The Roles of Neural Crest Derived Cardiomyocytes in Adult-Onset Heart Failure and Regeneration

项目摘要 成人发病的心肌病和心力衰竭在全世界范围内都是重大的健康和经济负担。 预计到2030年，患病率将增加46%。然而，在中国，动物模型很少 以测试成人心力衰竭的病因和可能的干预措施。这个项目创作了两部小说 斑马鱼成年性心肌病和心力衰竭模型。第一种模型利用交叉点 双转基因独家标记神经脊源性心肌细胞(NC-CMS)，一小部分 陈旧的定位于脑室的心肌细胞是正常构型所必需的 小梁，并持续整个成年生活。我们的双重转基因技术允许独家消融NC-CM 任何生命阶段的血统；胚胎发育期间的消融会导致成人心肌病和心脏 运动负荷测试失败。Notch途径配体jag2b富含NC-CMS。陷波信令是 在相邻的中胚层来源的心肌细胞中激活，而在NC-CMS中不激活。这些重要的分子 心肌细胞谱系之间的差异导致了第二种模式，即jag2b的基因突变，它具有 胚胎小梁模式缺陷和成人型心肌病。 与斑马鱼不同的是，成年哺乳动物在进化过程中失去了受伤后再生心脏的能力 明显缺乏NC-CMS，在进化上或发育上丢失。对这些问题的协同分析 斑马鱼和哺乳动物之间的进化差异将是有益的。对脑室的反应 切除，缺乏NC-CMS的成年斑马鱼无法再生正常的心脏，涉及NC-CMS和他们的心脏 成人心脏再生中的基因调控网络(GRN)一系列多方面的转基因实验 将测试胚胎来源的NC-CM谱系的再生要求和从头激活 切除心脏中的NC-cm颗粒细胞。这些结果可能导致可以挽救这种能力的途径干预 在成年哺乳动物的心脏中再生。 拟议的研究使用了两个新的斑马鱼模型和广泛的工具，包括血统和 时间调控转基因、突变体、心脏生理学、前沿成像和分子 了解成人起病心肌病、心力衰竭和心脏再生病因的方法 在斑马鱼身上。这项研究将发现使NC-CMS与众不同的发育机制和GRN 从邻近的心室肌细胞中分离出来，这是预防成人心肌病所必需的 并促进成人心脏再生。我们的目标是定位这些斑马鱼模型以测试药理作用 以及其他干预措施，以促进我们对人类心脏病的了解和治疗。

Project Summary Adult-onset cardiomyopathies and heart failure are significant health and economic burdens throughout the world, with prevalence projected to increase by 46% by 2030. However, there is a paucity of animal models in which to test the etiologies and possible interventions in adult heart failure. This project has created two novel models of adult-onset cardiomyopathy and heart failure in zebrafish. The first model utilizes intersectional double transgenics to exclusively label Neural Crest derived Cardiomyocytes (NC-Cms), a small group of cardiomyocytes that are stereotypically positioned in ventricles, are required for normal patterning of trabeculae, and persist throughout adult life. Our double transgenics allow exclusive ablation of NC-Cm lineages at any stage of life; ablation during embryogenesis results in adult-onset cardiomyopathy and heart failure in exercise stress tests. The Notch pathway ligand jag2b is enriched in NC-Cms. Notch signaling is activated in adjacent mesoderm-derived cardiomyocytes, not in NC-Cms. These important molecular distinctions between cardiomyocyte lineages led to the second model, genetic mutants of jag2b, which have both embryonic trabeculation patterning defects and adult-onset cardiomyopathy. In contrast to zebrafish, adult mammals have evolutionarily lost the ability to regenerate hearts upon injury and apparently lack NC-Cms, lost either evolutionarily or developmentally. Synergistic analysis of these evolutionary distinctions between zebrafish and mammals will be informative. In response to ventricle resection, adult zebrafish lacking NC-Cms fail to regenerate normal hearts, implicating NC-Cms and their gene regulatory networks (GRNs) in adult heart regeneration. A multifaceted series of transgenic experiments will test the regeneration requirements of embryonically-derived NC-Cm lineages and de novo activation of NC-Cm GRNs in resected hearts. These results might lead to pathway interventions that can rescue the ability to regenerate in adult mammalian hearts. The proposed research uses two novel zebrafish models and a wide variety of tools, including lineage and temporally regulated transgenics, mutants, cardiac physiological, cutting-edge imaging and molecular approaches to understand the etiologies of adult-onset cardiomyopathy, heart failure and cardiac regeneration in zebrafish. This research will discover developmental mechanisms and GRNs that make NC-Cms distinct from their neighboring ventricle cardiomyocytes, and that are required to prevent adult-onset cardiomyopathy and facilitate adult heart regeneration. Our goals are to position these zebrafish models to test pharmacological and other interventions in order to contribute to our understanding and treatments of human heart diseases.