Elucidating the mechanisms of venous reprogramming during coronary development

描述（由申请人提供）：冠心病是全世界死亡的主要原因，每年造成数百万人死亡，而数千万人通过药物或手术干预来控制疾病。这种疾病是由冠状动脉的病理学引起的，但尽管它对全球医疗保健产生了影响，但令人惊讶的是，人们对冠状动脉的发展知之甚少，这可能会阻碍改善现有临床治疗的努力。我研究的长期目标是以高分辨率描述胚胎发生期间从祖细胞产生冠状动脉的所有细胞分化和形态发生步骤。我最近确定了这些祖细胞在小鼠中的位置，并描述了导致成熟冠状动脉的发育途径。该途径中的每一步都发生在固定的位置，这表明位置特异性线索触发了特定的细胞分化事件。冠状血管开始为静脉窦的静脉芽，当它们迁移到心脏上并侵入心脏时去分化。随后，血管根据其在心脏内的位置分化成动脉、毛细血管或静脉。在这个提议中，我们将调查的初始步骤，发芽和静脉重编程，通过执行以下目标：（1）通过使用微阵列来表征位于这些事件发生的细胞的转录谱，以确定候选发芽和去分化诱导剂。(2)通过在体外功能测定中测试从这些细胞中分离的蛋白质来鉴定诱导内皮去分化的因素。(3)通过评估基因敲除动物或用功能阻断抗体或药物治疗的动物的冠状血管发育，测试目标1和2的候选物在体内的作用。该提案的最初部分将在斯坦福大学的Mark Krasnow博士的实验室进行，该实验室的环境支持了许多创新研究项目，这些项目带来了富有成效的独立职业生涯。在这里，我将实现我的直接目标，即找到一个终身职位，以完成该提案的目标。在结论中，我们将确定冠状动脉发育的第一步的机制，提供对这些新过程的洞察，以及提供一个实验模型来描述路径的其余部分。我们也将更进一步了解如何异位诱导冠状动脉血管，这应该是有价值的信息，在进一步的临床发展。 公共卫生相关性：冠状动脉疾病是世界范围内死亡的主要原因，对于数千万通过药物或外科手术治疗其症状的人来说，这是一个重大的医疗负担。该提案概述了旨在寻找诱导胚胎冠状动脉形成的蛋白质的实验，以便它们可用于指导治疗方法的开发，例如用于搭桥手术或血管重建药物的工程血管。

DESCRIPTION (provided by applicant): Coronary heart disease is the leading cause of death worldwide killing millions per year while tens of millions manage the disease medically or by undergoing surgical interventions. The disease results from pathology of the coronary arteries, but despite its impact on worldwide healthcare, surprisingly little is known about coronary artery development, which likely hampers efforts to improve available clinical treatments. The long-term goal of my research is to describe, at high resolution, all the cell differentiation and morphogenesis steps that produce coronary arteries from their progenitor cells during embryogenesis. I have recently identified the location of these progenitors in mice and described the developmental pathway leading to mature coronary arteries. Each step in the pathway occurred at stereotyped locations suggesting that position- specific cues trigger specific cell differentiation events. Coronary vessels begin as venous sprouts of the sinus venosus that de-differentiate as they migrate onto and invade the heart. Subsequently, the vessels differentiate into arteries, capillaries, or veins depending on their location within the heart. In this proposal, we will investigate the initial steps, sprouting and venous reprogramming, by carrying out the following aims: (1) Identify candidate sprouting and de-differentiation inducers by using microarrays to characterize the transcriptional profiles of cells located where these events occur. (2) Identify factors that induce endothelial de-differentiation by testing proteins isolated from these cells in an in vitro functional assay. (3) Test candidates from aims 1 and 2 for their role in vivo by assessing coronary vessel development in knockout animals or those treated with function-blocking antibodies or drugs. The initial part of the proposal will be carried out at Stanford University in Dr. Mark Krasnow's lab, an environment that has supported many innovative research projects that have led to fruitful independent careers. Here, I will carry out my immediate goal of finding a tenure- track position in which to complete the proposal's aims. At the conclusion, we will have identified the mechanisms underlying the first steps of coronary artery development providing insight into these novel processes as well as providing an experimental model for describing the remaining portions of the pathway. We will also be a step closer in understanding how to ectopically induce coronary vessels, which should be valuable information in furthering clinical developments. PUBLIC HEALTH RELEVANCE: Coronary artery disease is the leading cause of death worldwide and a significant healthcare burden for tens of millions who manage its symptoms medically or with surgical interventions. This proposal outlines experiments aimed at finding the proteins that induce coronary artery formation in the embryo so that they can be used to guide the development of therapeutics such as engineered vessels for bypass surgery or revascularization drugs.