Coup-tf dependent mechanisms of ventricular and hemangioblast specification

心室和成血管细胞规范的 Coup-tf 依赖性机制

• 批准号: 8606886
• 负责人: Joshua Waxman
• 金额: $ 33.74万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-15 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8606886
• 关键词: Adult; Affect; Biological Assay; Blood Cells; Blood Vessels; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell Lineage; Cell Transplantation; Cell physiology; Cells; Chickens; Child; Complement; Congenital Abnormality; Coupled; Data; Defect; Development; EMSA; Endothelial Cells; Event; FGF8 gene; Family; Fibroblast Growth Factor; Future; Genes; Genetic; Genetic Epistasis; Glean; Goals; Grant; Healed; Hematopoietic; Homologous Gene; Human; In Situ Hybridization; In Vitro; Invertebrates; Lateral; Light; Luciferases; Maps; Mesoderm; Methods; Molecular; Molecular Genetics; Mus; Mutation; Nature; Nuclear Hormone Receptors; Orphan; Ovalbumin; Population; Prevalence; Process; Research; Role; Signal Transduction; Stem cells; Testing; Tissues; Transcription Coactivator; Transcription Repressor/Corepressor; Tretinoin; Ventricular; Work; Zebrafish; base; cardiogenesis; chromatin immunoprecipitation; healing; heart dimension/size; improved; in vivo; injured; loss of function; malformation; novel; prevent; progenitor; promoter; public health relevance; regenerative therapy; repaired; research study; stem cell therapy; transcription factor

DESCRIPTION (provided by applicant): Congenital cardiovascular malformations are among the most common congenital birth defects, occurring in almost 1% of the population. However, we do not understand the underlying molecular nature of the majority of these defects. In order to develop effective in vitro stem cell and regenerative therapies aimed at preventing birth defects and healing cardiovascular diseases in adults, it is critical to have a precise understanding of the mechanisms directing cardiovascular development in vivo. Therefore, the long-term goal of our lab is to understand the mechanisms of cardiovascular specification during vertebrate development. The specific aims of this grant are to elucidate the mechanisms by which chicken ovalbumin upstream promoting-transcription factors (Coup-tfs) are required to restrict ventricular specification and promote hemangioblast specification using zebrafish. In humans, Coup-tfs are required for normal heart development. Studies in mice have demonstrated that Coup-tf2 is required for proper early cardiovascular development, but the mechanisms underlying Coup-tf2 function in these early defects are not understood. Our preliminary results suggest that zebrafish Coup-tf1a has cardiovascular defects reminiscent of mammalian Coup-tf2. Moreover, our preliminary results support the hypothesis that zebrafish Coup-tf1a has distinct requirements restricting ventricular cell specification and promoting hemangioblast specification. In Specific Aim 1, we will use cell transplantation and lineage tracing experiments to determine which cells required Coup-tf1a to restrict ventricular cell specification. We will also determine the genetic relationship of FGF8 and Coup-tf1a in restricting ventricular cell specification. A role for Coup-tfs in vertebrate hemangioblast specification has not been previously recognized. In Specific Aim 2, we will use epistasis and cell transplantation analysis to determine the relationship of Coup-tf1a to known regulators of hemangioblast specification and what cells require Coup-tf1a for hemangioblast specification. Factors downstream of Coup-tf1a in ventricular cell and hemangioblast specification are not known. In Specific Aim 3, we will use loss-of-function methods to determine if candidate genes are required for regulating these distinct processes downstream of Coup-tf1a. Altogether, these studies will improve our understanding of normal cardiovascular specification events during development, providing the basis for future therapies aimed at healing congenital cardiovascular defects in children and injured or diseased cardiovascular tissues in adults.

描述（由申请人提供）：先天性心血管畸形是最常见的先天性先天出生缺陷，几乎是1％的人口。但是，我们不了解大多数这些缺陷的基本分子性质。为了开发有效的体外干细胞和旨在预防成人先天缺陷和愈合心血管疾病的再生疗法，至关重要的是，要精确了解导致体内心血管发育的机制。因此，我们实验室的长期目标是了解脊椎动物发育过程中心血管规范的机制。该赠款的具体目的是阐明需要促进鸡卵形蛋白上游促进转录因子（COUP-TFS）的机制，以限制心室规格并使用斑马鱼促进血管母细胞规范。在人类中，正常心脏发展需要政变。在小鼠中的研究表明，coup-tf2是适当的早期心血管发育所必需的，但是在这些早期缺陷中的coup-tf2功能的机制尚不清楚。我们的初步结果表明，斑马鱼政变 - TF1A具有心血管缺陷，让人联想到哺乳动物COUP-TF2。此外，我们的初步结果支持以下假设：斑马鱼政变TF1A具有限制心室细胞规范和促进血管细胞规范的不同要求。在特定的目标1中，我们将使用细胞移植和谱系跟踪实验来确定哪些细胞需要COUP-TF1A限制心室细胞规范。我们还将在限制心室细胞规范中确定FGF8和COUP-TF1A的遗传关系。以前尚未认识到政变TF在脊椎动物血管细胞规范中的作用。在特定的目标2中，我们将使用上毒和细胞移植分析来确定政变-TF1A与已知的血管细胞规范调节剂的关系，以及哪些细胞需要coup-tf1a用于血管细胞规格。尚不清楚Coup-TF1A在心室细胞和血管母细胞规范中的下游因素。在特定目标3中，我们将使用功能丧失方法来确定是否需要候选基因来调节政变-TF1A下游的这些不同过程。总的来说，这些研究将提高我们对发育过程中正常心血管规范事件的理解，为未来的疗法提供了旨在治愈成人儿童以及受伤或患病的心血管组织的未来疗法的基础。