Chemical Genetics of Vertebrate Vascular Development

描述(由申请人提供)： 了解脊椎动物循环系统是如何发展的是一个具有巨大临床意义的基本生物学挑战。斑马鱼的分子和遗传学研究强调了血管内皮生长因子(VEGF)、Notch和GRL在胚胎血管发育信号通路中的关键作用。斑马鱼已经被证明非常容易接受化学遗传学分析，在化学遗传学分析中，通过高通量、基于表型的筛选发现的小分子被用作解剖细胞和发育过程的工具。开创性的工作最近发现了一种新的化合物，GS4012，凭借其抑制gr1基因突变引起的血管缺陷的能力。GRL抑制子筛选还发现了另一个抑制子GS4898，它具有与GS4012完全不同的结构，并可能具有不同的作用机制。这项建议采用分子和化学遗传学的方法来研究脊椎动物的血管发育，有三个具体的目的：1)确定GS4012如何挽救突变胚胎中的僵局血管缺陷。2)确定GS4012在培养内皮细胞中的靶向信号通路。3)明确GS4898的体内外作用机制。这个项目将有助于回答一些关于血管生成的悬而未决的问题，并为进一步研究提供一套有价值的化学工具。GS4012和GS4898这样的堵塞抑制因子可以调节血管发育，可能成为治疗缺血性血管疾病等重要疾病的先导化合物。 这位候选人拥有分子生物学和发育遗传学的背景，最近完成了心脏病学奖学金培训。洪博士现在寻求通过麻省理工学院心血管研究中心提供的独特资源来扩展他的科学技能。他的两个赞助商将在化学遗传学、斑马鱼生物学和心血管细胞生理学以及职业发展方面指导他。洪博士的顾问小组包括血管生成、斑马鱼器官发生和斑马鱼血管母细胞发育领域的领军人物。对GS4012、GS4898和其他堵塞抑制因子的作用机制研究将成为进一步科学发展的跳板，最终使洪博士成为基础血管生物学领域的一名独立研究员。

DESCRIPTION (provided by applicant): Understanding how the vertebrate circulatory system develops is a fundamental biological challenge with tremendous clinical implications. Molecular and genetic studies in zebrafish have highlighted the key roles of vascular endothelial growth factor (VEGF), Notch, and gridlock (grl) in the signaling pathway for embryonic vascular development. The zebrafish has proven to be remarkably amenable to chemical genetics analysis, in which small molecules discovered by high-throughput, phenotype-based screens are used as tools for dissecting cellular and developmental processes. Seminal work has recently identified a novel compound, GS4012, by virtue of its ability to suppress the vascular defect caused by a mutation in the grl gene. The grl suppressor screen has also identified another suppressor, GS4898, which has a completely different structure, and presumably distinct mechanism of action, from GS4012. This proposal takes molecular and chemical genetic approaches to study vertebrate vascular development, with three specific aims: 1) Determine how GS4012 rescues the gridlock vascular defect in mutant embryos. 2) Identify signaling pathways targeted by GS4012 in cultured endothelial cells. 3) Identify the mechanism of action of GS4898 in vivo and in vitro. This project will help to answer some of the outstanding questions regarding vasculogenesis and provide a valuable set of chemical tools for further study. Gridlock suppressors like GS4012 and GS4898, which modulate vascular development, may serve as lead compounds for treatments of important illnesses such ischemic vascular diseases. The candidate, who has a background in molecular biology and developmental genetics, has recently completed cardiology fellowship training. Dr. Hong now seeks to expand upon his scientific skills through unique resources available at the MGH's Cardiovascular Research Center. His two sponsors will guide him in chemical genetics, zebrafish biology, and cardiovascular cellular physiology, as well as in professional development. Dr. Hong's panel of advisors includes leaders in the fields of angiogenesis, zebrafish organogenesis, and zebrafish hemangioblast development. Mechanism of action studies of GS4012, GS4898, and additional gridlock suppressors will serve as a springboard for further scientific development, culminating in Dr. Hong's emergence as an independent investigator in the field of basic vascular biology.