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: 8830503
• 负责人: Deveroux Ferguson
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8830503
• 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; brain tissue; 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

Project Summary 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 brain 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. .

项目摘要 表观遗传因素可能在解开其背后的分子机制方面发挥关键作用。 几种神经精神障碍的发病机制，包括抑郁症和药物成瘾。基因似乎可以 只解释导致这些疾病的风险因素的一部分。美国国立卫生研究院独立之路奖 (K99/R00)将大大促进候选人作为独立科学家开始其职业生涯的能力， 使他能够研究基因-环境相互作用背后的表观遗传机制 出现严重的情绪障碍。在该部门主席埃里克·内斯特勒博士的主要指导下 他是纽约西奈山医学院弗里德曼脑研究所的主任， 独立之路奖将为候选人提供扩展和发展他的机会 在下一代测序技术和特定细胞类型信号分析方面的培训和专业知识。这个 奖励将推动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持久变化的机制 表达，以及(2)发现新的易感(或弹性)基因以了解它们如何影响 行为。尽管抑郁症的流行及其相当大的影响，但对其 病理生理学是最基本的。因此，迫切需要发现新的信号通路 有助于抑郁症的发展，以便更好地进行诊断测试、治疗和预防 这些措施是可以实现的。这项研究的目标是评估SIRT1及其 下游靶点作为治疗神经精神障碍的潜在新候选药物 染色质免疫沉淀和全基因组图谱(ChIP-SEQ)在伏核(NAC)中的应用 来自对照组和被社会打败的应激小鼠的组织。我们实验室的初步数据显示，慢性社交 击败压力，一种经行为学验证的抑郁症和其他与压力相关的障碍的模型，调节 NAC中的SIRT1水平，是一种足以增加应激敏感性的促抑郁药。SIRT1蛋白是 依赖NAD+的脱乙酰酶和ADP-核糖基转移酶家族的创始成员，称为 先生们。在这项授权中，我们建议检验SIRT1的调制构成一个新的候选者的假设 抗抑郁药物的治疗靶点。在这笔赠款的指导K阶段(具体目标1和具体目标 2)我们将直接确定sirtuins在调节抑郁和焦虑样行为中的作用 药理和遗传工具。首先，我们将通过直接NAC抑制或增强sirtuins的活性 输注药物抑制剂(Sirtinol)或激活剂(白藜芦醇)以评估sirtuins对 压力导致的抑郁和焦虑样行为的发展。接下来，我们将专门针对SIRT1 用遗传学方法在SIRT1(SIRT1flx)小鼠NAC中高表达HSV-Cre病毒载体 下调SIRT1水平，或通过使用HSV-SIRT1载体过表达SIRT1来增加SIRT1水平 南华早报。在特定目标2中，我们还将确定SIRT1在受社会调控的NAC中的转录靶点 在易感和有弹性的小鼠中通过对SIRT1执行芯片序列来失败和抗抑郁药物 基因激活和抑制的标志物。在独立阶段(R00)，具体目标3将表征 应用DRD1-GFP和NAC研究社会失败对NAC细胞类型特异性sirtuin信号转导的影响 DRD2-GFP转基因小鼠，可用于纹状体黑质和纹状体苍白质中间刺的鉴定 神经元(MSN)。总而言之，这一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，能够整合广泛的分子 以技术先进和影响大的方式采取的行为方法，包括：(1)遗传品系 基因工程小鼠在慢性应激后靶向细胞类型特异性调节sirtuin信号，(Ii) 从脑组织中分离染色质的能力，(Iii)芯片和全基因组图谱，(Iv)啮齿动物 具有高有效性的抑郁症模型，以及(V)测量许多抑郁症的自动行为分析-以及 焦虑样的行为反应。 。