CARDIAC TROPONIN T MUTATIONS AND HEART FUNCTION

心肌肌钙蛋白 T 突变与心脏功能

基本信息

批准号：
6615661
负责人：
AMY J SEHNERT
金额：
$ 12.18万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-08-04 至 2005-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6615661
关键词：
developmental genetics gene mutation genetic manipulation genetically modified animals heart contraction heart function molecular pathology protein structure function troponin vertebrate embryology zebrafish

项目摘要

The broad objective of this application is to study the molecular basis of cardiac contractility using zebrafish, a model vertebrate system. The proposed research will be conducted within the sound scientific environment at the University of California, San Francisco (UCSF). Dr. Sehnert, a fellow in Pediatric Cardiology at UCSF, has dedicated the past two years to full-time research in the sponsor's laboratory as a fellow of the Pediatric Scientist Development Program (PSDP). She is currently funded by the PSDP for a third year in the laboratory, and her immediate goals are to bring her preliminary work to publication. The candidate's long-term career goals are to establish an investigative research career studying congenital and genetically acquired heart disease. The career development plan incorporated in this application includes mentorship by a scientific sponsor and advisory committee; participation in relevant course work, seminars, and scientific retreats both locally and nationally; and an academic appointment providing protected research time to ensure her success in achieving her career goals. The proposed research plan specifically addresses the affect of cardiac troponin T (cTnT) mutations on cardiac contractility. cTnT is a crucial thin filament protein of the contractile apparatus, or sarcomere. It is clinically relevant to the human genetic disorder, familial hypertrophic cardiomyopathy (FHC). 15 percent of FHC cases result from cTnT mutations, which for unknown reasons predispose patients to sudden death despite minimally detectable hypertrophy. We hypothesize that a genetic mutation affecting cTnT expression results in the non-contractile heart defect observed in zebrafish silent heart mutant embryos. Furthermore, we believe that the visual transparency and genetic manipulability of zebrafish embryos offer distinct advantages for studying the affect of induced cTnT mutations on heart function. Therefore, we propose the following specific aims: 1) To isolate and characterize the silent heart gene in zebrafish using genetic techniques, 2) To rescue the silent heart phenotype by replacing the wild-type gene in mutant embryos, and 3) To generate transgenic zebrafish for combined in vivo and in vitro evaluation of mutant cTnT expression on heart function. Accomplishing these will advance our understanding of the regulation and in vivo role of cTnT in sarcomere assembly and cardiac contractility and will offer insight into the molecular mechanisms leading from cTnT mutation to disease.

该应用的广泛目的是使用模型脊椎动物系统斑马鱼研究心脏收缩的分子基础。拟议的研究将在加利福尼亚大学旧金山大学（UCSF）的声音科学环境中进行。 UCSF的儿科心脏病学研究员Sehnert博士将过去两年致力于在赞助商实验室担任儿科科学家发展计划（PSDP）的专家研究。她目前由PSDP资助在实验室第三年，她的直接目标是将她的初步工作发表。候选人的长期职业目标是建立研究性研究职业，研究先天性和遗传性心脏病。本申请中纳入的职业发展计划包括科学赞助商和咨询委员会的指导；参与相关课程，研讨会和科学务虚会，在当地和全国范围内；以及一个学术任命，提供受保护的研究时间，以确保她成功实现自己的职业目标。拟议的研究计划专门涉及心脏肌钙蛋白T（CTNT）突变对心脏收缩性的影响。 CTNT是收缩仪或肌膜的至关重要的薄丝蛋白。它在临床上与人类遗传疾病，家族性心肌病（FHC）相关。 FHC病例中有15％是由CTNT突变引起的，由于未知的原因，尽管肥大最少，但由于未知的原因，患者仍会猝死。我们假设影响CTNT表达的基因突变会导致斑马鱼沉默心脏突变体胚胎中观察到的非收缩心脏缺陷。此外，我们认为斑马鱼胚胎的视觉透明度和遗传可操作性为研究诱导的CTNT突变对心脏功能的影响提供了明显的优势。 Therefore, we propose the following specific aims: 1) To isolate and characterize the silent heart gene in zebrafish using genetic techniques, 2) To rescue the silent heart phenotype by replacing the wild-type gene in mutant embryos, and 3) To generate transgenic zebrafish for combined in vivo and in vitro evaluation of mutant cTnT expression on heart function. 完成这些将促进我们对CTNT在肌膜组装和心脏收缩性中的调节和体内作用的理解，并将深入了解从CTNT突变到疾病的分子机制。