RAF KINASE AND EPIGENETIC REGULATION OF FETAL VASCULAR DEVELOPMENT

RAF 激酶和胎儿血管发育的表观遗传调控

• 批准号: 9134923
• 负责人: Ravi Goyal
• 金额: $ 39.5万
• 依托单位: LOMA LINDA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-11 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9134923
• 关键词: Adipocytes; Adult; Apoptosis; Apoptotic; Arteries; BRAF gene; Basic Science; Binding; Biological Assay; Blood Vessels; CD34 gene; Cell Death; Cell Survival; Cells; Cerebral Palsy; Clinical; Collagen; DNA; DNA Methylation; Data; Data Reporting; Defect; Development; Dioxygenases; Disease; Embryo; Endothelial Cells; Epigenetic Process; Family; Fetal Development; Fibroblasts; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Growth; Health; Hemangioma; Hypermethylation; Immunoprecipitation; Ions; Knockout Mice; Lead; Life; MAP Kinase Gene; MAPK3 gene; MCAM gene; Malignant Neoplasms; Mediating; Mediator of activation protein; Mesenchymal Stem Cells; Methylation; Mus; Myocardial Infarction; Neonatal; Nuclear Translocation; Osteocytes; Pathogenesis; Pathway interactions; Phosphotransferases; Play; Population; Pregnancy; Process; Protein Binding; Protein Isoforms; Protein Kinase C; Proteins; Proto-Oncogene Proteins B-raf; Regenerative Medicine; Regulation; Reporter Genes; Role; Signal Transduction; Smooth Muscle Myocytes; Stat3 protein; Stem cells; Stroke; Structure; TP53 gene; Testing; Transcript; Transferase; Tube; Vascular Diseases; Vascular Proliferation; Vascular System; Vascular blood supply; Woman; Work; Wound Healing; angiogenesis; base; blood vessel development; crosslink; enzyme activity; epigenetic regulation; fetal; fetal blood; gain of function; genome wide methylation; innovation; loss of function; member; new therapeutic target; novel; organ growth; pluripotency; promoter; raf Kinases; reproductive; research study; response; therapeutic target; vasculogenesis

 DESCRIPTION (provided by applicant): Raf Kinase and Epigenetic Regulation of Fetal Vascular Development. Project Summary Development of the vasculature is a dynamic process that relies on the coordinated expression of numerous genes. The factors that regulate gene expression during blood vessel development are not well defined, however. Our preliminary data and reports from others of impaired vascular development in B-Raf knockout mice have elucidated the significance of this pathway in the developing vasculature. Importantly, downstream mechanism(s) through which B-Raf regulates vascular development are not known. Based on our preliminary data, we propose that B-Raf kinases via Protein Kinase C (PKC), Signal Transducer and Activator of Transcription 3 (STAT3) and p53 regulate DNA methyl Transferase (DNMT) and Ten-Eleven Translocation Methylcytosine Dioxygenase (TET) gene expression and activities. The altered activities of DNMT and TET lead to changes in promoter DNA methylation resulting in altered expression of genes regulating apoptosis such as Bcl2. These pathways play a crucial role in fetal vascular development. In this project, we will test the hypothesis that B-Raf hypomethylates the Bcl2 promoter to regulate fetal vascular stem cell survival. In the three Specific Aims, we will examine the (1) downstream mediators of B-Raf such as PKC, STAT3, and p53 (2) B-Raf-mediated regulation of DNMT and TET expression and activity, and (3) B-Raf-mediated regulation of promoter DNA methylation of the antiapoptotic gene Bcl2 via DNMT and TET. These studies are highly innovative and significant. Kinase-mediated regulation of epigenetic phenomenon such as DNA methylation is not well studied and is of fundamental importance from a basic science perspective. Moreover, pathways regulating fetal VSC apoptosis are involved in the pathogenesis of several fetal and neonatal disorders such as growth restriction, cerebral palsy, hemangioma, etc. From a clinical perspective, these studies will provide several therapeutic targets to regulate angiogenesis and vascular proliferation in many conditions from organ growth and wound healing to disorders such as myocardial infarction, cerebrovascular accident (stroke), and fetal development in general.

 描述（由申请人提供）：Raf 激酶和胎儿血管发育的表观遗传调节。项目摘要 脉管系统的发育是一个动态过程，依赖于众多基因的协调表达。然而，在血管发育过程中调节基因表达的因素尚未明确定义。我们的初步数据和其他人关于 B-Raf 敲除小鼠血管发育受损的报告阐明了该途径在发育中的脉管系统中的重要性。重要的是，B-Raf 调节血管发育的下游机制尚不清楚。根据我们的初步数据，我们提出 B-Raf 激酶通过蛋白激酶 C (PKC)、信号转导器和转录激活剂 3 (STAT3) 和 p53 调节 DNA 甲基转移酶 (DNMT) 和 10-11 易位甲基胞嘧啶双加氧酶 (TET) 基因表达和活性。 DNMT 和 TET 活性的改变导致启动子 DNA 甲基化的变化，从而导致调节细胞凋亡的基因（例如 Bcl2）的表达发生改变。这些途径在胎儿血管发育中起着至关重要的作用。在这个项目中，我们将测试 B-Raf 使 Bcl2 启动子低甲基化以调节胎儿血管干细胞存活的假设。在三个具体目标中，我们将研究 (1) B-Raf 的下游介质，如 PKC、STAT3 和 p53 (2) B-Raf 介导的 DNMT 和 TET 表达和活性调节，以及 (3) B-Raf 介导的通过 DNMT 和 TET 调节抗凋亡基因 Bcl2 的启动子 DNA 甲基化。这些研究具有很强的创新性和意义。激酶介导的表观遗传现象（例如 DNA 甲基化）的调节尚未得到充分研究，但从基础科学的角度来看具有根本重要性。此外，调节胎儿VSC凋亡的途径涉及多种胎儿和新生儿疾病的发病机制，如生长受限、脑瘫、血管瘤等。从临床角度来看，这些研究将为从器官生长和伤口愈合到心肌梗死、脑血管病等多种疾病中调节血管生成和血管增殖提供多种治疗靶点。 事故（中风）和胎儿的一般发育。