Genetic regulation of atrial gene expression in development and disease

发育和疾病中心房基因表达的遗传调控

基本信息

批准号：
10576399
负责人：
William Tswenching Pu
金额：
$ 60.73万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10576399
关键词：
Adult Affect Affinity Age Years Arrhythmia Atrial Fibrillation Binding Biological Assay Biotinylation Cardiac Cardiac Myocytes Cardiovascular system Cells ChIP-seq Chromatin Data Dependovirus Development Disease Dissection Dose EP300 gene Enhancers Etiology GATA4 gene Gene Expression Gene Expression Regulation Gene Mutation Gene Silencing Gene Transduction Agent Genes Genetic Genetic Transcription Genetic study Goals Heart Heart Atrium Homeostasis Human Genetics Individual Knock-out Knowledge Link Machine Learning Maps Measurement Measures Mechanical Stress Metabolic stress Molecular Morphology Mus Muscle Cells Mutagenesis Mutation Nodal Pathogenesis Persons Phenotype Physiological Population Precipitation Predisposition Prevalence Reagent Regulation Regulatory Element Reporter Reproducibility Research Proposals Risk Signal Transduction Techniques Testing Therapeutic Transcription Coactivator Variant Ventricular Work biochip combinatorial dosage experimental study functional genomics gene regulatory network gene therapy genome wide association study genomic locus heart function in vivo insight loss of function mosaic muscle enhancer factor-2A novel novel strategies organ growth prevent programs stem tool transcription factor

项目摘要

SUMMARY Precise regulation of atrial gene expression is crucial to maintain atrial homeostasis, and disorders or gene mutations that impact atrial gene expression can cause atrial fibrillation (AF), a serious arrhythmia that affects an estimated 33 million people worldwide. However, there remain many gaps in our understanding of atrial gene regulation, including the mechanisms that underlie chamber-selective gene expression. Human genetic studies have shown that sequence variants near the cardiac transcription factor gene TBX5 are associated with greater AF risk, and mice with Tbx5 deficiency develop AF. Although recent work has identified some TBX5-regulated genes that contribute to AF susceptibility, the atrial TBX5-centered transcriptional network is incompletely explored. Elucidation of this network and its sensitivity to TBX5 dose would reveal nodal points in AF pathogenesis and may suggest new approaches to prevent or treat AF. The overarching goal of this re- search proposal is to elucidate the atrial gene regulatory network and how it is perturbed in AF. The proposal buillds on novel reagents and techniques developed in the Pu lab to interrogate transcriptional mechanisms in vivo, including highly sensitive and reproducible cardiac transcription factor ChIP-seq through in vivo biotinyla- tion (bioChIP-seq), massively parallel in vivo measurement of cis-regulatory element (CRE) activity (AAV- MPRA assay), and mosaic gene inactivation strategies to hone in direct, cell autonomous effects of gene inac- tivation. In Aim 1, we use bioChIP-seq and AAV-MPRA to define atrial CREs and to dissect the sequence fea- tures required for their chamber selective activity. In Aim 2, we determine the effect of TBX5 deficiency on the occupancy of other TFs and p300, the activity of CREs, and the expression of atrial genes. We use these data to define the TBX5-centered atrial gene regulatory network, and to determine how this network is perturbed by TBX5 haploinsufficiency or knockout. In Aim 3, we test the hypothesis, suggested by our preliminary data, that TBX5 regulates atrial genes through functional and physical interaction with TEAD1. Successful completion of this proposal will lead to new insights into atrial gene regulation and its perturbation in AF.

概括心房基因表达的精确调节对于维持心房稳态和疾病或基因至关重要影响心房基因表达的突变会导致房颤（AF），这是一种严重的心律不齐，影响全球估计有3300万人。但是，我们对房屋的理解仍然存在很多差距基因调节，包括基于腔室选择性基因表达的机制。人遗传研究表明，心脏转录因子附近的序列变体基因TBX5与具有更大的AF风险，并且具有TBX5缺乏症的小鼠会出现AF。尽管最近的工作已经确定了一些 TBX5调节的基因有助于AF易感性，心房以TBX5为中心的转录网络是不完全探索。阐明该网络及其对TBX5剂量的敏感性将揭示淋巴结 AF发病机理，可能提出预防或治疗AF的新方法。这个重新的总体目标搜索建议是为了阐明心房基因调节网络及其在AF中的干扰。提案在PU实验室开发的新试剂和技术上的构造，以询问转录机制体内通过体内生物素 - 包括高度敏感和可重复的心脏转录因子chip-seq Tion（Biochip-Seq），在体内测量顺式调节元件（CRE）活性（AAV-- MPRA分析）和镶嵌基因灭活策略，以磨练基因的直接细胞自主作用 tivation。在AIM 1中，我们使用Biochip-Seq和Aav-Mpra来定义房屋cr板并剖析序列fea- 其室选择活动所需的调查。在AIM 2中，我们确定了TBX5缺乏对其他TF和P300的占用率，CRE的活性以及心房基因的表达。我们使用这些数据定义以TBX5为中心的心房基因调节网络，并确定该网络如何受到扰动 TBX5单倍薄或敲除。在AIM 3中，我们测试了我们的初步数据提出的假设，即 TBX5通过与TEAD1的功能和物理相互作用来调节心房基因。成功完成该提案将导致对房屋调节及其在AF中的扰动的新见解。