Cardiovascular Impact of Gata4 Loss in the c-Kit Lineage

c-Kit 谱系中 Gata4 缺失对心血管的影响

• 批准号: 9555818
• 负责人: Bryan D Maliken
• 金额: $ 4.7万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2021-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9555818
• 关键词: Address; Adult; Apoptosis; Appearance; Area; Autologous; Blood Vessels; Bone Marrow; Bone Marrow Transplantation; Cardiac; Cardiac Myocytes; Cardiovascular system; Cause of Death; Cell Lineage; Cell Therapy; Cell Transplantation; Cells; Cicatrix; Clinical; Clinical Trials; Coronary; Coronary artery; Development; Echocardiography; Endothelial Cells; Endothelium; Enterobacteria phage P1 Cre recombinase; Epidemic; Evaluation; Event; Flow Cytometry; GATA4 gene; Gene Expression; Generations; Genetic; Goals; Heart; Heart Injuries; Homeostasis; Human; Immunofluorescence Immunologic; In Vitro; Incentives; Individual; Infarction; Infiltration; Infusion procedures; Injury; Investigation; Ischemia; Knock-in Mouse; Knock-out; Link; Mediating; Modeling; Monitor; Mus; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardium; Natural regeneration; Physiological; Population; Prevention; Proto-Oncogene Protein c-kit; Public Health; Publishing; Receptor Protein-Tyrosine Kinases; Recovery; Reperfusion Injury; Reperfusion Therapy; Reporting; Research Personnel; Role; Site; Source; Stem cells; Supporting Cell; Therapeutic; Time; Transcriptional Regulation; Tube; Vascular Endothelium; Vascular Permeabilities; Vascularization; Vision; Work; angiogenesis; artery infusion; bone; cardiac regeneration; cardiac repair; experimental study; functional improvement; global health; heart dimension/size; heart function; improved; in vivo; insight; migration; neovascularization; new therapeutic target; novel; prophylactic; regenerative; repaired; stem; transcription factor

Project Summary Ischemic heart disease is the leading cause of death worldwide. To approach this major public health epidemic, two prevailing areas of study have emerged: 1) Cell therapy aimed at delivering culture-expanded pluripotent cells by coronary artery infusion after an ischemic event, and 2) Endogenous cardiac progenitor cells (CPCs) and their potential role for in vivo renewal and regeneration of the myocardium. Initially these two topics seemed inextricably linked, but it is now becoming clear that while delivery of CPCs for therapy may provide some mechanistically-unexplained benefit, the retention of these cells into the myocardium is marginal and de novo cardiomyocyte differentiation is even less common. A population of putative CPCs, marked by the tyrosine kinase receptor, c-Kit, have been a common focus in both areas of study. Our lab recently published a genetic lineage tracing model demonstrating the cardiac contribution of the Kit lineage in the heart which showed conclusively that Kit-derived cells rarely become cardiomyocytes, but have a high propensity for the formation of vascular endothelium. Furthermore, this study demonstrated that approximately 80% of Kit lineage-traced cardiomyocytes were actually derived from fusion events and the sought-after de novo cardiomyocyte was even more rare than originally believed. To validate this study, we aimed to eliminate the ability of Kit-derived cells to make any de novo cardiomyocytes by knocking out essential cardiomyogenic transcription factor, Gata4, theoretically allowing only for lineage-traced cardiomyocytes derived from fusion. Interestingly, the appearance of Kit lineage-traced cardiomyocytes due to fusion increased dramatically with the loss of Gata4 in the Kit lineage, and, furthermore, we observed a dramatic increase of both Kit-lineage cells in the heart and total cardiac vascular endothelium. We now hypothesize that what is uniting these findings is a novel role for Gata4 in the adult vascular endothelium, and what initially appeared to be an interesting driver of innate fusigenic character with cardiomyocytes is most likely an incidental readout due to increased vascular permeability and neoangiogenesis. While many GATA transcription factors have had previously reported roles in vascular development, including Gata4, few studies have focused on adult vascular homeostasis. Better understanding of this Gata4-mediated transcriptional control of vascular expansion could provide meaningful insight into mechanisms of cardiac renewal and repair. To address this novel role of Gata4 in the cardiac vascular endothelium, we will pursue the following aims: (1) Determine whether endothelial-specific loss of Gata4 leads to increased vascular permeability and enhanced angiogenesis; (2) Assess changes in cardiac recovery after deletion of Gata4 in the adult cardiac endothelium.

项目摘要 缺血性心脏病是全球死亡的主要原因。为了接近这种主要的公共卫生流行病， 已经出现了两个主要的研究领域：1）旨在提供培养膨胀多能的细胞疗法 缺血性事件后通过冠状动脉输注的细胞，2）内源性心脏祖细胞（CPC） 以及它们在体内更新和心肌再生中的潜在作用。最初，这两个主题似乎是 密不可分的链接，但现在越来越清楚，尽管CPC进行治疗可能会提供一些 机械上没有解释的益处，这些细胞保留到心肌是边缘和从头开始的 心肌细胞分化甚至不那么普遍。以酪氨酸激酶为特征的推定CPC人群 受体，C-KIT在两个研究领域都是普遍的重点。我们的实验室最近发表了遗传谱系 追踪模型，证明了心脏中套件谱系的心脏贡献，该模型最终显示 该基特衍生的细胞很少成为心肌细胞，但具有很高的血管形成倾向 内皮。此外，这项研究表明，大约80％的KIT谱系跟踪的心肌细胞 实际上是源自融合事件，而抢手的从头心肌细胞比 最初相信。为了验证这项研究，我们旨在消除盒子衍生细胞使任何DE的能力 Novo心肌细胞通过淘汰必需的心肌生成转录因子GATA4，理论上允许 仅适用于从融合中得出的谱系追踪的心肌细胞。有趣的是，套件谱系跟踪的外观 由于融合而导致的心肌细胞随着套件谱系中的GATA4损失而急剧增加，此外， 我们观察到了心脏中两个试剂盒细胞和总心脏血管内皮的急剧增加。 现在，我们假设将这些发现团结起来是GATA4在成年血管内皮中的新作用， 最初看来是具有心肌细胞先天熔融特征的有趣驱动力 由于血管通透性增加和新血管生成的增加，可能是偶然的读数。而许多gata 转录因子以前曾报道过血管发育中的作用，包括GATA4，很少的研究 专注于成人血管稳态。更好地了解该GATA4介导的转录控制 血管膨胀可以为心脏更新和修复机制提供有意义的见解。到 解决GATA4在心脏血管内皮中的新作用，我们将追求以下目的：（1） 确定GATA4的内皮特异性损失是否导致血管通透性增加并增强 血管生成； （2）评估成人心脏内皮中GATA4缺失后心脏恢复的变化。