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FEDERICA CAGGIA
DIPENDENTE AZIENDA POLICLINICO
Dipartimento di Scienze Mediche e Chirurgiche Materno-Infantili e dell'Adulto
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Pubblicazioni
2023
- Targeting PI3K/AKT/mTOR Pathway in Breast Cancer: From Biology to Clinical Challenges
[Articolo su rivista]
Cerma, K.; Piacentini, F.; Moscetti, L.; Barbolini, M.; Canino, F.; Tornincasa, A.; Caggia, F.; Cerri, S.; Molinaro, A.; Dominici, M.; Omarini, C.
abstract
Breast cancer (BC) is the most common women cancer and cause of cancer death. Despite decades of scientific progress in BC treatments, the clinical benefit of new drugs is modest in several cases. The phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway mutations are frequent in BC (20-40%) and are significant causes of aggressive tumor behavior, as well as treatment resistance. Improving knowledge of the PI3K/AKT/mTOR pathway is an urgent need. This review aims to highlight the central role of PI3K-mTORC1/C2 mutations in the different BC subtypes, in terms of clinical outcomes and treatment efficacy. The broad base of knowledge in tumor biology is a key point for personalized BC therapy in the precision medicine era.
2022
- Predictive factors for relapse in triple-negative breast cancer patients without pathological complete response after neoadjuvant chemotherapy
[Articolo su rivista]
Toss, A.; Venturelli, M.; Civallero, M.; Piombino, C.; Domati, F.; Ficarra, G.; Combi, F.; Cabitza, E.; Caggia, F.; Barbieri, E.; Barbolini, M.; Moscetti, L.; Omarini, C.; Piacentini, F.; Tazzioli, G.; Dominici, M.; Cortesi, L.
abstract
IntroductionTriple-negative breast cancer (TNBC) patients who do not obtain pathological complete response (pCR) after neoadjuvant chemotherapy (NACT) present higher rate of relapse and worse overall survival. Risk factors for relapse in this subset of patients are poorly characterized. This study aimed to identify the predictive factors for relapse in TNBC patients without pCR after NACT. MethodsWomen with TNBC treated with NACT from January 2008 to May 2020 at the Modena Cancer Center were included in the analysis. In patients without pCR, univariate and multivariable Cox analyses were used to determine factors predictive of relapse. ResultsWe identified 142 patients with a median follow-up of 55 months. After NACT, 62 patients obtained pCR (43.9%). Young age at diagnosis (<50 years) and high Ki-67 (20%) were signi!cantly associated with pCR. Lack of pCR after NACT resulted in worse 5-year event-free survival (EFS) and overall survival (OS). Factors independently predicting EFS in patients without pCR were the presence of multifocal disease [hazard ratio (HR), 3.77; 95% CI, 1.45-9.61; p=0.005] and residual cancer burden (RCB) III (HR, 3.04; 95% CI, 1.09-9.9; p=0.04). Neither germline BRCA status nor HER2-low expression were associated with relapse. DiscussionThese data can be used to stratify patients and potentially guide treatment decision-making, identifying appropriate candidates for treatment intensi!cation especially in neo-/adjuvant setting.
2022
- T-DM1 efficacy in trastuzumab-pertuzumab pre-treated HER2 positive metastatic breast cancer patients: a meta-analysis.
[Articolo su rivista]
Omarini, C; Piacentini, F; Sperduti, I; Cerma, K; Barbolini, M; Canino, F; Nasso, C; Isca, C; Caggia, F; Dominici, M; Moscetti, L.
abstract
Background: Current guidelines consider T-DM1 the standard 2nd line therapy for HER2 positive metastatic breast cancer (MBC) patients following trastuzumab (T) + pertuzumab (P) and taxane 1st line treatment. Despite this, there are no prospective studies supporting this sequence.
Methods: We performed a meta-analysis using real world data to determine the efficacy of T-DM1 after 1st line TP in HER2 positive MBC patients. We used a random-effect model to find differences in the rate of 1-year progression free survival (PFS) between TP pre-treated population and the EMILIA phase III pivotal trial.
Results: Seven studies were eligible. The meta-analysis showed a combined 1-year PFS risk difference for T-DM1 efficacy after TP in 2nd or more lines of -0.122, with lower and upper limits of -0.253 and 0.010, respectively (p = 0.07), with low heterogeneity among studies (I2 0.01%, p = 0.836). Considering the four studies on T-DM1 in 2nd line setting, 1-year PFS risk was -0.034 (95% CI -0.207 - 0,139; p = 0.701) (I2 0.01%, p = 0.91).
Conclusion: Overall, the efficacy of T-DM1 after TP seems to be similar to that previously reported in the EMILIA trial. In the second line setting, data are not mature enough to confirm T-DM1 efficacy in TP pre-treated population.
2021
- Adjuvant exemestane or
tamoxifen plus ovarian
suppression in premenopausal
women: single institution
analysis
[Abstract in Rivista]
D'Onofrio, R.; Piacentini, F.; Barbolini, M.; Isca, C.; Nasso, C.; Caggia, F.; Dominici, M.; Moscetti, L.; Omarini, C.
abstract
Background: The combined analysis of data from TEXT
and SOFT trials shows that among premenopausal women
with hormone receptor-positive (HR+) breast cancer
(BC), adjuvant endocrine therapy (AET) with exemestane
(EXE) plus ovarian function suppression (OFS) improved
disease-free survival compared to tamoxifen (TAM) plus
OFS. We conducted a single institution analysis to compare
the activity and safety of both treatment strategies.
Patients And Methods: The data on tumor and patient’s
characteristics of premenopausal women treated with AET
from January 2014 to December 2018 in our institution
were retrospectively collected. Treatment toxicities were
graded according to CTCAE v5. Survival data were analyzed
by Kaplan Meier curves and log rank test.
Results: 237 patients were included in the study: 120 on
TAM / OFS and 117 on EXE/OFS. Notably, 43 patients
(18%) started AET in 2014 (before TEXT/SOFT data):
93% of these were treated with TAM/OFS versus only 7%
with EXE/OFS. Women on EXE/OFS had more high-risk
early BC compared to those on TAM/OFS (STAGE III
23,9% vs 6,6%; luminal B-like 34,2% vs 21,6%; T> 2 cm
68,4% vs 32,5%; nodal status positive 66,6% vs 36,6% -
all p value <0,01). According with risk of relapse, the
number of patients pre-treated with chemotherapy was
higher in EXE/OFS group (79,5% versus 37,5%, p value
<0,001) than TAM/OFS one. Extended therapy was
accepted by 50% of patients in the TAM/OFS group and
47% in the EXE/OFS group. Any grade adverse events
(AE) were observed in 77 (64%) and 101 (86%) patients in
TAM/OFS and EXE/OFS group, respectively. In particular,
the incidence of G3 AEs was significantly higher in the
EXE/OFS group and mainly represented by muscoloskeletal
symptoms, osteoporosis and hypertension. Eighteen
(15,4%) women discontinued EXE and switched to an
alternate ET (TAM or NSAI) due to treatment toxicity. No
statistically significant difference in terms of relapse freesurvival
was observed between the two groups.
A – Breast Cancer 27
Conclusions: In our analysis, the choice of the AET is
driven mainly from the risk of relapse. EXE/OFS represents
the main choice in the high-risk patients as per SOFT/
TEXT trials results. TAM/OFS represent the main chose
antecedent to the SOFT/TEXT results (2014/2015).The
frequency and the grade of AEs were higher in EXE group
than TAM one. The AET should be proposed based on
both, risk of relapse and treatment toxicity profile. An
update analysis will be presented at the meeting.
2021
- Clinicopathologic Profile of Breast Cancer in Germline ATM and CHEK2 Mutation Carriers
[Articolo su rivista]
Toss, Angela; Tenedini, Elena; Piombino, Claudia; Venturelli, Marta; Marchi, Isabella; Gasparini, Elisa; Barbieri, Elena; Razzaboni, Elisabetta; Domati, Federica; Caggia, Federica; Grandi, Giovanni; Combi, Francesca; Tazzioli, Giovanni; Dominici, Massimo; Tagliafico, Enrico; Cortesi, Laura
abstract
The most common breast cancer (BC) susceptibility genes beyond BRCA1/2 are ATM and CHEK2. For the purpose of exploring the clinicopathologic characteristics of BC developed by ATM or CHEK2 mutation carriers, we reviewed the archive of our Family Cancer Clinic. Since 2018, 1185 multi-gene panel tests have been performed. Nineteen ATM and 17 CHEK2 mutation carriers affected by 46 different BCs were identified. A high rate of bilateral tumors was observed in ATM (26.3%) and CHEK2 mutation carriers (41.2%). While 64.3% of CHEK2 tumors were luminal A-like, 56.2% of ATM tumors were luminal B-like/HER2-negative. Moreover, 21.4% of CHEK2-related invasive tumors showed a lobular histotype. About a quarter of all ATM-related BCs and a third of CHEK2 BCs were in situ carcinomas and more than half of ATM and CHEK2-related BCs were diagnosed at stage I-II. Finally, 63.2% of ATM mutation carriers and 64.7% of CHEK2 mutation carriers presented a positive BC family history. The biological and clinical characteristics of ATM and CHEK2-related tumors may help improve diagnosis, prognostication and targeted therapeutic approaches. Contralateral mastectomy should be considered and discussed with ATM and CHEK2 mutation carriers at the first diagnosis of BC.
2021
- Next Generation Sequencing
(NGS): a possible game changer in
metastatic breast cancer
[Abstract in Rivista]
Barbolini, M.; Omarini, C.; Moscetti, L.; Canino, F.; Trudu, L.; Tornincasa, A.; Caggia, F.; Bettelli, S.; Manfredini, S.; Isca, C.; Molinaro, A.; Dominici, M.; Piacentini, F.
abstract
Background: NGS has been introduced into the clinics
with the aim of sequencing long and complex genes for
tumor sample, in order to identify driver and/or targetable
alterations. Several companies and academic centers have
implemented NGS assays to guide treatment decisions,
even though there are no clear recommendations from scientific
societies about their use in daily clinical practice.
Patients and methods: Since 2019 NGS analysis was performed
in 32 MBC patients’ tissues at Modena Cancer
Center, as for clinical judgement during the course of
MBC. Oncomine™ was mainly used for the assay. The
aim was to define the PI3K mutational status, since
Alpelisib - an a-subunit selective PI3K inhibitor - had
shown to improve PFS in PI3K mutated HR+/HER2-
MBC patients in SOLAR-1 and BYLIEVE trials.
Results: Twenty (62%) NGS analysis were performed on
MBC samples, the other (13) on primary breast cancer.
Table1 summarize the clinical-pathological characteristics
of patients. At least 1 mutation was found in 25 (78%)
samples. A PI3K mutation was detected in 14 (44%) cases,
with E542K as the most frequent. In 10 out of 14 cases,
PI3K mutation was associated with other gene mutations.
FGFR3, FGFR4 mutations and FGFR2 amplification were
described in 4, 2 and one patients respectively. Two
patients showed AKT1 mutation, in one case was associated
with PTEN mutation. Two of the patients with PI3K
mutation were treated with Alpelisib + Fulvestrant. The
patient with FGFR1 amplification was eligible for a phase
II clinical trial.
Conclusions: NGS performed in this cohort of MBC
patients allowed therapeutic decisions in about 10% of
cases. Although PI3K mutational status for eligibility to
Alpelisib can be cheaply studied by RT-PCR, NGS assay
can provide wider information about other gene mutations,
useful for patients’ selection for clinical trials. In the
era of precision medicine, knowing the mutational status
of MBC early in patient history can change the therapeutic
algorithm.
2021
- TDM-1 efficacy in trastuzumabpertuzumab
pre-treated HER2
positive metastatic breast cancer
patients: a meta-analysis
[Abstract in Rivista]
Omarini, C.; Piacentini, F.; Sperduti, I.; Cerma, K.; Barbolini, M.; Canino, F.; Nasso, C.; Isca, C.; Caggia, F.; Dominici, M.; Moscetti, L.
abstract
Background: Based on the results reported in Emilia trial
population, current guidelines consider TDM-1 the standard
2nd line therapy for HER2 positive metastatic breast
cancer (MBC) patients. Despite that, there are no prospective
studies supporting the efficacy of TDM-1 following
trastuzumab (T) + pertuzumab (P) and taxane 1st line
treatment. Currently, only real-world data have investigated
this sequence with controversial results.
Methods: We performed a meta-analysis of the available
real world data to determine the efficacy of T-DM1 after 1st
line TP in HER2 positive MBC patients. We used a random-
effect model to find differences in the rate of 1-year
progression free survival (PFS) between TP pre-treated
population and the phase III Emilia trial (T pre-treated
population).
Results: Seven studies were eligible, in three of them data
were from sub-group population analysis. The meta-analysis
showed a combined 1-years PFS risk difference for
TDM-1 efficacy after TP in 2nd or more lines of -0.122, with
lower and upper limits of -0.253 and 0.010, respectively
(p=0.07), with low heterogeneity among studies (I2 <
0.0001, p =0.836). Considering the four studies on TDM-1
in 2nd line setting, 1-years PFS risk was -0.034 (95% CI
-0.207 – 0,139; p=0.701) (I2 < 0.0001, p =0.91).
Conclusions: Results from the meta-analysis show that
the efficacy of TDM-1 after TP double-block seems to be
similar to the previously reported in Emilia trial. In the
second line setting, available data are not mature enough
to confirm TDM-1 efficacy in TP pre-treated population.
Currently, TP pretreated patients should receive T-DM1 as
indicated in the guidelines.
2019
- Clinical-Pathological Characteristics of HER2+ Breast Cancers patients among BRCA1/2+ carriers tested in Modena Cancer Center.
[Poster]
Barbolini, M; Omarini, C; Viola, L; Isca, C; Marchi, I; Caggia, F; Barbieri, E; Toss, A; Cortesi, L; Dominici, M; Piacentini, F.
abstract
Background and Objectives
Prevalence of HER2 positive breast cancer (BC) in BRCA1/2 carriers is still underestimated. Clinical behaviour of HER2+/BRCA+ BC could be in some ways different from sporadic HER2+ disease. The aim of this research is to describe clinical-pathological characteristics of this rare entity in patients treated at Modena Cancer Center.
Methods
Between January 2005 and July 2019, 2911 BC patients have been tested for BRCA1/2. 231 (7.9%) and 173 (5.9%) were found positive for BRCA1 and BRCA2 pathogenetic mutations respectively. We considered the HER2+ subgroup (14 patients), focusing on hormone receptor status, IGF-1R expression (semi-quantitative expression by immunohistochemistry with Anti-IGF-1R rabbit monoclonal), PI3K mutation (NGS Sequenom analysis) and clinical characteristics.
Results
All but one patients received trastuzumab as part of their treatment. Six patients were treated with neoadjuvant chemotherapy (NACT), comprehensive of trastuzumab +/- pertuzumab, achieving a pathological complete response in 33% of cases.
Two patients experienced disease recurrence after NACT (median EFS = 5.7 years).
From a pathological point of view, the majority of cancers were ductal infiltrating, grading 3 and hormone receptor positive.
3 cases a PI3K mutation was found: in one of them a double mutation (p.R38H + p.E545K) was detected.
IGF-1R showed high expression in 2 cases (IHC score 3+), intermediate expression in 2 cases (score 2+), low or absent expression in 3 cases.
Interestingly one HER2+/BRCA1 mutated patient shows both PI3K mutation and IGF-1R overexpression, achieving a partial response after NACT and still NED.
Conclusions
Here we report a low frequency of HER2+ BC in BRCA1/2 germline carriers. This rate is higher in patients with BRCA2 mutation, accordingly with previous literature. It is not clear, in this peculiar population that displays different possible pathogenetic drivers, which could be the main druggable
2018
- Clinical and molecular predictors of long-term response in HER2 positive metastatic breast cancer patients.
[Articolo su rivista]
Omarini, C; Bettelli, S; Caprera, C; Manfredini, S; Caggia, F; Guaitoli, G; Moscetti, L; Toss, A; Cortesi, L; Kaleci, S; Maiorana, A; Cascinu, S; Conte, Pf; Piacentini, F
abstract
BACKGROUND:
HER2+ metastatic breast cancer (MBC) is a poor prognosis disease, unusually curable. To date, no predictive factors have been clearly correlated with long-term response to anti-HER2 agents.
METHODS:
54 HER2+ MBC patients treated with HER2 targeted therapy as first line treatment were analysed: 40 with a time to progression longer than 3 years in Long Responders (LR) group and 14 with a progression disease within one year of anti-HER2 therapy in a control group named Early Progressors (EP). The expression of 770 genes and 13 molecular pathways were evaluated using Nanostring PanCancer pathway panel performed on FFPE BC tissues.
RESULTS:
Considering baseline patients and tumor characteristics, EP women had more CNS spread and more metastatic burden of disease compared to LR (p > 0.05). Gene expression analysis identified 30 genes with significantly different expression in the two cohorts; five were driver genes (BRCA1, PDGFRA, AR, PHF6 and MSH2). The majority of these genes were over-expressed, mainly in LR patients, and encoded growth factors, pro- or anti-inflammatory interleukins and DNA repair factors. Only four genes were down regulated, all in EP group (TNFSF10, CACNG1, IL20RB and BRCA1). Most of these genes were involved in MAPK and PI3K pathways. MAPK pathway was differently expressed between LR and EP (p = 0.05). PI3K was the only pathway overexpressed in EP patients.
CONCLUSIONS:
Whole genome expression analysis comparing LR vs. EP identified a group of genes that may predict more favourable long-term outcomes. Up-regulation of MAPK and down-regulation of PI3K pathway could be a positive predictive factors. Further clinical implications are warranted.
ABBREVIATIONS:
BC: breast cancer; MBC: metastatic breast cancer; LR: long responder; EP: early progressor; FFPE: formalin-fixed paraffin-embedded; CNS: central nervous system; PFS: progression free survival; OS: overall survival.
2018
- Impact of anaemia on tumor response to neoadjuvant chemotherapy in breast cancer patients .
[Poster]
Omarini, Claudia; Guaitoli, Giorgia; Barbolini, Monica; Moscetti, Luca; Balduzzi, Sara; Caggia, Federica; Cascinu, Stefano; Piacentini, Federico
abstract
BACKGROUND - Pathological complete response (pCR) to neoadjuvant systemic therapy (NST) in patients with breast cancer (BC) predicts long-term outcomes.
Anaemia is one of the most common side effects of cytotoxic drugs. Biologically, anaemia induces adaptive responses due to the low intra-tumoral oxygen levels that may be responsible for increase chemotherapy resistance. In literature, data regarding this issue are lacking.
AIM - To evaluate the influence of anaemia throughout treatment course on tumour shrinkage induced by NST. METHODS - Patients - 317 patients diagnosed with stage I-III BC treated with NST and with available blood tests were included. Patients and tumor characteristics and treatments information were collected. We focused on Haemoglobin (Hb) level (at baseline, at the end of NST, drop in Hb throughout treatment and duration of anaemia) and its correlation with pCR rate. Anaemia was defined as a drop of Hb under the local limit of normal in women (12 mg/dl).
Statistical analysis - Categorical variables were analyzed using chi-square test or Fisher's exact test, continuous variables using t test. Univariate and multivariate analyses were fit to determinate the association between anaemia and pCR rate. A p-value < 0.05 was considered statistically significant; hazard ratio was estimated with 95% of confidence limits. RESULTS- No difference in Hb levels was observed stratifying patients according to nuclear grade, tumor stage, cancer subtypes and chemotherapy regimens. Median baseline Hb was 13.3 g/dl while median Hb level at the end of NST was 10 g/dl. 31 patients had pre-treatment anaemia. 60% of patients developed anaemia during NST period. In the subgroup of anaemic patients, who had a decrease in Hb ≥ 2 g/dl from baseline or anaemia longer than two months, a lower rate of pCR was observed (16% vs 29%, p=0.03 and 16% vs 25%, p=0.01, respectively). Patients with both these characteristics had the lowest rate of pCR (10%, p=0.01). CONCLUSIONS - Anaemia is a negative predictive factor for tumor response in women treated with NST for BC. This evidence suggests that anaemia should be improved in order to improve response to NST.
2017
- Differential gene expression patterns in HER2 positive metastatic breast cancer patients according to hormone receptor status
[Poster]
Omarini, Claudia; Kaleci, Shaniko; Guaitoli, Giorgia; Bettelli, Stefania Raffaella; Cecilia, Caprera; Manfredini, Samantha; Caggia, Federica; Baschieri, MARIA CRISTINA; Luca, Moscetti; Maiorana, Antonino; Cascinu, Stefano; Piacentini, Federico
abstract
Background: HER2 positive breast cancer (HER2+ BC) is a heterogeneous disease. Presenting features, patterns of recurrence and survival of HER2+ BC can differ according to hormone receptors (HR) status. The purpose of this study is to highlight different gene profiles and molecular pathways between HR+ and HR- metastatic HER2+ BCs. Material and Methods: 34 HER2+ metastatic BC patients were included: 18 patients with HR+/HER2+ and 14 with HR-/HER2+. Data regarding tumor characteristics, treatment information and clinical outcomes were collected. The expression of 770 genes and 13 molecular pathways were evaluated by means of Nanostring PanCancer pathway panel performed on BC formalin-fixed paraffin-embedded tissues from diagnostic core biopsy or surgical resection. Results: Gene expression analysis identified 118 genes with significantly different expression in the two cohorts. All but one of these genes were over-expressed; only the gene CACNG6 was down-regulated in HR+/HER2+ group. In particular, 93 genes were over-expressed in HR-/HER2+ while 24 were overexpressed in HR+/HER2+. Most of these genes encoded growth factors, pro- or anti-inflammatory interleukins and DNA repair factors. 62% of these genes were involved in PI3K, MAPK and RAS pathways (32, 22 and 18, respectively). PI3K, MAPK and NOTCH pathways were differently expressed between HR+/HER2+ and HR-/HER2+ (p=0.003, p=0.0018, p=0.02, respectively). All these three pathways were overexpressed in HR-/HER2+ BC. In particular, all the significantly different expression genes in NOTCH pathways were overexpressed in HR-/HER2+ group. Conclusions: This genome expression analysis identified a gene expression profile able to differentiate HR+ versus HR- HER2+ metastatic BC. The overexpression of PI3K, MAPK and NOTCH pathways in HR-/HER2+ BC could justify its more aggressive behaviour. The validation of this HER2+ BC profile needs further investigation. Keywords: metastatic breast cancer, gene expression profile, HER2 positive breast cancer, PanCancer
2017
- Differential gene expression patterns in HER2 positive metastatic breast cancer patients according to hormone receptor status.
[Abstract in Rivista]
Omarini, C; Kaleci, S; Guaitoli, G; Bettelli, S; Caprera, Cecilia; Manfredini, S; Caggia, F; Baschieri, Cristina; Moscetti, Luca; Maiorana, A; Cascinu, S; Piacentini, F
abstract
Background: HER2 positive breast cancer (HER2+ BC) is a heterogeneous disease. Presenting features, patterns of recurrence and survival of HER2+ BC can differ according to hormone receptors (HR) status. The purpose of this study is to highlight different gene profile and molecular pathways between HR+ and HR- metastatic HER2+ BCs.
Materials and methods: 34 HER2+ metastatic BC patients were included: 18 patients with HR+/HER2+ and 14 with HR-/HER2+. Data regarding tumor characteristics, treatment information and clinical outcomes were collected. The expression of 770 genes and 13 molecular pathways were evaluated by means of Nanostring PanCancer pathway panel performed on BC formalin-fixed paraffin-embedded tissues from diagnostic core biopsy or surgical resection specimen.
Results: Gene expression analysis identified 118 genes with significantly different expression in the two cohorts. All but one of these genes were over-expressed; only the gene CACNG6 was down-regulated in HR+/HER2+ group. In particular, 93 genes were over-expressed in HR-/HER2+ while 24 were overexpressed in HR+/HER2+. Most of these genes encoded growth factors, pro- or anti-inflammatory interleukins and DNA repair factors. 62% of these genes were involved in PI3K, MAPK and RAS pathways (32, 22 and 18, respectively). PI3K, MAPK and NOTCH pathways were differently expressed between HR+/HER2+ and HR-/HER2 + (p = 0.003, p = 0.0018, p = 0.02, respectively). All these three pathways were overexpressed in HR-/HER2+ BC. In particular, all the significantly different expression genes in NOTCH pathways were overexpressed in HR-/HER2+ group.
Conclusions: This genome expression analysis identified a gene expression profile able to differentiate HR+ versus HR- HER2+ metastatic BC. The overexpression of PI3K, MAPK and NOTCH pathways in HR-/HER2+ BC could justify its more aggressive behaviour. The validation of this HER2+ BC profile needs further investigation.
2017
- Quality of life and anxious-depressive symptoms in cancer patients undergoing mindfulness-based interventions: Feasibility and preliminary outcomes on prospective single-centre case-control study (MIND4ME St.)
[Abstract in Rivista]
Pontoni, G.; Maur, Michela; Ferrari, Rita; Guida, Antonella; Poletti, S.; Caggia, Federica; Fontana, Annalisa; Gavioli, Francesco; Tarantino, Vittoria; Cascinu, Stefano; Ferrari, Silvia
abstract
Background
Mindfulness based interventions (MBIs) have shown efficacy in improving psychological symptoms including depression and anxiety in cancer patients (pts). The study aimed to explore feasibility and reproducibility of MBIs in an Italian Cancer Centre measuring biochemical and psychological parameters.
Methods
In this pilot prospective case-control study, we recruited newly diagnosed pts receiving adjuvant chemotherapy (CT). A MBIs program was designed consisting of 2.5 hours weekly for 8 weeks and, including meditation, yoga and body scan. Material for 45 minutes (mn) home daily practice was provided. Primary endpoint was to evaluate feasibility. Secondary endpoints were assessment of quality of life (QoL), psychological and biochemical outcomes of stress, tested at baseline (W0), W4, W8, W24, W48. PSS (Perceived Stress Reduction), POMS (profile of mood states scores), EuroQoL (EQ-5D-3L) were administered.
Results
Ten pts underwent MBIs program arm. We present preliminary results, while data of control arm are being collected. All pts were female, two pts (20%) dropped out. Median age was 56 years. All received adjuvant CT, 5/8 received radiotherapy and hormone therapy. Mean of sessions attending was 6.8 (76%). Median daily practice was 30 mn. EQ-5D item for depression and anxiety showed decreasing trend in mean score from moderate to light (P = 0.15) and significant improvement of auto-perceived QoL was observed comparing W0 and W8 (P = 0.02)
Conclusions
In a sensitive setting such as start CT, we found high pts compliance to MBIs. Improvement in self-perceived QoL after starting program was found and comparing anxious-depressive symptoms outcomes with control arm is still needed.
2016
- Clinical and molecular analysis of long-term HER2 positive metastatic breast cancer survivors.
[Poster]
Omarini, Claudia; Caprera, Cecilia; Manfredini, Samantha; Caggia, Federica; Guaitoli, Giorgia; Filieri, Maria Elisabetta; Moscetti, Luca; Bettelli, Stefania Raffaella; Kaleci, Shaniko; Cascinu, Stefano; Piacentini, Federico
abstract
ackground: Several multigene tests have been developed in Metastatic Breast Cancer (MBC) disease, in order to identify predictive factors correlated to clinical outcomes. The purpose of this study is to investigate the clinico-pathological and molecular characteristics that could differentiate long term responders from patients experiencing early progression during anti-HER2 treatments. Methods: A total of 34 HER2 positive MBC patients were included: 20 patients with a time to progression longer than 3 years in Long Responders group (LR) and 14 patients with a progression disease within one year of anti-HER2 therapy in Poor Responders group (PR). Tumor characteristics and treatment information were collected. The expression of 770 genes and 13 molecular pathways were evaluated using Nanostring PanCancer pathway panel performed on BC formalin-fixed paraffin-embedded tissues from diagnostic core biopsy or surgical resection. Results: Baseline patients and tumor characteristics were similar between the two groups, although PR patients were more likely to have CNS spread and more metastatic burden of disease compared to LR (29% vs. 0, p = 0.02 and 57% vs. 20%, p = 0.04, respectively). Gene expression analysis identified 30 genes with significantly different expression in the two cohorts; five of these were driver genes (BRCA1, PDGFRA, AR, PHF6 and MSH2). The majority of these genes were over-expressed, mainly in LR patients, and encoded growth factors, pro- or anti-inflammatory interleukins and DNA repair factors. Only four genes were down-regulated, all in PR group (TNFSF10, CACNG1, IL20RB and BRCA1). Most of these genes were involved in MAPK and PI3K pathways (9 and 8, respectively). MAPK pathway was differently expressed between LR and PR (p = 0.05). Even if not statistically significant but clinically relevant, PI3K was the only pathway overexpressed in PR patients (median expression LR: 1441 ± 485 vs 1759 ± 762 in PR group; p= 0.1). Conclusions: Whole genome expression analysis comparing LR vs. PR identified a group of genes that may predict more favourable long-term outcomes. Up-regulation of MAPK and down-regulation of PI3K pathways could be a positive predictive factors. Further clinical implications are warranted.
2016
- Molecular profile in primary and metastatic breast cancer treated with Exemestane and Everolimus.
[Abstract in Atti di Convegno]
Filieri, Maria Elisabetta; Bettelli, Stefania Raffaella; Maiorana, Antonio; Caprera, Cecilia; Manfredini, Samantha; Caggia, Federica; Iattoni, Elena; Cascinu, Stefano; Omarini, Claudia; Piacentini, Federico
abstract
Background. The PI3K/Akt/ mTOR pathway plays a significant role in endocrine resistance breast cancer (BC). Everolimus (EVE) has been approved after BOLERO-2 study, which showed a significant increase of PFS thanks to EVE plus Exemestane (EXE), compared to EXE alone. Hortobagyi et al. performed Next Generation Sequencing on 227 BOLERO-2 samples of primary BC to study the potential correlation between genetic alterations and EVE efficacy: a greater incidence of mutations in PI3KCA, PTEN, CCND1 and FGFR1 was detected, but treatment effect was independent from the genetic status. The aim of this study was to evaluate the molecular profile of both primary and secondary lesions in 25 metastatic BC patients treated with EVE+EXE in Modena University Hospital since 2014.
Materials and methods. Thirty-three DNA samples from 25 patients were examined, 13 from primary BC and 20 from metastatic lesions. In 8 patients both primary tumor and metachronous metastasis were evaluated. Genomic DNA samples from FFPE tissues was conducted using panel OncoCarta 2.0 on MassArray Sequenom platform that detect more than 150 single nucleotide variations from 18 genes (AKT1, BRAF, CTNNB1, FBX4, FBXW7, FGFR2/3, GNAQ, KIT, KRAS, MAP2K1/2, NRAS, PDGFR, PIK3CA, PTPN11, SOS1, TP53). Differences between mutational status of primary and metastatic BC have been evaluated using χ2 and Fisher tests.
Results. The median age was 54 years (range 50-67). 70% of patients had visceral involvement, 62% received more than 3 previous therapies, and for 8% of them an AI constituted the last treatment before EVE. Overall, 11 DNA samples were mutated (33%). 5 mutations were detected in the primary lesion with a frequency of 3% (PI3KCA, FBX4, KIT, MAP2K1, FBXW7) - 6 mutations in the metastasis (BRAF, KIT, TP53, FBXW7, CTNNB1, PI3KCA, AKT). Notably, mutations were found exclusively in primary lesion or in metastatic site, while only in one case both primary and secondary cancer were mutated, even if in two different genes. No significant correlation with treatment efficacy was evidenced.
Conclusions. The genes most frequently mutated in MBC were PI3KCA, AKT1 and FBXW7, even if the percentage of PIK3CA and AKT1 mutations was less than expected. No correlation between primary and metastatic mutational status was detected. Involving new patients maybe could be the way for more encouraging results.
2015
- Impact of time to surgery after neoadjuvant chemotherapy in patients with operable breast cancer.
[Abstract in Atti di Convegno]
Piacentini, F; Filieri, E; Grizzi, Giulia; Omarini, C; Maur, M; Guaitoli, G; Tazzioli, G; Madrigali, S; Caggia, F
abstract
Background: Some studies of adjuvant chemotherapy (CT) suggested that a shorter interval before the start of therapies may
improve survival outcomes in many groups of patients. Time to surgery (TTS) after neoadjuvant CT and survival outcomes have
not been established yet. The aim of this study is to evaluate the impact of TTS after neoadjuvant CT in terms of Overall Survival
(OS) and Disease Free Survival (DFS).
Patients and Methods: A retrospective analysis was conducted in 295 patients receiving neoadjuvant CT for stage I-IIIc breast
cancer between 1991 and 2013. 56 pts underwent surgery within 21 days (group A) from last CT cycle, 148 pts within 22-35 days
(group B) and 91 pts after 36 days (group C). The majority were infiltrating ductal carcinoma, stage IIA (37.6%) and IIB
(33.9%), with nodal involvement in 51.6% of the cases. LumA 18.3%, LumB/HER2- 28.2%, LumB/Her2+ 20.7%, HER2+ 9.8%,
TNBC 21%. All patients were treated with neoadjuvant CT: 70.5% with anthra-taxanes based regimen, 18% with anthra- alone,
10.9% with taxanes alone, 0.3% with CMF; plus Trastuzumab in 70% of HER2+ diseases.
Results: After a median follow up of 4.6 years, it was observed that patients in group A showed a significant better OS than
group B (HR 4.22; 95% CI, 1.27 – 14.00, p=0.018) and group C (HR 3.61; 95% CI, 1.01 – 12.86, p=0.048). Moreover group A
showed a significant better DFS than group B (HR 3.41; 95% CI 1.34 to 8.65, p=0.010) and group C (HR 3.77; 95% CI 1.42 to
9.95, p=0.007).
No correlations with OS were found in pts who achieved pCR (20.7%); pCR was predictive of better 5- and 10-years DFS
independently from TTS (95.4% in the pCR-group vs 75.4% of non-pCR group, HR 0.16; 95% CI 0.04 to 0.66, p=0.011). TTS
may influence DFS in non-pCR group: indeed 5-years DFS is 97.3% in group A, 72.7% in group B (HR 2.89; 95% CI 1.14 to
7.36, p=0.026), and 68.5% in group C (HR 3.44; 95% CI 1.3 to 9.1, p=0.013). No significant correlations with regard of stage at
diagnosis or molecular subtypes were found.
Conclusions: These results suggest that TTS after primary CT may influence patients' survival, regardless of stage at diagnosis
and tumor subtype, so that a shorter interval between that last cycle of neoadjuvant chemotherapy and breast surgery should be
addressed whenever possible .
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
- 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
- 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
- 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
- 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.