The Neuroregulatory Effects of Gonadal Steroids in Humans

性腺类固醇对人类的神经调节作用

• 批准号: 10011366
• 负责人: Peter Schmidt
• 金额: $ 71.96万
• 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/10011366
• 关键词: Acute; Adrenergic Agents; Affect; Affective; Agonist; Amygdaloid structure; BDNF gene; Back; Behavior; Behavioral; Biological; Brain; Brain region; Brain-Derived Neurotrophic Factor; Catechol O-Methyltransferase; Cell Line; Cells; Cerebrovascular Circulation; Clinical Protocols; Code; Cognitive; Collaborations; Companions; Complex; Data; Disease remission; Dopamine; Estradiol; Estrogens; Female; Functional Magnetic Resonance Imaging; Functional disorder; Gene Expression; Gene Silencing; Generations; Genes; Genetic; Genetic Variation; Genotype; Goals; Gonadal Steroid Hormones; Gonadotropin Hormone Releasing Hormone; Heterozygote; Hippocampus (Brain); Hormonal; Hormones; Human; Hypogonadism; Immune; Individual; Insula of Reil; Measures; Medial; Mediating; Memory; Menstrual cycle; Mood Disorders; Moods; Mus; Neurons; Ovarian; Ovarian Ablation; Ovarian hormone; Ovariectomy; Overlapping Genes; Oxygen; Pathway interactions; Phenotype; Physiological; Positron-Emission Tomography; Prefrontal Cortex; Premenopause; Procedures; Progesterone; Promoter Regions; Protocols documentation; Recurrence; Regulation; Reporting; Response Elements; Rest; Rewards; Role; Sex Differences; Short-Term Memory; Stains; Steroid Receptors; Steroids; Stress; Symptoms; System; Techniques; Testing; Time; Tissues; Undifferentiated; Universities; Variant; Water; Wild Type Mouse; Wisconsin; Withdrawal; Woman; Work; anxiety-like behavior; behavioral response; cingulate cortex; cognitive testing; disorder control; dopaminergic neuron; experience; functional MRI scan; functional genomics; genetic variant; genomic variation; hormone sensitivity; hypothalamic-pituitary-adrenal axis; induced pluripotent stem cell; lymphoblastoid cell line; nerve stem cell; neural circuit; neuroimaging; neurophysiology; neurosteroids; performance tests; phenomenological models; precursor cell; premenstrual dysphoric disorder; relating to nervous system; response; reward processing; serotonin transporter; social; trafficking; transcriptome; transcriptome sequencing

This report includes work arising from the following clinical protocols: NCT00026832, NCT00100360, NCT00001177, and NCT00001322. Previous studies have identified the presence of abnormalities in resting state activity and reward processing in mood disorders. Indeed, altered resting state function represents an intriguing possible explanation for the generation of ovarian hormone-related state changes characterized by predictable alterations in social, cognitive (e.g., autobiographical memory), and affective function in women with PMDD. Results from our resting state H215O PET studies in healthy premenopausal women (n = 40) demonstrate effects of both hypogonadism and ovarian steroids on resting rCBF in several brain regions, including BA25, insula, and medial PFC (mPFC). Evidence also suggests that genomic variation in sex-steroid receptors or in the systems regulated by sex steroids may contribute to the ovarian hormone-sensitivity observed in PMDD. Thus, as a first step, we examined the effects of common genetic variations in women during each of the hormonal conditions established within the GnRH agonist-induced hypogonadism and ovarian steroid addback protocol. We have tested specific, common functional variants (BDNF Val66Met and COMT Val158Met) that are regulated by sex steroids and that are known to alter brain function. In our studies, therefore, we employ these gene variants as probes for the impact of genetic substrates on the effects of ovarian steroids on womens brain function. We have previously identified that variations in the BDNF gene, which is modulated by ovarian hormones and contains an estrogen response element (ERE), differentially affect working memory-related functions only in the presence of estradiol and only in the hippocampus in healthy women (Wei et al. 2018). Similarly, the gene coding for Catechol-O-Methyltransferase (COMT) contains both EREs and progesterone response elements (PREs) in its promoter region, its expression in brain can be regulated by estradiol (and in some tissues by progesterone) and COMT activity is increased by circulating estradiol in women. Indeed, two studies in women reported the modulatory effects on working memory-related brain regions of both estradiol and COMT genotype (Dumas et al. 2018, Jacobs and D'Esposito 2011). To further understand the interactions between ovarian steroids and BDNF/COMT genotypes on brain circuits related to hippocampal and PFC activity, we used oxygen-15 water regional cerebral blood flow (rCBF) positron emission tomography (PET) to study healthy women who participated the GnRH agonist hormone manipulation study. Preliminary findings demonstrate an ovarian hormone-by-genotype interaction in the hubs of the working memory network with BDNF genotype and estradiol regulating activity in hippocampus, and COMT genotype and estradiol regulating activity within the PFC. These data suggest that the effects of sex hormones on working memory-related brain function are mediated by genotypic variations in a brain region-specific manner. In a companion collaboration focusing on the BDNF gene with the Rockefeller University, we examine the effects of ovariectomy and estradiol replacement on both behavior and hippocampal gene expression in the female humanized BDNF heterozygous Met mouse. We observed that heterozygous Met mice show a differential behavioral sensitivity to estradiol in several hippocampal tasks and in anxiety-like behaviors. Additionally, whole transcriptome RNA sequencing in the BDNF heterozygous Met mouse (hippocampus) and in women with PMDD (lymphoblastoid cell lines) show a significant overlap in the expression of several gene networks stimulated by estradiol. No similar overlap in gene expression was observed after estradiol exposure in either wild type mice or asymptomatic control women. This cross-species overlap in gene expression suggests the potential physiologic (and behavioral) relevance of the BDNF heterozygote met mouse to our understanding of PMDD. Overall, the findings from these studies demonstrate that in healthy women, variations in genes relevant to ovarian steroids can impact functional neurocircuitry and network level gene expression in regionally- and hormonally-specific ways. Specifically, interactions between BDNF and COMT genotypes and estradiol on the activation of functional neurocircuitry in the hippocampus (where BDNF is highly expressed) and the DLPFC (where COMT has primacy for dopamine trafficking), respectively. In studies currently underway, we will examine these gene-hormone interactions in reward-responsive and resting state neural circuits. Our functional genomic studies in cell lines from both women with and without PMDD have demonstrated the relevance of our prior finding of abnormal function of the ESC/E(Z) gene complex in PMDD as follows: First, neuroprogenitor cells (NPCs) were successfully differentiated from induced pluripotent stem cells (iPSCs) made from women with PMDD and control women (confirmed by immunofluorescent staining and transcriptome analysis). Overlapping gene expression between lymphoblastoid cells (LCLs) and NPCs reveals that several genes and gene pathways potentially important for PMDD pathophysiology are expressed in both. Indeed, both LCLs and NSCs show signatures of differential gene silencing through similar mechanisms at baseline and in response to ovarian steroids. These findings in NSCs reveal neuronal differences between women with PMDD and controls both at baseline and after ovarian steroids, including ESC/E(Z) targets. Second, we employed two stock neuronal cell lines: Luhmes Cells, which are immortalized neuronal precursor cell lines (both undifferentiated/neuronal precursors and differentiated/mature dopaminergic neurons), as well as SH-SY5Y, an immortalized neuroblastoid cell line characterized by an adrenergic and dopaminergic phenotype. Both stock neuronal cell lines contained evidence of ESC/E(Z) gene complex function and responsivity to ovarian steroids. Thus, these data provide strong supportive evidence of the neural relevance of our initial findings in lymphoblastoid cell lines (immune-origin tissue) in several more pathophysiologically-relevant tissues (i.e., human NPCs, two stock neuronal cell lines).

本报告包括以下临床方案的工作：NCT00026832、NCT00100360、NCT00001177 和 NCT00001322。 先前的研究已经发现情绪障碍中静息状态活动和奖赏处理存在异常。事实上，静息状态功能的改变为卵巢激素相关状态变化的产生提供了一个有趣的可能解释，其特征是经前抑郁症女性的社交、认知（例如自传体记忆）和情感功能的可预测改变。我们对健康绝经前女性 (n = 40) 进行静息态 H215O PET 研究的结果表明，性腺功能减退症和卵巢类固醇对多个大脑区域（包括 BA25、岛叶和内侧 PFC (mPFC)）的静息 rCBF 都有影响。 证据还表明，性类固醇受体或性类固醇调节系统的基因组变异可能导致经前抑郁症中观察到的卵巢激素敏感性。因此，作为第一步，我们检查了在 GnRH 激动剂诱导的性腺功能减退症和卵巢类固醇加回方案中建立的每种激素条件下，女性常见遗传变异的影响。我们测试了特定的常见功能变体（BDNF Val66Met 和 COMT Val158Met），这些变体受性类固醇调节，并且已知会改变大脑功能。因此，在我们的研究中，我们利用这些基因变异作为探针，研究遗传底物对卵巢类固醇对女性大脑功能影响的影响。 我们之前已经发现，受卵巢激素调节并含有雌激素反应元件（ERE）的 BDNF 基因的变异，仅在存在雌二醇的情况下以及仅在健康女性的海马体中对工作记忆相关功能产生差异性影响（Wei et al. 2018）。类似地，编码儿茶酚-O-甲基转移酶（COMT）的基因在其启动子区域同时包含ERE和孕酮反应元件（PRE），其在大脑中的表达可以通过雌二醇（在某些组织中通过孕酮）调节，并且COMT活性通过女性循环中的雌二醇而增加。事实上，两项针对女性的研究报告了雌二醇和 COMT 基因型对工作记忆相关大脑区域的调节作用（Dumas 等人，2018 年；Jacobs 和 D'Esposito，2011 年）。 为了进一步了解卵巢类固醇和 BDNF/COMT 基因型对海马和 PFC 活动相关脑回路的相互作用，我们使用氧 15 水局部脑血流 (rCBF) 正电子发射断层扫描 (PET) 对参与 GnRH 激动剂激素操纵研究的健康女性进行研究。初步研究结果表明，工作记忆网络中枢的卵巢激素与基因型之间存在相互作用，其中包括海马中的 BDNF 基因型和雌二醇调节活性，以及​​ PFC 中的 COMT 基因型和雌二醇调节活性。 这些数据表明，性激素对工作记忆相关大脑功能的影响是由大脑区域特定方式的基因型变异介导的。 在与洛克菲勒大学专注于 BDNF 基因的合作中，我们研究了卵巢切除和雌二醇替代对雌性人源化 BDNF 杂合 Met 小鼠行为和海马基因表达的影响。我们观察到杂合 Met 小鼠在一些海马任务和焦虑样行为中对雌二醇表现出不同的行为敏感性。此外，BDNF 杂合 Met 小鼠（海马）和患有 PMDD 的女性（淋巴母细胞系）的全转录组 RNA 测序显示，雌二醇刺激的几个基因网络的表达存在显着重叠。在野生型小鼠或无症状对照女性中暴露于雌二醇后，没有观察到类似的基因表达重叠。这种基因表达的跨物种重叠表明 BDNF 杂合子小鼠与我们对 PMDD 的理解具有潜在的生理（和行为）相关性。 总体而言，这些研究的结果表明，在健康女性中，与卵巢类固醇相关的基因变异可以以区域和激素特异性方式影响功能神经回路和网络水平基因表达。具体来说，BDNF 和 COMT 基因型与雌二醇之间的相互作用分别对海马体（BDNF 高度表达）和 DLPFC（COMT 在多巴胺运输中起主要作用）的功能性神经回路的激活产生影响。在目前正在进行的研究中，我们将检查奖励反应和静息状态神经回路中的这些基因-激素相互作用。 我们对患有和不患有经前抑郁症 (PMDD) 女性的细胞系进行的功能基因组研究证明了我们之前发现的 ESC/E(Z) 基因复合物在经前抑郁症 (PMDD) 中的异常功能的相关性，如下所示：首先，神经祖细胞 (NPC) 成功地从来自患有经前抑郁症 (PMDD) 女性和对照女性的诱导多能干细胞 (iPSC) 中分化出来（通过免疫荧光染色和转录组证实） 分析）。类淋巴母细胞 (LCL) 和 NPC 之间的基因表达重叠表明，对 PMDD 病理生理学潜在重要的几个基因和基因通路在两者中均表达。事实上，LCL 和 NSC 都通过相似的基线机制和对卵巢类固醇的反应显示出差异基因沉默的特征。 NSC 的这些发现揭示了患有 PMDD 的女性和对照组女性在基线和卵巢类固醇治疗后的神经元差异，包括 ESC/E(Z) 目标。其次，我们采用了两种库存神经元细胞系：Luhmes细胞，其是永生化神经元前体细胞系（未分化/神经元前体和分化/成熟多巴胺能神经元），以及SH-SY5Y，其是一种以肾上腺素能和多巴胺能表型为特征的永生化神经母细胞系。两种储备神经元细胞系均含有 ESC/E(Z) 基因复合体功能和对卵巢类固醇反应性的证据。因此，这些数据为我们在几种病理生理学相关组织（即人类 NPC，两种库存神经元细胞系）中的类淋巴母细胞系（免疫源组织）中的初步发现的神经相关性提供了强有力的支持证据。