Neuronal subtype and circuit-specific epigenetic mechanisms in addiction

成瘾中的神经元亚型和回路特异性表观遗传机制

基本信息

批准号：
9260817
负责人：
Gregory Charles Sartor
金额：
$ 13.1万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9260817
关键词：
Alpha Cell Anatomy Animal Model Area Attenuated BRD2 gene Behavior Binding Proteins Brain region Bromodomain Canine Adenoviruses Chromatin Chronic Clinical Trials Cocaine Cocaine Dependence Complex Corpus striatum structure Data Development Dissection Dopamine Drug Addiction Emotional Enterobacteria phage P1 Cre recombinase Epigenetic Process Faculty Family Fluorescence-Activated Cell Sorting Functional disorder Gene Expression Gene Transfer Gene Transfer Techniques Genes Genetic Transcription Goals Grant Gray unit of radiation dose Histone Acetylation Histones Individual Injectable Journals Lysine Macromolecular Complexes Mediating Mentors Messenger RNA Methods Modeling Modification Molecular Molecular Biology Techniques Molecular Genetics Neurons Nucleus Accumbens Pathway interactions Pharmacology Phase Play Population Positioning Attribute Presynaptic Terminals Promoter Regions Proteins Publishing Rattus Reader Records Recruitment Activity Request for Proposals Research Rewards Role Scientific Advances and Accomplishments Self Administration Societies Supervision Technical Expertise Technology Time Tracer Training United States National Institutes of Health Viral addiction behavioral pharmacology behavioral response career career development cell type chromatin modification craving drug seeking behavior economic cost effective therapy epigenetic drug epigenetic regulation global health improved innovation knock-down member nano-string neural circuit neuroadaptation neuronal cell body novel preference protein complex protein expression public health relevance receptor relating to nervous system retrograde transport scaffold skills small hairpin RNA tool

项目摘要

DESCRIPTION (provided by applicant): This proposal requests support for a comprehensive training plan that will enable the candidate, Dr. Gregory C. Sartor, to expand, develop, and refine technical skills that are necessary for a productive independent research career. The goal of the training plan is to acquire advanced molecular biology techniques through supervised hands-on training and didactic coursework in order to incorporate sophisticated methods into Specific Aims focused on elucidating cell type- and circuit-specific epigenetic mechanisms in addiction. The advanced scientific training will be focused on two key areas: 1) Fluorescence-activated cell sorting (FACS) of neuronal subtypes from addiction-related brain regions, and 2) developing viral-mediated gene transfer tools to manipulate epigenetic targets in a cell type- and connection-specific fashion. The career development activities will be mentored by Dr. Claes Wahlestedt with collaborative support from Drs. Kasahara and Gray, all of which have non-overlapping expertise in the methods proposed and have exceptional records of mentoring postdoctoral trainees. The research component will investigate acetyl lysine reader proteins, termed BET bromodomains, an emerging, drugable epigenetic target that is currently being studied in multiple clinical trials. These bromodomain-containing proteins bind acetylated histones and serve as a scaffold for the recruitment of macromolecular complexes that modify chromatin accessibility and transcriptional activity. I recently found that BET proteins are upregulated and recruited to the promoter region of Bdnf in the nucleus accumbens (NAc) following chronic cocaine administration. Furthermore, my data show that pharmacological inhibition of these BET proteins (systemic and intra-NAc) attenuates behavioral responses to cocaine. However, recent evidence indicates that cocaine-induced behaviors and associated chromatin modifications are regulated in a cell type-specific manner in the NAc. Therefore, precise examinations of BET proteins within this anatomically and functionally heterogeneous region are critically needed to fully understand the role of these proteins in complex addiction behaviors. My overall hypothesis is that modulation of epigenetic reader proteins in a cell type- and projection-dependent manner will attenuate cocaine seeking behaviors. Aim 1 will require training in FACS in order to determine the cell-type specific time course of BET mRNA/protein expression following short and long access to cocaine self-administration. Aim 2 will require proficiency in viral-mediated gene transfer techniques in order to manipulate BET proteins in specific cell types within the NAc during cocaine seeking behaviors. The R00 independent phase builds on this training to focus more specifically on connection-specific changes and manipulations of BET proteins during cocaine seeking behaviors. Together, these innovative studies will be the first to systemically explore epigenetic targets in cell type- and connection- specific manner during complex behaviors.

描述（由适用提供）：该提案要求为一项全面的培训计划提供支持，该计划将使候选人Gregory C. Sartor博士扩大，开发和完善产品独立研究职业所必需的技术技能。培训计划的目的是通过监督的动手培训和教学课程获得先进的分子生物学技术，以便将复杂的方法纳入成瘾中阐明细胞类型和电路特异性表观遗传机制的特定目标。先进的科学训练将集中在两个关键领域：1）与成瘾相关的大脑区域的神经元亚型的荧光激活细胞分选（FACS），以及2）开发病毒介导的基因转移工具，以操纵细胞类型和连接特定的特定方式中的表观遗传靶标。 Claes Wahlestedt博士将在Drs的合作支持下考虑职业发展活动。卡萨哈拉（Kasahara）和格雷（Gray），所有这些都在提出的方法方面具有不重叠的专业知识，并且具有精神上的博士后学员记录。该研究成分将研究乙酰赖氨酸读蛋白蛋白，称为BET BOTORODOMARIES，这是一种新兴的，可吸毒的表观遗传靶标，目前正在多个临床试验中研究。这些含溴脱域的蛋白结合了乙酰化组蛋白，并用作募集大分子复合物的支架，从而改变了染色质的可及性和转录活性。我最近发现，在慢性可卡因给药后，BET蛋白被上调并募集到原子核（NAC）中BDNF的启动子区域。此外，我的数据表明，对这些BET蛋白（全身性和NAC内）的药理抑制作用减弱了对可卡因的行为反应。但是，最近的证据表明，可卡因诱导的行为和相关的染色质修饰以NAC的细胞类型特异性调节。因此，在这种解剖学和功能上异质区域内对BET蛋白的精确检查至关重要，以充分了解这些蛋白质在复杂的添加行为中的作用。我的总体假设是，以细胞类型和投影依赖性方式对表观遗传读取器蛋白的调节将减弱可卡因的寻求行为。 AIM 1将需要在FACS中进行训练，以确定在短而长的可卡因自我管理后，BET mRNA/蛋白质表达的细胞类型特定时间过程。 AIM 2将需要熟悉病毒介导的基因转移技术，以便在可卡因寻求行为的情况下操纵NAC内特定细胞类型的BET蛋白。 R00独立阶段建立在此培训的基础上，更具体地说，在可卡因寻求行为期间，针对特定连接的变化和操纵。总之，这些创新的研究将是第一个在复杂行为中系统地探索细胞类型和连接方式的表观遗传靶标的。