Elucidating the mechanisms of venous reprogramming during coronary development

项目摘要/摘要 冠心病是全球范围内每年导致数百万人死亡的主要原因 虽然数以千万计的人通过医学或外科手术控制了这种疾病 干预措施。这种疾病是由冠状动脉的病理引起的，但尽管 它对全球医疗保健的影响，令人惊讶的是，人们对冠状动脉知之甚少 发展，这可能阻碍改进现有临床治疗的努力。这个 我研究的长期目标是以高分辨率描述所有细胞分化 以及从其祖细胞产生冠状动脉的形态发生步骤 在胚胎发育过程中。我最近确定了这些祖先的位置 并描述了导致冠状动脉成熟的发育途径。 通路中的每一步都发生在刻板的位置上，这表明- 特定的线索会触发特定的细胞分化事件。冠状动脉始于 静脉窦的静脉芽，当它们迁移到和 入侵心脏。随后，血管分化为动脉、毛细血管或 静脉取决于它们在心脏内的位置。在这份提案中，我们将调查 发芽和静脉重新编程的初始步骤，通过执行以下步骤 目的：(1)筛选出候选的萌发和去分化诱导剂 微阵列用于表征位于这些位置的细胞的转录图谱 事件就会发生。(2)通过检测确定诱导内皮细胞去分化的因素 在体外功能分析中从这些细胞中分离出的蛋白质。(3)考生来自 通过评估基因敲除中冠状动脉的发育来研究它们在活体中的作用 动物或使用功能阻断抗体或药物治疗的动物。的开头部分 该提案将在斯坦福大学马克·克拉斯诺博士的实验室进行，并 支持许多创新研究项目的环境，这些项目导致了 卓有成效的独立事业。在这里，我将实现我的眼前目标，找到终身教职- 跟踪完成建议书目标的位置。在结束时，我们将拥有 确定了冠脉发育第一步的潜在机制 提供了对这些新过程的见解，并提供了一个实验模型 用于描述路径的剩余部分。我们也将更接近于 了解如何异位诱导冠状动脉血管，这应该是有价值的 进一步推动临床发展的信息。

Project Summary/Abstract 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.