Developmental regulation of cardiac gene expression

描述(由申请人提供)： 这项申请的目的是为了进一步发展候选人在分子和发育生物学方面的技能，以获得心血管疾病和发育方面的独立研究人员的地位。候选人目前是心胸外科的助理教授，具有心脏分子生物学的基本基础。这项工作将在强有力的机构支持和包括强大的心血管生物学基础研究计划的环境下进行。这项研究计划将在加州大学旧金山分校心血管研究所的社区内进行，在肌肉分化的分子和发育生物学领域的领导者和专家查尔斯·奥尔达尔教授的指导下进行。 这个项目的目的是研究心脏启动子基序和核转录因子的组合相互作用，这些转录因子在发育过程中调控心脏特异基因的表达。由于统一机制可能控制着心脏基因在分化过程中的协调表达，因此我们的假设是，心脏表型的指定需要“主要”调控转录因子的活性。将以禽类心肌肌钙蛋白T启动子作为模型基因。这项应用中拟议的研究涉及(1)被认为对心脏特异基因表达至关重要的启动子序列的突变分析(2)DNA-蛋白质结合研究、蛋白质分解定位和克隆以帮助识别激活心脏启动子的已知和新的转录因子，以及(3)体外和体内单个或多个心脏转录因子的转染以激活心脏特异基因启动子。转录增强因子-1是我们最近克隆和鉴定的一个多基因家族，我们将结合分子和经典胚胎学技术来研究它在心脏基因调控中的作用。心脏决定因素的鉴定可能被用于开发生物工程心肌成肌细胞，用于细胞移植治疗获得性心脏病和先天性心脏病所致的心力衰竭。对控制基因转录机制的理解 心脏发生学也可能阐明各种先天性心脏病和心肌病的分子发病机制。

DESCRIPTION (provided by applicant): The purpose of this application is to further develop the candidate's skills in molecular and developmental biology towards attaining independent status as an investigator in cardiovascular disease and development. The candidate is currently an Assistant Professor in Cardiothoracic Surgery with a basic foundation in cardiac molecular biology. The work will be performed with strong institutional support and an environment that includes a strong basic research program in cardiovascular biology. The research plan will be performed within the community of the cardiovascular research institute of UCSF, under the mentors hip of Professor Charles Ordahl, a leader and expert in molecular and developmental biology of muscle differentiation. The aims of this project are to study the combinatorial interactions of cardiac promoter motifs and nuclear transcription factors that govern the regulation of cardiac-specific gene expression during development. Since unifying mechanisms probably govern the coordinate expression of cardiac genes during differentiation it is our hypothesis that specification of the cardiac phenotype requires the activity of "master" regulatory transcription factors. The avian cardiac troponin T promoter will be used as a model gene. The proposed research in this application involves (1) mutational analysis of promoter sequences believed to be essential for cardiac-specific gene expression (2) DNA-protein binding studies, proteolytic mapping and cloning to help identify known and novel transcription factors that activate cardiac promoters, and (3) transfection of single or multiple cardiac transcription factors in vitro and in vivo to activate cardiac-specific gene promoters. The role of transcriptional enhancer factor-1, a multigene family that we have recently cloned and characterized, in cardiac gene regulation will be investigated using a combination of molecular and classical embryological techniques. The identification of cardiac determination factors may be exploited for the development of bioengineered cardiac myoblasts for cellular transplantation in heart failure due to acquired and congenital heart disease. An understanding of the transcriptional mechanisms that control cardiogenesis may also elucidate the molecular pathogenesis of various congenital heart defects and cardiomyopathies.