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

基本信息

批准号：
9054163
负责人：
Deveroux Ferguson
金额：
$ 23.73万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9054163
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 LoxP-flanked allele 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 Intervention Preventive measure 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 anxiety-like behavior base behavior influence behavioral response brain tissue career cell type chromatin immunoprecipitation depression model depressive symptoms effective therapy experience gene environment interaction gene repression genetic approach genome-wide improved inhibitor/antagonist interest knock-down medical schools member mortality neuropsychiatric disorder next generation sequencing novel osmotic minipump overexpression programs resilience social stress related disorder tenure track therapeutic target tool vector

项目摘要

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. .

项目概要表观遗传因素可能在揭示其背后的分子机制中发挥关键作用几种神经精神疾病的发病机制，包括抑郁症和毒瘾。基因似乎仅解释发生这些疾病的部分风险因素。 NIH 独立之路奖（K99/R00）将极大地促进候选人作为独立科学家开始其职业生涯的能力，使他能够研究在观察到的基因与环境相互作用背后的表观遗传机制。严重情绪障碍的发作。在系主任 Eric Nestler 博士的指导下纽约西奈山医学院神经科学博士兼弗里德曼大脑研究所所长独立之路奖将为候选人提供扩展和发展自己的机会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。这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赋予基因持久变化的机制表达，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。尽管抑郁症很普遍且影响相当大，但对其的了解病理生理学是初级的。因此，迫切需要发现新的信号通路有助于抑郁症的发展，以便更好的诊断测试、治疗和预防措施可以达到。该研究计划的目标是评估 SIRT1 的作用及其下游目标作为治疗神经精神疾病的潜在新候选者伏隔核 (NAc) 中染色质免疫沉淀随后进行全基因组分析 (ChIP-seq) 来自对照小鼠和在社会上失败的应激小鼠的组织。我们实验室的初步数据表明，慢性社交战胜压力是一种经过行为学验证的抑郁症和其他压力相关疾病模型，可以调节 SIRT1 水平存在于 NAc 中，是一种足以提高应激敏感性的促抑郁剂。 SIRT1 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，称为瑟土因。在这项资助中，我们建议测试 SIRT1 调制构成新候选者的假设抗抑郁药的治疗目标。在本次资助的指导 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学和遗传工具。首先，我们将通过直接intra-NAc来抑制或增加sirtuins的活性输注药理抑制剂（西替诺）或激活剂（白藜芦醇）以评估去乙酰化酶对压力诱发的抑郁和焦虑样行为的发展。接下来，我们将专门针对SIRT1 使用遗传方法在 floxed SIRT1 (SIRT1flx) 小鼠的 NAc 中过度表达 HSV-Cre 病毒载体敲低 SIRT1 水平，或通过使用 HSV-SIRT1 载体过表达 SIRT1 来增加 SIRT1 水平 NAc。在具体目标 2 中，我们还将确定 NAc 中受社会调节的 SIRT1 转录靶点通过对 SIRT1 进行 ChIP-Seq，除了基因激活和抑制的标记。在独立阶段 (R00)，具体目标 3 将表征使用 drd1-GFP 在 NAc 中以细胞类型特异性方式社交失败对 Sirtuin 信号传导的影响 drd2-GFP 转基因小鼠，可识别纹状体黑质和纹状体苍白质中等刺状神经元（MSN）。总之，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，能够整合广泛的分子以技术先进和高影响力的方式进行行为方法，包括：（i）遗传系工程小鼠在暴露于慢性应激后能够针对细胞类型特异性调节沉默调节蛋白信号传导，（ii）从脑组织中分离染色质的能力，(iii) ChIP，然后进行全基因组分析，(iv) 啮齿动物具有高有效性的抑郁症模型，以及（v）测量许多抑郁症和抑郁症的自动行为测定类似焦虑的行为反应。。