Investigating the contributions of neural crest to adult regeneration

项目总结/摘要： 心脏神经嵴细胞对心血管系统的发育至关重要， 成年心脏的一小部分心肌细胞。这种独特的神经嵴衍生的 心肌细胞代表了能够控制心肌再生的心肌样细胞的吸引人的来源。 我发表的和初步的数据表明，在损伤后，sox 10+细胞在损伤诱导的增殖中扩增， 的方式，并通过启动发育中的神经嵴基因调控成功再生所需的 网络虽然初步结果表明，这些细胞很可能来自神经嵴起源，但这 还有待于更深入的探索以前对心脏再生的研究主要集中在中胚层- 现在，关于外胚层衍生的神经嵴的作用还有很多有待发现的地方。 修复过程中的细胞有了这个独立之路奖，我将寻求了解潜在的 用细胞生物学方法研究神经嵴对成人心脏修复的作用机制 技术，以及再生心脏的高分辨率成像。本提案的总体目标是 测试神经嵴来源的细胞是斑马鱼心脏再生所必需的假设（目的1），使用 单细胞RNA-seq和ChIP-seq来寻找表达不同的神经嵴来源的细胞亚群， 基因签名响应损伤（目标2），并调查，在高分辨率，形态发生事件， 允许成体sox 10+细胞参与损伤反应（Aim 3）。这一提议的核心假设是 神经嵴来源的细胞通过重新部署发育中的神经细胞， 波峰程序。这个项目的长期目标是了解开发程序是如何被重用的， 以及我们如何劫持这些基因调控回路来重新编程细胞类型 不能再生的细胞才能有更多的再生潜能。本途径建议开展的工作， 我的多学科咨询委员会将大大促进独立奖的提案， 组织再生、单细胞RNA-seq方法和分析以及基因组生物学方面的专业知识。后 发展一个强大的技能和研究基础所提供的两年的辅导阶段，这 我的目标是建立一个高影响力的独立研究小组，将联合收割机系统级 用最先进的细胞和发育生物学技术来回答再生问题 通过发展的透镜。长期项目假设的重点是我们如何能够再生人类 心我的初步数据显示，心脏神经嵴细胞不仅对鱼类的心肌细胞有贡献， 因此，刺激这些细胞促进斑马鱼再生的机制可能 提供治疗方法来修复包括人类在内的哺乳动物的心脏损伤。

PROJECT SUMMARY/ ABSTRACT: Cardiac neural crest cells are crucial for the development of the cardiovascular system by contributing to a small proportion of cardiomyocytes of the adult heart. This unique population of neural crest-derived cardiomyocytes represents an appealing source of precursor-like cells able to control myocardial regeneration. My published and preliminary data show that upon injury sox10+ cells expand in an injury-induced proliferative manner and are required for successful regeneration by initiating a developing neural crest gene regulatory network. While preliminary results suggest it is likely these cells are derived from a neural crest origin, this has yet to be explored at great depth. Previous studies of cardiac regeneration have focused primarily on mesoderm- derived populations, and, now, much remains to be uncovered about the role of ectoderm-derived neural crest cells in the repair process. With this Pathway to Independence Award, I will seek to understand the underlying mechanisms of neural crest contributions to adult heart repair using cell biological approaches, systems-level techniques, and high-resolution imaging of regenerating hearts. The overall objectives of this proposal are to test the hypothesis that neural crest-derived cells are required for heart regeneration in zebrafish (Aim 1), use single cell RNA-seq and ChIP-seq to look for subpopulations of neural crest-derived cells that express distinct gene signatures in response to injury (Aim 2), and investigate, at high-resolution, the morphogenetic events that allow adult sox10+ cells to contribute to the injury response (Aim 3). The central hypothesis of this proposal is that neural crest-derived cells play an essential role in heart regeneration by redeploying a developmental neural crest program. The long term goal of this project is to understand how developmental programs are reused for regeneration in the adult body and how we can hijack these gene regulatory circuits for reprogramming cell types that do not regenerate in order to have a more regenerative potential. The work proposed in this Pathway to Independence Award proposal will be greatly facilitated by my multi-disciplinary advisory committee with expertise in tissue regeneration, single cell RNA-seq approaches and analyses, and genome biology. After developing a formidable skillset and research foundation afforded by the two years of the mentored phase of this award, my goal is to establish a high impact, independent research group that will combine systems-level approaches with state-of-the-art cell and developmental biology techniques to answer questions of regeneration through a developmental lens. Long term project hypotheses are focused on how we can regenerate the human heart. My preliminary data shows cardiac neural crest cells contribute to cardiomyocytes not only in fish but also in amniotes; therefore, the mechanisms that stimulate these cells to contribute to regeneration in zebrafish may offer therapeutic approaches to repair heart damage in mammals including humans.