Dysregulation of HPA-axis genes in Bipolar Disorder

双相情感障碍中 HPA 轴基因的失调

• 批准号: 8770987
• 负责人: Richard S Lee
• 金额: $ 30.22万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-11 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8770987
• 关键词: Adopted; Adoption; Adverse effects; Alternative Splicing; Amygdaloid structure; Animal Model; Autopsy; BAG1 gene; Behavioral; Biological Assay; Biology; Bipolar Disorder; Brain; Brain-Derived Neurotrophic Factor; CRH gene; Candidate Disease Gene; Clinical; Complex; Control Groups; Custom; DNA; DNA Methylation; Data; Development; Disease; Drug usage; Early-life trauma; Enhancers; Environmental Risk Factor; Epidemiology; Epigenetic Process; Etiology; Event; Exons; Exposure to; Family Study; Fluorescence; Fluorescence-Activated Cell Sorting; Future; Gene Expression; Gene Proteins; Genes; Genetic; Genetic Risk; Genomic DNA; Glucocorticoids; Gold; Heat-Shock Proteins 70; Heat-Shock Proteins 90; Hepatotoxicity; Hippocampus (Brain); Human; Individual; Institutes; Introns; Investigation; Knowledge; Lead; Link; Literature; Manic; Measures; Mediating; Mental Depression; Mental disorders; Messenger RNA; Metabolic syndrome; Methylation; Molecular; Molecular Biology; Mood Disorders; Moods; Neurons; Pathogenesis; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Post-Translational Protein Processing; Prefrontal Cortex; Process; Protein Isoforms; Proteins; Public Health; RNA; Recording of previous events; Recurrence; Regulation; Relapse; Risk Factors; Role; Sampling; Scanning; Signal Transduction; Societies; Sorting - Cell Movement; Stress; Symptoms; System; Techniques; Tissues; Transcript; Twin Multiple Birth; Validation; Western Blotting; biological adaptation to stress; bisulfite; brain tissue; burden of illness; cohort; disorder control; fighting; gene function; hypothalamic-pituitary-adrenal axis; improved; innovation; mRNA Expression; neuropsychiatry; next generation sequencing; prevent; public health relevance; pyrosequencing; research study; response; tacrolimus binding protein 4

DESCRIPTION (provided by applicant): Bipolar disorder (BP) poses a significant disease burden on society, due to our lack of knowledge of its etiology and mechanism of action of drugs that are used to treat it. Genetic, clinical, and epidemiological evidence suggests the presence of a complex interplay of genetic and environmental risk factors in bipolar disorder. In particular, the HPA-axis system responsible for mounting the "fight-or-flight" response may constitute a significant environmental factor that plays a role in precipitating and exacerbating bipolar disorder symptoms. In order to demonstrate the role of HPA-axis genes in bipolar disorder, we propose to examine in careful detail a set of genes that are: directly involved in mediating the stress-response; implicated in neuropsychiatric disorders or behavioral response to stress; or targeted by or directly influence glucocorticoid-signaling. Our proposal draws strength from preliminary data that show alterations in DNA methylation and expression of the stress-response gene FKBP5 in postmortem brains of bipolar patients. In Aim 1, we will first obtain postmortem brain tissues from control and bipolar samples (N=25 each) that have been well defined in terms of demographic information and drug history. We will process these brain tissues by fluorescence-activated cell sorting (FACS), in order to obtain a more homogeneous neuronal fraction. We will extract genomic DNA from the neuronal fractions, and, using the Methyl-Seq platform, enrich for candidate HPA-axis genes that have been implicated in mood disorders and stress-induced behavioral deficits in animal models. Enriched DNA will then be bisulfite-converted and subjected to next-generation sequencing. DNA methylation data for the candidate HPA-axis genes will be analyzed to identify differentially methylated regions (DMRs) between BP and control tissues. Further, DMRs will be independently verified using pyrosequencing, considered the gold standard for DNA methylation analysis. In Aim 2, we will extract messenger RNA from unsorted tissues to assess expression levels of the candidate HPA-axis genes, using multiple Taqman probes to target isoform-specific exon-intron junctions. We will also perform Western blotting on proteins extracted from the brain tissues to determine the concordance between gene expression and protein levels in bipolar vs. control samples. Results from Aims 1 and 2 will be further analyzed to identify potential DNA methylation-independent mechanisms, alternative splicing, and post-translational modification events specific to bipolar disorder. Taken together, the proposed experiments seek to demonstrate the role that HPA-axis genes may play in bipolar disorder pathology and to provide additional avenues for future investigation into differential regulation of HPA-axis substrates in bipolar disorder.

描述（由申请人提供）：双相情感障碍（BP）对社会造成了重大疾病负担，因为我们对其病因和用于治疗它的药物的病因和作用机理缺乏知识。遗传，临床和流行病学证据表明，躁郁症中存在遗传和环境风险因素的复杂相互作用。特别是，负责安装“战斗或飞行”响应的HPA轴系统可能构成一个重要的环境因素，该因素在促进和加剧双相情感障碍症状方面发挥作用。为了证明HPA轴基因在双相情感障碍中的作用，我们建议在仔细详细详细检查一组基因，这些基因直接参与介导应激反应；与神经精神疾病或对压力的行为反应有关；或由或直接影响糖皮质激素信号。我们的建议从初步数据中获取强度，这些数据显示了双极患者后大脑中DNA甲基化和应力反应基因FKBP5的表达改变。在AIM 1中，我们将首先从对照和双极样品（n = 25）中获得验尸后脑组织，这些样品已在人口统计学信息和药物病史方面得到很好的定义。我们将通过荧光激活的细胞分选（FACS）来处理这些脑组织，以获得更均匀的神经元分数。我们将从神经元分数中提取基因组DNA，并使用甲基序列平台富含与情绪障碍和动物模型中压力诱导的行为缺陷有关的候选HPA轴基因。然后将富集的DNA转换并进行下一代测序。将分析候选HPA轴基因的DNA甲基化数据，以鉴定BP和对照组织之间的差异甲基化区域（DMR）。此外，DMR将使用焦磷酸测序（被认为是DNA甲基化分析的金标准）独立验证。在AIM 2中，我们将使用多个TAQMAN探针靶向同工型特异性外显子 - 内隆连接点，从未分类的组织中提取信使RNA，以评估候选HPA轴基因的表达水平。我们还将对从脑组织提取的蛋白质进行蛋白质印迹，以确定双极与对照样品中基因表达与蛋白水平之间的一致性。目标1和2的结果将进一步分析，以确定潜在的DNA甲基化机制，替代剪接以及针对双相情感障碍的翻译后修饰事件。综上所述，拟议的实验试图证明HPA轴基因在双相情感障碍病理学中可能起的作用，并为将来研究双相情感障碍中HPA轴底物的差异调节提供了其他途径。