Transcriptional modulation of stress-related genes in association with early life stress exposure and trauma-focused psychotherapy in treatment resistant depression patients
This dataset contains mRNa expression data concerning the stress genes FKBP5, NR3C1 and SGK1 obtained in whole blood of patients suffering from treatment-resistant depression (TRD) with a history of early life stress undergoing an 8-week treatment with trauma-focused psychotherapy. B2M represents the housekeeping gene used for normalization. Ct, delta Ct (dCt) and 2^ -delta Ct (2^-dCt) values are reported. T0 = beginning of psychotherapy; T4 = 4 weeks of psychotherapy; T8 = end of psychotherapy; T12 = follow-up occurring 4 weeks after the end of psychotherapy. Percentage gene expression variations from T0 to T12 and from T0 to T8, calculated on the 2^-dCt values, are also shown. Rationale of the project and main findings: Introduction: Early life stress (ELS) associates with unfavourable treatment outcome in major depressive disorder and with Treatment-resistant depression (TRD), and trauma-focused psychotherapy is beneficial for TRD patients with ELS. Biological bases relating ELS and TRD lay on neuroendocrine and inflammatory dysregulations, mediated by Hypothalamic-Pituitary-Adrenal axis activated by stress. Objectives: We explored, in TRD patients, peripheral modulations of stress-response genes (NR3C1, FKBP5, SGK1) in relation to ELS and symptoms changes during psychotherapy and evaluated whether their baseline levels predict outcomes. Methods: 41 TRD patients participated. A subgroup (n=21) underwent trauma-focused psychotherapy. We used Montgomery-Åsberg Depression Rating Scale, Beck Depression Inventory-II and Beck Anxiety Inventory for symptoms evaluations, Childhood Experience of Care and Abuse Questionnaire for ELS assessment, and RT-qPCR for gene expression analysis at baseline (T0), 4 (T4) and 8 (T8) weeks of psychotherapy, and follow-up (T12). Results: We found higher NR3C1 and FKBP5 baseline levels in patients with mother neglect. From T0 to T8, we observed positive correlations of FKBP5, NR3C1 and SGK1 expression decrease with cognitive symptoms amelioration and of NR3C1 decrease with depressive symptoms improvement. Positive correlations of T0-T8 gene expression decrease with T0-T12 symptoms improvements were found, i.e., between FKBP5, NR3C1 and SGK1, and neurovegetative symptoms, between NR3C1 and SGK1, and self-reported depressive symptoms, and between SGK1 and anxiety symptoms. Baseline levels for all genes were higher in patients relapsing after 24 weeks. Conclusion: We found gene expression signatures of ELS. Amelioration during psychotherapy correlated with gene expression modulations, and transcripts baseline levels predicted relapse.
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Blood collection and mRNA isolation from blood Peripheral venous blood samples were collected in the morning between 8:00 and 9:00 a.m., after an overnight fast in PAXGene Blood RNA Tubes (Cat. 762165, Qiagen) for RNA isolation. PAXGene Blood RNA tubes were kept at room temperature for 2 hours, then frozen at -20 °C for 24 hours and finally stored at a -80 °C until RNA isolation. Total RNA was extracted from 2.5 mL of blood with the PAXGene Blood miRNA Kit (Cat. 763134, Qiagen), following manufacturer’s instructions. RNA quantification and quality control were carried out using spectrophotometric analysis (NanoDrop 2000, Thermo Scientific). Determination of candidate mRNA expression levels by RT-qPCR Expression levels of the target genes NR3C1 (Hs00353740_m1), FKBP5 (Hs01561006_m1), SGK1 (Hs00178612_m1) and of the housekeeping gene B2M (Hs99999907_m1) were analyzed using TaqMan Assays (Thermo Fisher, Waltham, MA, USA) and iTaq Universal Probes One-Step Kit (Bio-Rad Laboratories, Hercules, CA, USA) on the CFX384 Real Time PCR instrument (Bio-Rad Laboratories, Hercules, CA, USA) following manufacturer’s instructions. All Real-time PCR reactions were carried out with the following steps: 10 minutes at 50°C, 5 minutes at 95°C followed by 39 cycles of 10 seconds at 95°C and 30 seconds at 60°C. All reactions were run in triplicate. The Ct values were normalized according to ΔCt method on the housekeeping gene B2M, which was stably expressed across samples.