is influenced by neonatal inflammation.Methods: Newborn mice received a single injection of Complete Freund's Adjuvant (CFA) or saline on either postnatal day 1, 3 or 14 (P1, P3 and P14) into the left hind paw. At twelve weeks of age paw withdrawal latency (PWL) of each animal was tested both in basal condition and 24h after an unilateral injection of 100 μL of CFA in the left hind paw. Mice were then killed by cervical dislocation and cerebral areas were removed. Using a sensitive RNAse protection assay, NMDA receptor subunit (NR1, NR2A, NR2B, NR2C) gene expression was evaluated in different brain areas; all data were processed by one-way ANOVA (p < 0.05).Results: Baseline paw withdrawal latency was significantly decreased in animals exposed to CFA at day 1 and 14 as compared to their saline exposed counterparts. Animals exposed to CFA at postnatal day 3 showed a significant increase in paw withdrawal latency with respect to saline injected animals. Twenty-four hours later a unilateral injection of CFA into the left hind paw, a significant decrease in paw withdrawal latency was observed in all experimental groups with respect to baseline values. PWL of P1 saline treated animals after CFA exposure was significantly higher than P3 and P14 saline treated mice. Adult mice exposed to an injection with CFA on postnatal day 1 exhibited reduced expression of the NMDA receptor subtype NR1 and NR2C in the hippocampus while mRNA levels for NR2A and NR2B did not differ between CFA treated and untreated mice. Exposure to CFA on postnatal day 3 and 14 did not affect adult expression levels of NMDA receptor subunits in the hippocampus. NMDA receptor subunit expression displayed a different profile in the thalamus. Exposure to CFA at P1 and P3 did not alter NMDA receptor subunit expression while exposure to CFA at P14 resulted in enhanced expression of the NR2A and NR2B subunits.Conclusions: These findings indicate that changes in NMDA receptor subtype gene expression in adult mice exposed to persistent neonatal peripheral inflammation are brain region specific and that NMDA gene expression and pain reactivity differ according to the day of neonatal exposure to CFA.

2005 - New combination therapies from animal to human [Abstract in Rivista]
Brunello, Nicoletta; Alboni, Silvia; Benatti, Cristina; Capone, Giacomo; Tascedda, Fabio; Blom, Johanna Maria Catharina; J., Mendlewicz
abstract

Evidence has accumulated suggesting that major depression is associated with dysfunction of inflammatory mediators. Antidepressants interfere with the synthesis and release of cytokines and do not exert behavioral effects in animal models of depression when hippocampal neurogenesis is blocked, a phenomenon which is occurring in the presence of inflammation. The anti-inflammatory drug acetylsalicylic acid (ASA), besides inhibiting the cyclooxigenase pathway, interacts with central serotonergic system, by increasing serotonin levels in cortex and reducing the density of different serotonin receptor subtypes. These neurochemical effects suggest a role of ASA in the treatment of depression. Therefore we studied the effect of ASA and fluoxetine combined treatment in a behavioral model of depression. The chronic escape deficit model is based on the modified reactivity of rats to external stimuli induced by exposure to unavoidable stress and allows evaluating the capacity of a treatment to revert the condition of escape deficit. Any kind of antidepressant drug needs to be administered for at least 3 weeks in order to revert this condition. The combined treatment of fluoxetine and ASA completely reverted the condition of escape deficit as early as after 7 days, the effect being already partially present after 4 days. The effect was maintained after 14 and 21 days of treatment. ASA alone was ineffective at any time tested and the effect of fluoxetine was significant only at 21 days. Given these preclinical results, an open clinical study has been started using the combination SSRI-ASA in treatment resistant depressed patients. Preliminary results suggest a potential accelerating effect of ASA in combination to SSRI.

2004 - Regulation of CREB function in rat frontal cortex after combined treatment with Fluoxetine and Olanzapine [Abstract in Rivista]
Capone, Giacomo; Alboni, Silvia; Blom, Johanna Maria Catharina; Ferraguti, Chiara; Brunello, Nicoletta; Tascedda, Fabio
abstract

Statement of the Study: Generally, drugs used in the treatment of depression exert their therapeutic effect after 4/6 weeks and only in 60–65% of patients. The search for an adequate and faster treatment of major depression is one of the main challenges in neuropsychopharmacology. Recently, a clinical study of treatmentresistant depressed patients without a psychotic component, showed that after only one week of treatment, Fluoxetine (a selective serotonin reuptake inhibitor antidepressant) and Olanzapine (an atypical antipsychotic agent) produced a higher level of improvement than either monotherapy alone (Shelton et al., American Journal of Psychiatry 158(1), 131–134, 2001). Furthermore, preclinical data, using microdialysis, indicated that the combination of Olanzapine and Fluoxetine resulted in an increase in the extracellular levels of dopamine and norepinephrine in the rat prefrontal cortex, an effect that was significantly bigger than after treatment with either drug alone (Zhang et al., Neuropsychopharmacology 23(3),250– 262, 2000). However, it is not yet completely understood which intracellular signaling pathway could be involved in the fast response (seen in clinical trials) to Fluoxetine plus Olanzapine co-treatment. Methods: Since antidepressant and antipsychotic drugs affect the cyclic adenosine monophosphate (cAMP) pathway, including the expression of the cAMP response element binding protein (CREB), the levels of CREB mRNA and CREB nuclear protein, total and phosphorylated, were studied in the frontal cortex of rats using RNase protection assay and Western Blotting analysis respectively. Four experimental groups were used: rats were treated for one, five or ten days with either saline, Fluoxetine (ip 10 mg/Kg), Olanzapine (sc 1 mg/Kg) or combined Fluoxetine plus Olanzapine (10 mg/Kg and 1 mg/Kg). Summary of Results: Our results show that the level of phosphorylated CREB Ser133 was significantly increased in the frontal cortex of rats receiving the combined treatment regimen (Fluoxetine plus Olanzapine) for five days. No effect was observed in acutely and ten day treated rats. Conclusion: This specific effect on CREB phosphorylation levels after five days of combined treatment with Fluoxetine plus Olanzapine might represent one of the mechanisms underlying the faster response to this therapy recently observed in several clinical trials.

2004 - Restraint stress increases the expression of brain derived neurotrophic factor in the hippocampus of a mouse model of depression [Abstract in Rivista]
Alboni, Silvia; Benatti, Cristina; Blom, Johanna Maria Catharina; Ferraguti, Chiara; Tascedda, Fabio; Barden, N; Brunello, Nicoletta
abstract

Statement of the study: Brain-Derived Neurotrophic Factor (BDNF) is a member of the neurotrophin family which includes a group of molecules important for the development and the maintenance of the nervous system. Since BDNF is highly expressed in the hippocampus, the action and regulation of BDNF in this particular area has become subject of intense study. Single or repeated immobilization stress markedly reduces both BDNF mRNA and protein levels in rat hippocampus. Consequently, BDNF is considered a stress-responsive gene, and it has been recently suggested that alterations in the expression of this growth factor may be important in regulating some of the physiological and pathophysiological effects of stress on the hippocampus. Stress-induced changes observed in the hippocampus of experimental animals resulted in a novel hypothesis attributing a central role to neurotrophic factors in both the etiology of depression and as well as in its treatment. However, the effects of stress on neurotrophic factors in the hippocampus of depressed patients remain unknown. In fact, the expression pattern of a large array of genes affected by depression or antidepressant drugs, such as BDNF, may differ between a normal and a pathological brain. Methods: In these experiments, we used transgenic (TG) mice deficient in glucocorticoid receptor (GR) functioning. This TG mouse was created as a model to study the neuroendocrine changes occurring in stress-related disorders, such as major depression. We evaluated the hypothesis that a single period of 30 minutes of restraint stress affects BDNF mRNA expression in the hippocampus of TG mice differently than in WT mice. Summary of results: BDNF mRNA was significantly increased by the stress procedure only in the hippocampus of TG mice, the induction being specific for the CA3 subregion as revealed by in situ hybridization. Moreover, we found that stress down-regulated CREB phosphorylation in the hippocampus of TG mice whereas it upregulated the level of phosphorylated CREB Ser133 in WT mice. Conclusion: These data suggest that, in the presence of emotional stress, lifelong central glucocorticoid receptor dysfunction results in altered hippocampal sensitivity, with respect to the level of neurotrophic gene expression. The understanding of the mechanisms through which psychological stress (such as restraint stress) induces BDNF mRNA in the hippocampus of TG mice, may help to clarify the biological and molecular basis of the action of neurotrophic factors and may contribute to the development of new strategies that will ultimately reduce the vulnerability of neurons and prevent neuropathological alterations in the hippocampus.