TRANSCRIPTIONAL REGULATION OF ENDOCARDIAL DEVELOPMENT

心内膜发育的转录调控

基本信息

批准号：
7837505
负责人：
BIN ZHOU
金额：
$ 22.81万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-15 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7837505
关键词：
Ablation Adult Affinity Chromatography Animal Model Attention Binding Binding Sites Biological Biological Assay Cardiac Cell Death Cell Line Cell physiology Cells Complex DNA Data Defect Development Electrophoretic Mobility Shift Assay Elements Embryo Embryonic Development Endocardium Enhancers Etiology Gel Gene Expression Gene Targeting Genes Genetic Genetic Determinism Genetic Enhancer Element Genomics Growth Heart Heart Valve Diseases Heart Valves Immunoblotting Introns Knowledge Maintenance Mass Spectrum Analysis Mesenchymal Molecular Morphogenesis Mus Mutate Mutation Nucleic Acid Regulatory Sequences Pathway interactions Play Population Process Regulation Regulatory Element Research Personnel Role Site Specificity Stem cells Testing Time Tin Tissues To specify Transcriptional Regulation Transgenic Mice Ventricular septum Work base cardiogenesis congenital heart disorder critical period in vivo insight mouse model novel programs protein protein interaction spatiotemporal tissue regeneration transcription factor

项目摘要

DESCRIPTION (provided by applicant): The long-term objectives of this proposal are to understand the genetic control of a specific subpopulation of endocardium, the pro-valve endocardial cells (PVECs), and determine their role in valve formation. During embryogenesis, NFATd expression is restricted to the endocardium and its expression is accentuated in the PVECs (endocardial cells that do NOT undergo mesenchymal transformation during the formation of cardiac valves). These observations suggest that a focus on NFATd regulation may provide unique insights into the transcriptional program orchestrating pro-valve endocardial development and valve formation. Having recently identified a 243 bp sequence within the first intron of NFATd that serves as a PVEC specific transcriptional enhancer required for accentuation of NFATd gene expression and having determined that a conserved 58 bp sequence within this enhancer region functions as a critical cis-regulatory element, controlling both the intensity and specificity of NFATd expression in the PVEC population, we hypothesized that this NFATd enhancer region defines an enhanceosome that plays a critical role in the regulation of NFATd expression essential for normal PVEC development and valve formation. Therefore, we propose to 1) Determine the molecular components and interactions required for activation of the PVEC-specific NFATd transcriptional enhancing complex using a novel enodardial cell line for gel shift, DNA affinity purification and mass psectrometry, 2) Define the in vivo role of the PVEC enhanceosome in PVEC development and valve formation utilizing a cre-loxp based dual-genetic mouse model for sequential analysis of enhanceosome suppression and activation of NFATd expression, and 3) Delineate the in vivo role of the PVEC endocardial subpopulation by spatiotemporal ablation of these cells using bigenic cre-/loxp, puAtk conditional genetic ablator mouse line. This work will define critical regulatory mechanisms required for heart valve development and will inform novel tissue-regeneration remedies for valve degeneration.

描述（由申请人提供）：该提案的长期目标是了解内膜内膜，前心脏内膜细胞（PVEC）的特定亚种群的遗传控制，并确定其在瓣膜形成中的作用。在胚胎发生过程中，NFATD表达仅限于心内膜，其表达在PVEC（在心脏瓣膜形成期间不经历间充质转化的心内膜细胞）中得到了强调。这些观察结果表明，对NFATD调节的关注可能会为策划亲valve心内膜发育和瓣膜形成的转录程序提供独特的见解。最近在NFATD的第一个内含子中鉴定了243 bp序列，该序列是PVEC的特定转录增强子，以强调NFATD基因表达所需定义一个增强体，在调节正常PVEC发育和瓣膜形成必不可少的NFATD表达中起着至关重要的作用。 Therefore, we propose to 1) Determine the molecular components and interactions required for activation of the PVEC-specific NFATd transcriptional enhancing complex using a novel enodardial cell line for gel shift, DNA affinity purification and mass psectrometry, 2) Define the in vivo role of the PVEC enhanceosome in PVEC development and valve formation utilizing a cre-loxp based dual-genetic mouse model for对NFATD表达的增强抑制和激活的顺序分析，以及3）使用Begenic CRE/LOXP使用PUATK条件遗传学小鼠线来描述这些细胞的PVEC心内膜亚体系的体内作用。这项工作将定义心脏瓣膜发育所需的关键调节机制，并将为瓣膜变性提供新的组织再生疗法。