Novel Role for Sirtuin Signaling Mechanisms and Downstream Targets in Depression

Sirtuin 信号机制和下游靶点在抑郁症中的新作用

• 批准号: 8440735
• 负责人: Deveroux Ferguson
• 金额: $ 8.99万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-07 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8440735
• 关键词: ADP Ribose Transferases; Affect; Animal Model; Antidepressive Agents; Anxiety; Award; Behavior; Behavioral; Behavioral Assay; Binding; Biology; Brain; ChIP-seq; Chairperson; Chromatin; Chronic; Chronic stress; Data; Development; Diagnostic tests; Disease; Drug Addiction; Environment; Epigenetic Process; Exposure to; Family; Functional disorder; Gene Activation; Gene Expression; Genes; Genetic; Genetic Transcription; Genetically Engineered Mouse; Grant; HSV vector; Histone Deacetylase; Individual; Infusion procedures; Institutes; Knowledge; Lead; Life; Measures; Mediating; Mental Depression; Mentors; Mentorship; Methodology; Modeling; Molecular; Mood Disorders; Morbidity - disease rate; Mus; Neurobiology; Neurons; Neurosciences; New York; Nucleus Accumbens; Outcome; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Play; Population; Positioning Attribute; Predisposition; Prevalence; Preventive; Preventive Intervention; Property; Proteins; Regulation; Repression; Research; Research Personnel; Resveratrol; Rewards; Risk Factors; Rodent Model; Role; Scientist; Signal Pathway; Signal Transduction; Simplexvirus; Sirtuins; Stress; Technology; Testing; Therapeutic Intervention; Time; Tissue Sample; Tissues; Training; Transgenic Mice; United States National Institutes of Health; Viral Vector; Work; base; behavior influence; career; cell type; chromatin immunoprecipitation; depressive symptoms; effective therapy; experience; gene environment interaction; gene repression; genome-wide; improved; inhibitor/antagonist; interest; knock-down; medical schools; member; mortality; neuropsychiatry; next generation sequencing; novel; osmotic minipump; overexpression; programs; resilience; response; sirtinol; social; stress related disorder; therapeutic target; tool; vector

DESCRIPTION (provided by applicant): Epigenetic factors may play a key role in unraveling the molecular mechanisms underlying the pathogenesis of several neuropsychiatric disorders, including depression and drug addiction. Genes appear to explain only part of the risk factor for developing these disorders. The NIH Pathway to Independence Award (K99/R00) will significantly facilitate the candidate's ability to begin his career as an independent scientist, allowing him to study the epigenetic mechanisms underlying the gene-environment interactions observed in the onset of major mood disorders. Under the primary mentorship of Dr. Eric Nestler, Chairman of the Department of Neuroscience, and Director of the Friedman Brain Institute, at Mount Sinai School of Medicine in New York, the Pathway to Independence Awards would provide the candidate the opportunity to extend and develop his training and expertise in next generation sequencing technologies and cell type specific signaling analysis. The Award would advance the candidate's long term career objectives to: (1) determine how the environment interacts with genes and identify mechanisms by which experience confers enduring changes in gene expression, and (2) discover novel susceptibility (or resilience) genes to understand how they influence behavior. Despite the prevalence of depression and its considerable impact, knowledge about its pathophysiology is rudimentary. Thus, there is an urgent need to discover novel signaling pathways contributing to the development of depression so that better diagnostic tests, treatments, and preventive measures can be attained. The objectives of this program of research are to evaluate the role of SIRT1 and its downstream targets as potential new candidates for the treatment of neuropsychiatric disorders by performing chromatin immunoprecipitation followed by genome-wide profiling (ChIP-seq) in nucleus accumbens (NAc) tissue from control and socially defeated stressed mice. Preliminary data from our lab show that chronic social defeat stress, an ethologically validated model of depression and other stress-related disorders, modulates SIRT1 levels in the NAc and is a pro-depressant sufficient to increase stress sensitivity. The SIRT1 protein is the founding member of a family of NAD+-dependent deacetylases and ADP-ribosyltransferases, termed sirtuins. In this grant we propose to test the hypothesis that modulation of SIRT1 constitutes a novel candidate therapeutic target for antidepressants. In the mentored K-phase of this grant (Specific Aim 1 and Specific Aim 2) we will directly determine the role of sirtuins in regulating depressive- and anxiety-like behaviors using pharmacological and genetic tools. First, we will inhibit or increase the activity of sirtuins by direct intra-NAc infusion of a pharmacological inhibitior (sirtinol) or activator (resveratrol) to assess the effects of sirtuins on the development of stress-induced depressive and anxiety-like behaviors. Next, we will specifically target SIRT1 using a genetic approach to overexpress HSV-Cre viral vectors in the NAc of floxed SIRT1 (SIRT1flx) mice to knock-down SIRT1 levels, or to increase SIRT1 levels by overexpressing SIRT1 using HSV-SIRT1 vectors in the NAc. In Specific Aim 2 we also will identify transcriptional targets of SIRT1 in the NAc regulated by social defeat and antidepressants in susceptible and resilient mice by performing ChIP-Seq for SIRT1 in addition to markers of gene activation and repression. In the independent phase (R00), Specific Aim 3, will characterize the effects of social defeat on sirtuin signaling in a cell type specific manner in the NAc using drd1-GFP and drd2-GFP transgenic mice, allowing for the identification of striatonigral and striatopallidal medium spiny neurons (MSNs). In summary, the research proposed in this Pathway to Independence Award will prepare the candidate to develop a fully independent research program capable of integrating a wide range of molecular and behavioral approaches in a technically advanced and high impact manner, including: (i) lines of genetically engineered mice to target cell type specific regulation of sirtuin signaling after exposure to chronic stress, (ii) the ability to isolate chromatin from bain tissue, (iii) ChIP followed by genome-wide profiling, (iv) a rodent model of depression with high validity, and (v) automated behavioral assays measuring many depression- and anxiety-like behavioral responses. .

描述（由申请人提供）：表观遗传因素可能在阐明几种神经精神疾病（包括抑郁症和药物成瘾）发病机制的分子机制中发挥关键作用。基因似乎只能解释这些疾病的部分风险因素。NIH独立之路奖（K99/R 00）将显著促进候选人作为独立科学家开始职业生涯的能力，使他能够研究在重大情绪障碍发作中观察到的基因-环境相互作用的表观遗传机制。在纽约西奈山医学院神经科学系主任兼弗里德曼脑研究所所长Eric Nestler博士的主要指导下，独立之路奖将为候选人提供机会，以扩展和发展他在下一代测序技术和细胞类型特异性信号分析方面的培训和专业知识。该奖项将推进候选人的长期职业目标：（1）确定环境如何与基因相互作用，并确定经验赋予基因表达持久变化的机制，以及（2）发现新的易感性（或弹性）基因，以了解它们如何影响行为。尽管抑郁症的患病率及其相当大的影响，其病理生理学的知识是基本的。因此，迫切需要发现有助于抑郁症发展的新信号通路，以便获得更好的诊断测试，治疗和预防措施。这项研究计划的目的是评估SIRT 1及其下游靶点作为治疗神经精神疾病的潜在新候选物的作用，方法是在对照组和社交失败应激小鼠的神经核（NAc）组织中进行染色质免疫沉淀，然后进行全基因组分析（ChIP-seq）。我们实验室的初步数据表明，慢性社会失败压力（一种行为学验证的抑郁症和其他压力相关疾病模型）调节NAc中的SIRT 1水平，并且是一种足以增加压力敏感性的促进剂。SIRT 1蛋白是NAD+依赖性脱乙酰酶和ADP-核糖基转移酶家族的创始成员，称为sirtuins。在这项研究中，我们提出了一个假设，即SIRT 1的调节构成了抗抑郁药的一个新的候选治疗靶点。在该资助的指导K阶段（具体目标1和具体目标2），我们将使用药理学和遗传学工具直接确定sirtuins在调节抑郁和焦虑样行为中的作用。首先，我们将通过直接在NAc内输注药理学抑制剂（sirtinol）或 激活剂（白藜芦醇），以评估sirtuins对应激诱导的抑郁和焦虑样行为的发展的影响。接下来，我们将使用遗传方法特异性靶向SIRT 1，在floxed SIRT 1（SIRT 1flx）小鼠的NAc中过表达HSV-Cre病毒载体以敲低SIRT 1水平，或通过在NAc中使用HSV-SIRT 1载体过表达SIRT 1来增加SIRT 1水平。在《特定目标2》中，我们还将通过对SIRT 1进行ChIP-Seq以及基因激活和抑制的标记物，在易感和有弹性的小鼠中识别受社会失败和抗抑郁药调节的NAc中SIRT 1的转录靶点。在独立阶段（R 00），具体目标3，将使用drd 1-GFP和drd 2-GFP转基因小鼠在NAc中以细胞类型特异性方式表征社会失败对沉默调节蛋白信号传导的影响，从而鉴定纹状体黑质和纹状体中棘神经元（MSN）。总而言之，独立之路奖中提出的研究将为候选人开发一个完全独立的研究计划做好准备，该计划能够以技术先进和高影响力的方式整合广泛的分子和行为方法，包括：（i）基因工程小鼠的品系，以靶向细胞类型特异性调节 的sirtuin信号后，暴露于慢性压力，（ii）的能力，以分离染色质从贝恩组织，（iii）ChIP，其次是全基因组分析，（iv）抑郁症的啮齿动物模型具有高的有效性，和（v）自动行为测定测量许多抑郁症和焦虑样行为反应。.