Reproductive Endocrine Related Mood Disorders-Differential Sensitivity

生殖内分泌相关情绪障碍-敏感性差异

• 批准号: 10266604
• 负责人: Peter Schmidt
• 金额: $ 66.33万
• 依托单位: NATIONAL INSTITUTE OF MENTAL HEALTH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10266604
• 关键词: Accounting; Affective; Agonist; Allopregnanolone; Anterior; Appearance; Attention; Behavioral; Biological; Birth; Brain; Calcium Signaling; Cell Culture Techniques; Cell Line; Cerebrovascular Circulation; Childbirth; Chronic Disease; Clinical Protocols; Clinical Trials; Collaborations; Complex; Data; Development; Diagnosis; Direct Costs; Disease; Disease remission; Dissociation; Dutasteride; Endocrine; Endoplasmic Reticulum; Enzyme Inhibitor Drugs; Epigenetic Process; Estradiol; Event; Facilities and Administrative Costs; Functional disorder; Future; GNRH1 gene; Gene Expression; Gene Family; Gene Silencing; Genes; Genetic; Genetic Transcription; Golgi Apparatus; Gonadal Steroid Hormones; Health; Homeostasis; Hormonal; Hormone Receptor; Hormone use; Hormones; Human; Hypothyroidism; In Vitro; Infant; Intervention; Investigation; Lead; Linear Models; Longevity; Luteal Phase; Major Depressive Disorder; Measures; Mediating; Menstrual cycle; Mental Depression; MicroRNAs; Mood Disorders; Morbidity - disease rate; National Institute on Alcohol Abuse and Alcoholism; Nature; Neurons; Neurotransmitters; Ovarian; Ovarian Ablation; Oxidoreductase; Pathway Analysis; Pathway interactions; Pattern; Performance; Perimenopause; Pharmaceutical Preparations; Phenotype; Physiological; Placebos; Postpartum Depression; Postpartum Period; Predisposition; Pregnancy; Premenstrual syndrome; Production; Progesterone; Proteins; Protocols documentation; Psychotic Disorders; Puerperium; Quantitative Reverse Transcriptase PCR; RNA Splicing; Regulation; Reporting; Reproductive Periods; Research Design; Rest; Risk; Role; Series; Signal Transduction; Small RNA; Source; Statistical Models; Steroid Receptors; Steroids; Stimulus; Symptoms; System; Testing; Thapsigargin; Tissues; Translating; Validation; Vascular Endothelial Growth Factors; Withdrawal; Woman; Women' s Group; Work; XBP1 gene; behavioral outcome; behavioral response; biological adaptation to stress; burden of illness; case control; cingulate cortex; clinical Diagnosis; cognitive function; depressive symptoms; differential expression; endoplasmic reticulum stress; functional genomics; hormone sensitivity; lymphoblastoid cell line; mRNA Expression; meetings; member; mood symptom; mortality; neuroimaging; neuroregulation; neurosteroids; novel therapeutics; overexpression; premenstrual dysphoric disorder; prevent; protein expression; puberty transition; relating to nervous system; reproductive; response; therapy development; transcriptome; transcriptome sequencing; transcriptomics; treatment trial

This report includes work arising from the following clinical protocols: NCT00005011, NCT00056901, NCT00059228, NCT00082043, NCT00100360, NCT00001177, NCT00001259, and NCT00001481. Our studies have documented that PMDD symptoms are eliminated by ovarian suppression and stimulated by administration of ovarian steroids, yet appear in the context of levels of ovarian steroids indistinguishable from those in women without PMDD. Additionally, we demonstrated that the change in levels of combined estradiol and progesterone from low to high, and not the steady state levels, triggered the onset of PMDD symptoms in women with this condition whose PMDD was in remission after GnRH agonist-induced ovarian suppression. Our findings provide a new target on which interventions could be focused (namely increasing neurosteroid levels from the follicular to the luteal phase). We will conduct a placebo-controlled treatment trial investigating the effects of stabilizing neurosteroid levels with dutasteride (a 5alpha reductase enzyme inhibitor which prevents the production of the neurosteroid, allopregnanolone from progesterone that is implicated in the alterations of neurotransmitter function that could lead to PMDD) in women with PMDD. We also employed our ovarian steroid manipulation studies (GnRH agonist studies) to explore possible neural substrates of risk in women with PMDD. We have identified the subgenual anterior cingulate cortex (SGCC) to be differentially regulated by ovarian steroids in women with PMDD compared with control women (i.e., regional cerebral blood flow rCBF during a resting state exam is decreased during estradiol or progesterone exposure when PMDD symptoms recur). The degree of altered rCBF also correlated with gene expression in the ESC/E(Z) pathway of genes a family of genes which we previously identified to be differentially expressed (increased) in cell lines from women with PMDD compared to controls, and members of which are differentially regulated by estradiol and progesterone in cell culture (see below). Additionally, the results of a whole transcriptome sequencing study of the SGCC shows evidence that sex steroid receptors are present in this tissue and, therefore, it is plausible that SGCC activity could be regulated by ovarian steroids. These data suggest both the relevance of differences in the response of the SGCC to ovarian steroids with consequent differential activation of brain networks that mediate the affective responsivity of women with PMDD as well as providing a biologically plausible target for neuromodulatory interventions in PMDD. These behavioral and neuroimaging data also set the stage for the performance of in vitro cellular studies of ovarian steroid responsivity in women with PMDD. In collaboration with Dr. David Goldman at NIAAA, we developed lymphoblastoid cell lines (LCLs) and human induced-pluripotent cell lines (h-IPSCs) from women with and without PMDD who had participated in our GnRH agonist-induced ovarian suppression studies. The behavioral outcomes observed during these protocols serve to refine the hormone-sensitive phenotype beyond simply the established clinical diagnoses. Our in vitro experimental strategies attempt to recapitulate the endocrine events that trigger mood symptoms in women with PMDD. In our first study, pathway analyses of the LCL transcriptome revealed, among others, over-expression of ESC/E(Z) complex genes (an ovarian steroid-regulated gene silencing complex) in untreated LCLs from women with PMDD, with more than half of these genes over-expressed as compared to controls. In contrast, protein expression of ESC/E(Z) genes was decreased in untreated PMDD LCLs. Finally, mRNA expression of several ESC/E(Z) complex genes were increased by P in controls only and decreased by E in PMDD LCLs. These findings provided the first evidence of a plausible biological substrate for the differential behavioral response to E/P in women with PMDD. Indeed, these data suggest that women with PMDD have an intrinsic abnormality in their epigenetic capacity that could manifest in an alteration in their ability to translate environmental events into long-term changes in gene expression. We have continued to pursue these findings in PMDD. During the past year we have pursued both the mechanism underlying the altered expression of the ESC/E(Z) pathway and the effects of ovarian steroid exposures on gene expression in PMDD versus controls. First, preliminary evidence from a whole small RNA sequencing (targeting micro-RNA expression) shows a significantly increased expression of several micro-RNAs in LCLs from women with PMDD compared to those in controls. Additionally, several of these differentially expressed micro-RNAs target the ESC/E(Z) pathway, and, therefore, could contribute to the observed dissociation between increased expression of the ESC/E(Z) pathway genes and decreased protein levels of this same pathway in PMDD. Additionally, several of these micro-RNAs target the other genes relevant for both affective dysregulation and tissue-specific steroid signaling including vascular endothelial growth factor (VEGF). Second, we performed transcriptomic analyses of LCLs derived from women with PMDD and asymptomatic controls cultured under untreated (steroid-free), estradiol-treated, and progesterone-treated conditions. Weighted gene correlation network analysis (WGCNA) of transcriptomes identified four gene modules with significant differences in the response patterns in women with PMDD (versus control LCLs) that also differed across hormone exposures, including one enriched for neuronal functions. Exploratory enrichment analysis of hub genes underlying neuronal enrichment signals revealed multiple pathways governing intracellular Ca2+ dynamics. Next, in a gene-level analysis comparing transcriptional response to hormone across diagnoses, a statistical model (i.e., generalized linear model) identified 1522 genes differentially responsive to estradiol (E2-DRGs). Among the top 10 E2-DRGs was an interacting gene network (NUCB1, DST, GCC2, GOLGB1) involved in endoplasmic reticulum (ER)-Golgi function. qRT-PCR validation reproduced a diagnosis by hormone interaction (reflecting differential expression patterns in LCLs from women with PMDD compared with those from controls during estradiol exposure), for NUCB1, a regulator of cellular Ca2+ and ER stress. Finally, we used a thapsigargin (Tg) challenge to test whether estradiol induces differences in Ca2+ homeostasis and ER stress response in PMDD. Untreated PMDD LCLs had a 1.36-fold decrease in Tg-induced XBP1 splicing response (a measure of intracellular calcium signaling) compared to controls, and a 1.62-fold decreased response in the E2-exposed condition. These data suggest that estradiol-dependent aberrations in cellular Ca2+ dynamics and ER stress may contribute to the pathophysiology of PMDD and could guide treatment development to focus on medications regulating intracellular calcium signaling (known to be involved with affective state regulation) in PMDD. In a parallel series of functional genomic studies employing LCLs and h-IPSCs developed from women with PPD and in a separate group of women with first-onset postpartum psychosis (PPP) we have completed whole transcriptome sequencing. Whole transcriptome RNA-sequencing is completed in LCLs from controls and women with PPD under conditions of untreated baseline, supraphysiologic (high) combined estradiol and progesterone (replicating pregnancy), and hormone withdrawal (replicating hormonal events of parturition). Preliminary findings revealed multiple differentially expressed genes in all conditions meeting FDR corrections, particularly in the high combined estradiol and progesterone conditions between cases and controls.

本报告包括以下临床方案的工作：NCT00005011、NCT00056901、NCT00059228、NCT00082043、NCT00100360、NCT00001177、NCT00001259 和 NCT00001481。 我们的研究证明，经前抑郁症的症状可以通过卵巢抑制和卵巢类固醇的刺激来消除，但其卵巢类固醇水平与没有经前抑郁症的女性没有什么区别。此外，我们证明，雌二醇和黄体酮组合水平从低到高的变化，而不是稳态水平的变化，引发了患有这种疾病的女性经前抑郁症症状的发作，这些女性在 GnRH 激动剂诱导的卵巢抑制后，经前抑郁症已得到缓解。我们的研究结果提供了一个可以集中干预的新目标（即增加从卵泡到黄体期的神经类固醇水平）。我们将进行一项安慰剂对照治疗试验，研究度他雄胺（一种 5α 还原酶抑制剂，可防止黄体酮产生神经类固醇、四氢孕酮，而度他雄胺与神经递质功能的改变有关，从而可能导致经前抑郁症）稳定神经类固醇水平的效果。患有经前抑郁症 (PMDD) 的女性。 我们还利用卵巢类固醇操作研究（GnRH 激动剂研究）来探索患有经前抑郁症的女性的风险的可能神经基础。我们发现，与对照女性相比，患有经前抑郁症的女性膝下前扣带皮层 (SGCC) 受到卵巢类固醇的差异性调节（即，当经前抑郁症症状复发时，静息状态检查期间的局部脑血流量 rCBF 在雌二醇或黄体酮暴露期间减少）。 rCBF 改变的程度还与 ESC/E(Z) 通路中基因家族的基因表达相关，我们之前发现该基因家族在患有 PMDD 的女性的细胞系中与对照组和它们在细胞培养物中受到雌二醇和孕酮的差异调节（见下文）。 此外，SGCC 的全转录组测序研究结果表明，该组织中存在性类固醇受体，因此 SGCC 活性可能受到卵巢类固醇的调节是合理的。这些数据表明，SGCC 对卵巢类固醇反应的差异与随后介导经前抑郁症女性情感反应的大脑网络的差异激活有关，并为经前抑郁症的神经调节干预提供了生物学上合理的目标。 这些行为和神经影像数据也为患有经前抑郁症的女性卵巢类固醇反应性的体外细胞研究奠定了基础。我们与 NIAAA 的 David Goldman 博士合作，从参加过 GnRH 激动剂诱导的卵巢抑制研究的患有或未患有经前抑郁症的女性中开发了类淋巴母细胞系 (LCL) 和人诱导多能细胞系 (h-IPSC)。在这些方案中观察到的行为结果有助于完善激素敏感表型，而不仅仅是既定的临床诊断。我们的体外实验策略试图重现引发经前抑郁症女性情绪症状的内分泌事件。在我们的第一项研究中，LCL 转录组的通路分析显示，除其他外，在未经治疗的经前抑郁症 (PMDD) 女性 LCL 中，ESC/E(Z) 复合体基因（一种卵巢类固醇调节基因沉默复合体）过度表达，其中一半以上与对照相比，这些基因的过度表达。相比之下，未经处理的 PMDD LCL 中 ESC/E(Z) 基因的蛋白表达降低。最后，仅在对照中，几个 ESC/E(Z) 复合体基因的 mRNA 表达因 P 增加而在 PMDD LCL 中因 E 降低。这些发现为患有经前抑郁症 (PMDD) 的女性对 E/P 的不同行为反应提供了合理的生物学基础的第一个证据。事实上，这些数据表明，患有经前抑郁症的女性的表观遗传能力存在内在异常，这可能表现为她们将环境事件转化为基因表达长期变化的能力发生改变。我们继续在经前抑郁症 (PMDD) 中研究这些发现。在过去的一年中，我们研究了 ESC/E(Z) 通路表达改变的潜在机制，以及卵巢类固醇暴露对 PMDD 与对照基因表达的影响。首先，来自全小 RNA 测序（针对 micro-RNA 表达）的初步证据表明，与对照组相比，患有 PMDD 的女性 LCL 中几种 micro-RNA 的表达显着增加。此外，其中一些差异表达的 micro-RNA 靶向 ESC/E(Z) 通路，因此可能有助于观察到 ESC/E(Z) 通路基因表达增加与该通路基因蛋白水平降低之间的分离。 PMDD 的途径。此外，其中一些 micro-RNA 靶向与情感失调和组织特异性类固醇信号传导相关的其他基因，包括血管内皮生长因子 (VEGF)。 其次，我们对来自患有经前抑郁症 (PMDD) 的女性和在未治疗（无类固醇）、雌二醇治疗和黄体酮治疗条件下培养的无症状对照的 LCL 进行了转录组分析。转录组的加权基因相关网络分析 (WGCNA) 发现了四个基因模块，这些模块在患有经前抑郁症 (PMDD) 的女性（与对照 LCL）中的反应模式存在显着差异，这些模块在不同激素暴露情况下也存在差异，其中包括一种富含神经元功能的基因模块。对神经元富集信号背后的中枢基因的探索性富集分析揭示了控制细胞内 Ca2+ 动态的多种途径。接下来，在比较不同诊断中对激素的转录反应的基因水平分析中，统计模型（即广义线性模型）确定了 1522 个对雌二醇（E2-DRG）有差异反应的基因。排名前 10 位的 E2-DRG 包括参与内质网 (ER)-高尔基体功能的相互作用基因网络（NUCB1、DST、GCC2、GOLGB1）。 qRT-PCR 验证通过激素相互作用再现了 NUCB1（细胞 Ca2+ 和 ER 应激调节因子）的诊断（反映了经前抑郁症 (PMDD) 女性与对照组在雌二醇暴露期间 LCL 中的差异表达模式）。最后，我们使用毒胡萝卜素 (Tg) 激发来测试雌二醇是否会引起经前抑郁症 (PMDD) 中 Ca2+ 稳态和 ER 应激反应的差异。与对照组相比，未经处理的 PMDD LCL 的 Tg 诱导的 XBP1 剪接反应（细胞内钙信号传导的测量）降低了 1.36 倍，并且在 E2 暴露条件下的反应降低了 1.62 倍。这些数据表明，细胞 Ca2+ 动力学和 ER 应激中的雌二醇依赖性畸变可能导致 PMDD 的病理生理学，并可以指导治疗开发重点关注调节 PMDD 细胞内钙信号传导（已知与情感状态调节有关）的药物。 在一系列平行的功能基因组研究中，我们使用从患有 PPD 的女性和另一组患有首次发作的产后精神病 (PPP) 的女性中开发的 LCL 和 h-IPSC 进行了全转录组测序。在未经治疗的基线、超生理（高）雌二醇和黄体酮组合（复制妊娠）和激素戒断（复制分娩的激素事件）的条件下，对对照和患有 PPD 的女性的 LCL 进行全转录组 RNA 测序。初步研究结果显示，在满足 FDR 校正的所有条件下，特别是在病例和对照之间雌二醇和黄体酮组合较高的条件下，存在多个差异表达基因。