Chemical Genetics of Vertebrate Vascular Development

描述（由申请人提供）： 了解脊椎动物的循环系统是如何发展的是一个基本的生物学挑战与巨大的临床意义。 斑马鱼的分子和遗传学研究突出了血管内皮生长因子（VEGF），Notch和gridlock（grl）在胚胎血管发育信号通路中的关键作用。斑马鱼已被证明非常适合化学遗传学分析，其中通过高通量，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。 种子工作最近发现了一种新的化合物，GS 4012，凭借其抑制grl基因突变引起的血管缺陷的能力。 grl抑制剂筛选还鉴定了另一种抑制剂，GS 4898，其具有与GS 4012完全不同的结构，并且可能具有不同的作用机制。 本研究采用分子和化学遗传学方法研究脊椎动物血管发育，具体目标有三：1）确定GS 4012如何挽救突变胚胎中的血管堵塞缺陷。 2)鉴定培养的内皮细胞中GS4012靶向的信号通路。 3)确定GS 4898在体内和体外的作用机制。 该项目将有助于回答有关血管发生的一些悬而未决的问题，并为进一步研究提供一套有价值的化学工具。 像GS 4012和GS 4898这样调节血管发育的电网抑制剂，可以作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MGH心血管研究中心的独特资源来扩展他的科学技能。 他的两个赞助商将指导他在化学遗传学，斑马鱼生物学，心血管细胞生理学，以及在专业发展。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成血管细胞发育领域的领导者。 GS 4012、GS 4898和其他gridlock抑制剂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独立研究者。

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.