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: 8299355
• 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/8299355
• 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; 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. . PUBLIC HEALTH RELEVANCE: The objective of this project is to prepare Dr. Deveroux Ferguson to transition to a position as an independent, tenure-track academic investigator. This project focuses on the role of the histone deacetylase, SIRT1, in depression models and in antidepressant action. The improved knowledge of depression and antidepressant mechanisms derived from this project promises to lead to the development of more effective treatments.

描述(由申请人提供)：表观遗传因素可能在解开几种神经精神疾病发病机制的分子机制中发挥关键作用，包括抑郁症和药物成瘾。基因似乎只能解释导致这些疾病的风险因素的一部分。NIH独立之路奖(K99/R00)将极大地促进候选人作为一名独立科学家开始其职业生涯的能力，使他能够研究在重大情绪障碍发生时观察到的基因-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扩展和发展其在下一代测序技术和细胞类型特定信号分析方面的培训和专业知识的机会。该奖项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，并确定经验导致基因表达持久变化的机制，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影响相当大，但关于其病理生理学的知识仍然很少。因此，迫切需要发现有助于抑郁症发展的新的信号通路，以便能够获得更好的诊断测试、治疗和预防措施。本研究的目的是评估SIRT1及其下游靶基因作为潜在的神经精神疾病治疗新候选基因的作用，方法是在对照组和社会挫败应激小鼠的伏隔核(NAC)组织中进行染色质免疫沉淀和全基因组图谱(CHIP-SEQ)。我们实验室的初步数据显示，慢性社会失败应激是抑郁症和其他应激相关障碍的行为学验证模型，它调节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sirtuins。在这项授权中，我们建议检验SIRT1的调节构成抗抑郁药物的新候选治疗靶点的假设。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在使用药理学和遗传工具调节抑郁和焦虑样行为方面的作用。首先，我们将抑制或增加sirtuins的活性，方法是在NAC内直接输注药物抑制剂(Sirtinol)或 激活剂(白藜芦醇)，以评估sirtuins对压力诱导的抑郁和焦虑样行为发展的影响。接下来，我们将使用一种遗传方法来针对SIRT1，在SIRT1(SIRT1flx)小鼠的NAC中过表达HSV-Cre病毒载体，以降低SIRT1水平，或者通过在NAC中使用HSV-SIRT1载体过表达SIRT1来提高SIRT1水平。在特定的目标2中，我们还将通过对SIRT1进行CHIP-SEQ以及基因激活和抑制的标记，在易感和有弹性的小鼠中通过对SIRT1进行CHIP-SEQ来确定受社会失败和抗抑郁药物调控的NAC中SIRT1的转录靶点。在独立阶段(R00)，特定目标3将在NAC中以一种细胞类型特定的方式表征社会失败对sirtuin信号的影响，使用DRD1-GFP和DRD2-GFP转基因小鼠，允许鉴定纹状体黑质和纹状体苍白质中等刺神经元(MSN)。综上所述，本独立途径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，该研究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)针对细胞类型特定调控的基因工程小鼠品系 (I)从贝恩组织分离染色质的能力；(Iii)芯片和全基因组图谱；(Iv)高效度的抑郁症啮齿动物模型；以及(V)测量许多抑郁和焦虑样行为反应的自动化行为分析。。 与公共卫生相关：这个项目的目标是让德弗鲁·弗格森博士做好准备，成为一名独立的终身教职学术研究员。本项目的重点是组蛋白脱乙酰酶SIRT1在抑郁症模型和抗抑郁作用中的作用。这个项目对抑郁症和抗抑郁剂机制的了解有所改善，有望导致更有效的治疗方法的开发。