Investigating the contributions of neural crest to adult regeneration

项目摘要/摘要： 心脏神经脊细胞对心血管系统的发育至关重要，因为它有助于 成人心脏的心肌细胞比例较小。这一独特的神经脊群起源于 心肌细胞是控制心肌再生的前体细胞的诱人来源。 我已发表的初步数据显示，在损伤后，Sox10+细胞以损伤诱导的增殖方式扩张 启动发育中的神经脊基因调控，是成功再生所必需的 网络。虽然初步结果表明这些细胞很可能来自神经脊来源，但这已经 还有待深入探索。以前对心脏再生的研究主要集中在中胚层- 外胚层衍生的神经脊的作用目前仍有许多未被发现的地方。 处于修复过程中的细胞。有了这个独立之路奖，我将试图理解潜在的 用细胞生物学方法在系统水平上研究神经脊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。这项建议的总体目标是 测试关于斑马鱼心脏再生需要神经脊源性细胞的假设(目标1)，使用 单细胞RNA-SEQ和ChIP-SEQ寻找表达不同基因的神经脊来源细胞亚群 对伤害作出反应的基因签名(目标2)，并在高分辨率下调查 允许成年SOX10+细胞参与损伤反应(目标3)。这项提议的中心假设是 神经沟来源的细胞通过重新部署发育中的神经在心脏再生中发挥重要作用 CREST程序。这个项目的长期目标是了解发展项目是如何被重复使用的 成人身体的再生，以及我们如何劫持这些基因调控电路来重新编程细胞类型 它们不会再生，以获得更大的再生潜力。在这条道路上提出的工作是 我的多学科咨询委员会将极大地促进独立奖的提案 在组织再生、单细胞RNA序列方法和分析以及基因组生物学方面的专业知识。之后 发展强大的技能和研究基础，这是本课程指导阶段的两年 获奖，我的目标是建立一个高影响力的、独立的研究小组，将系统级的 用最先进的细胞和发育生物学技术回答再生问题的方法 通过发育透镜。长期项目假设的重点是我们如何重生人类 心。我的初步数据显示，心脏神经脊细胞不仅对鱼类的心肌细胞有贡献，而且对心肌细胞也有贡献 因此，刺激这些细胞促进斑马鱼再生的机制可能 提供修复包括人类在内的哺乳动物心脏损伤的治疗方法。

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.