Developmental regulation of cardiac gene expression

描述（由申请人提供）： 该申请的目的是进一步发展候选人在分子和发育生物学方面的技能，以获得作为心血管疾病和发育研究者的独立地位。 候选人目前是心胸外科助理教授，具有心脏分子生物学基础。 这项工作将在强有力的机构支持和包括心血管生物学强有力的基础研究计划的环境下进行。 该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，由肌肉分化分子和发育生物学领域的领导者和专家Charles Ordahl教授担任导师。 该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，这些因子在发育过程中控制心脏特异性基因表达的调节。 由于统一的机制可能支配分化过程中心脏基因的协调表达，我们的假设是，心脏表型的规范需要“主”调控转录因子的活性。 禽类心肌肌钙蛋白T启动子将用作模型基因。 本申请中提出的研究涉及（1）启动子序列的突变分析，认为这对心脏特异性基因表达至关重要（2）DNA-蛋白结合研究，蛋白水解作图和克隆，以帮助识别激活心脏启动子的已知和新型转录因子，以及（3）体外和体内转染单个或多个心脏转录因子以激活心脏特异性基因启动子。 转录增强因子-1，一个多基因家族，我们最近克隆和表征，在心脏基因调控的作用将使用分子和经典胚胎学技术相结合的研究。 心脏决定因素的鉴定可能被利用为生物工程心脏成肌细胞的细胞移植在心力衰竭由于后天性和先天性心脏病的发展。 对控制基因表达的转录机制的理解 心脏发生也可以阐明各种先天性心脏缺陷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.