TRANSCRIPTIONAL CONTROL OF CARDIAC GROWTH

心脏生长的转录控制

• 批准号: 7609863
• 负责人: Kyu-Ho Lee
• 金额: $ 21.15万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7609863
• 关键词: Binding; Biological Assay; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cells; Cessation of life; Chromatin Remodeling Factor; Chronic Disease; Computer Retrieval of Information on Scientific Projects Database; Defect; Embryo; Funding; Future; Gene Targeting; Genes; Grant; Growth; Growth Factor; Heart; Heart Transplantation; Heart Ventricle; Human; In Vitro; Injury; Institution; Joints; Modeling; Molecular; Molecular Profiling; Muscle Cells; Mutation; Myocardium; Population; Promoter Regions; Protein Isoforms; Protein Overexpression; RNA Interference; Regulation; Research; Research Personnel; Resources; Right ventricular structure; Shapes; Signal Transduction; Source; Stem cells; Structure; System; Testing; Therapeutic; Transcriptional Regulation; United States National Institutes of Health; Ventricular; Zinc Fingers; chromatin immunoprecipitation; congenital heart disorder; design; in vivo; insight; loss of function; repaired; research study; size; stem; transcription factor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project seeks to define the molecular mechanisms controlling the growth and proliferation of heart muscle cells or cardiac myocytes in the developing embryo. Dysregulation of heart muscle proliferation in developing segments or chamber precursors of the heart leads to major forms of congenital heart disease in humans, including chamber hypoplasia and septal and valve defects causing chronic illness and early death. The research described in this proposal will investigate the interaction of two regulators of embryonic cardiac proliferation, the zinc finger domain-containing transcription factor Gata4, and its associated co-factor Smarcd3. Independent mutation or loss of function of either gene results in poor growth or hypoplasia of the right ventricle of heart due to decreased proliferation of myocyte precursors. This leads to the hypothesis that critical genes regulating right ventricle growth are subject to dual regulation by Gata4 and Smarcd3 through direct binding of Smarcd3 to Gata4, and recruitment of Smarcd3's associated SWI/SNF chromatin remodeling complex to genes binding Gata4 at their promoter regions. Experiments described in this proposal will first examine the molecular determinants of Gata4-Smarcd3 binding, both to investigate the potential for regulation by growth factor signaling of this interaction, and as a guide to designing dominant interfering isoforms of either Gata4 or Smarcd3. Overexpression of these isoforms by lentiviral transduction in the P19CL6 model cardiac myocyte system will specifically disrupt activation of Gata4-Smarcd3 joint target genes. Comparison of resulting expression profiles to those from control P19CL6 cells and cells subjected to RNA interference knockdown of Gata4 and Smarcd3 will identify a focused population of candidate genes for testing by chromatin immunoprecipitation for direct activation by Gata4 and Smarcd3. Assay of resulting candidate genes by overexpression and conditional knockdown in-vitro and in-vivo will provide the first steps in determining a molecular roadmap for right ventricular proliferation, and insight as to how the embryo regulates the size, shape and structure of its heart. Such insight will also be of great utility to future therapeutic efforts seeking to identify, culture and expand cardiac stem or progenitor cells for use in post-injury grafting and cardiac repair.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该项目旨在定义控制发育中胚胎中心肌细胞或心肌细胞生长和增殖的分子机制。 心脏发展中心或心脏室前体中心肌增殖的失调导致人类中先天性心脏病的主要形式，包括室内发育不全和瓣膜和瓣膜缺陷，导致慢性病和早期死亡。 该提案中描述的研究将研究两个胚胎心脏增殖的调节剂，含锌指域的转录因子GATA4及其相关的副因素SMARCD3的相互作用。 独立的突变或任何基因功能丧失会导致由于肌细胞前体的增殖减少而导致心脏右心室的发育不良。 这导致了以下假设：调节右心室生长的关键基因通过SMARCD3与GATA4的直接结合以及SMARCD3的SMARCD3相关的SWI/SNF染色质重塑复合物与促进者区域结合GATA4的基因与基因结合的基因的募集，对GATA4和SMARCD3进行了双重调节。 该提案中描述的实验将首先检查GATA4-SMARCD3结合的分子决定因素，以研究通过这种相互作用的生长因子信号传导调控的潜力，以及作为设计GATA4或SMARCD3的显性干扰同种型的指南。 P19CL6模型心肌细胞系统中慢病毒转导对这些同工型的过表达将特别破坏GATA4-SMARCD3关节靶基因的激活。 将所得的表达谱与受到RNA干扰敲低的对照P19CL6细胞和细胞的表达谱进行比较，将鉴定出聚焦的候选基因群，以通过染色质免疫沉淀通过GATA4和SMARCD3进行直接激活进行测试。 通过过表达和有条件敲低的体外和体体的有条件敲低的候选基因的测定将提供确定右心室增殖的分子路线图的第一步，以及有关胚胎如何调节其心脏大小，形状和结构的洞察力。 这种洞察力也将对未来的治疗努力具有很大的效用，以识别，培养和扩展心脏病或祖细胞，用于用于伤害后移植和心脏修复。