Therapeutic Mechanisms of Cardiac Progenitors in Ischemic Cardiomyopathy

心脏祖细胞在缺血性心肌病中的治疗机制

基本信息

批准号：
9220049
负责人：
Kenneth Michael Fish
金额：
$ 84.07万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-01-20 至 2020-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9220049
关键词：
Address Animal Model Animals Biological Response Modifier Therapy Biology Cardiac Cardiac Myocytes Cardiovascular system Cause of Death Cell Count Cell Therapy Cell Transplantation Cell Transplants Cell physiology Cells Clinical Data Discipline Endothelial Cells Family suidae Foundations Gene Transfer Genes Genomics Heart Heart failure Human Imaging Techniques In Vitro Injury Lead Ligands Literature Mediating Membrane Messenger RNA MicroRNAs Modeling Molecular Myocardial Myocardial Ischemia Natural regeneration PDGFRB gene Platelet-Derived Growth Factor alpha Receptor Play Positioning Attribute Pre-Clinical Model Publishing Rattus Recovery Recruitment Activity Regenerative Medicine Research Research Personnel Role Route Source Stem Cell Factor Stem cells Technology Testing Therapeutic Treatment Efficacy Tyrosine angiogenesis cardiac repair cell type clinically relevant design exosome experimental study gain of function improved innovation insight ischemic cardiomyopathy knock-down loss of function nanoparticle next generation novel novel therapeutic intervention paracrine pre-clinical progenitor programs receptor regenerative repaired response restoration small hairpin RNA success tissue repair vector

项目摘要

Ischemic heart disease is a leading cause of death worldwide. Given the limited regenerative capacity of the human heart following myocardial injury, stem cell-based therapies, including the recent cKit+ cell therapy have emerged as a promising approach for improving cardiac repair and function. Clinical and preclinical data suggests that benefits of cKit+ cell transplantation include improved cardiac function and myocardial mass. However, the ability of cKit+ CPCs to differentiate into cardiomyocytes as a significant mode of cardiac repair is controversial. Published data from our group has established the beneficial effects of stem cell factor (SCF), the ligand for cKit, in improving the abundance of cKit+ cells, angiogenesis and cardiac function in both rat and pig models of ischemic cardiomyopathy. Further, our preliminary data suggest that secreted paracrine factors, particularly the membrane-bound exosomes, may mediate most of the pro-angiogenic paracrine activity of the SCF-treated cKit+ cell. Our central hypothesis is that mobilization of a heterogeneous group of endogenous CPCs using gene transfer for the receptor tyrosine cKit ligand stem cell factor (SCF) can enhance cardiac tissue repair following myocardial injury via their paracrine secretion. We will test the necessary and sufficient conditions of SCF-induced CPC recruitment using the MI model of heart failure in swine to evaluate the roles of SCF-induced cKit, PW1 and PDGFR expressing CPCs in myocardial repair, regeneration and restoration of function. We will test the hypothesis that cKit, PW1 and PDGFR progenitor cells are sufficient for SCF induced cardiac repair by gain of function experiments using SCF modRNA. We will test the hypothesis that cKit, PW1 and PDGFR progenitor cells are necessary for cardiac repair by loss of function experiments using shRNA-mediated knockdown of cKit, PDGFR and PW1 receptors. RNAs will be delivered one-week post-MI by the clinically relevant intracoronary route and animals will be evaluated for survival, cardiac remodeling, cardiac function, cell-specific proliferation and differentiation and the molecular mechanisms involved. We will determine the role of paracrine secretion from cKit+ CPCs in SCF-induced cardiac repair. Our preliminary data suggest that exosomes, and not the exosomes-depleted fraction from swine post-MI cKit- secretome have pro-angiogenic activity. Using specific expertise for exosomes research, we will identify the miRNA expression of cKit+ exosomes in response to SCF. In addition, we will investigate the functional mechanisms of cKit+ exosomes-induced proliferation and contractile function of cardiomyocytes and angiogenesis of endothelial cells in vitro. Finally, we will determine the therapeutic benefits of cKit exosomes in a swine model of heart failure.

鉴于再生能力有限，缺血性心脏病是全世界死亡的主要原因。心肌损伤后的人类心脏，基于干细胞的疗法，包括最近的 cKit+ 细胞疗法已经正在成为改善心脏修复和功能的一种有前景的方法。表明 cKit+ 细胞移植的好处包括改善心脏功能和心肌质量。然而，cKit+ CPC 分化为心肌细胞作为心脏修复的重要模式的能力是我们小组发表的数据已经证实了干细胞因子（SCF）的有益作用，即干细胞因子（SCF）。 cKit 配体，可改善大鼠和猪的 cKit+ 细胞丰度、血管生成和心脏功能此外，我们的初步数据表明，分泌的旁分泌因子，特别是膜结合的外泌体，可能介导大部分促血管生成旁分泌活性 SCF 处理的 cKit+ 细胞的中心假设是异质组的动员。使用受体酪氨酸 cKit 配体干细胞因子 (SCF) 基因转移的内源性 CPC 可以通过旁分泌分泌增强心肌损伤后心脏组织的修复。我们将使用 MI 模型来测试 SCF 诱导的 CPC 招募的充分必要条件猪心力衰竭的研究，以评估 SCF 诱导的 cKit、PW1 和 PDGFRα 表达 CPC 在猪心力衰竭中的作用心肌修复、再生和功能恢复我们将检验 cKit、PW1 和功能的假设。通过功能获得实验，PDGFR祖细胞足以进行 SCF 诱导的心脏修复我们将检验 cKit、PW1 和 PDGFR 祖细胞对于 SCF modRNA 所必需的假设。使用 shRNA 介导的 cKit、PDGFRα 和 PW1 敲低进行功能丧失实验的心脏修复 RNA 将在 MI 后一周通过临床相关的冠状动脉内途径和动物递送。将评估存活率、心脏重塑、心脏功能、细胞特异性增殖和分化以及所涉及的分子机制。我们将确定 cKit+ CPC 旁分泌在 SCF 诱导的心脏修复中的作用。初步数据表明，外泌体，而不是 MI 后的猪的外泌体耗尽部分 cKit- 分泌组具有促血管生成活性。利用外泌体研究的特定专业知识，我们将确定 cKit+ 外泌体响应 SCF 的 miRNA 表达此外，我们将研究其功能。 cKit+外泌体诱导心肌细胞增殖和收缩功能的机制最后，我们将确定 cKit 外泌体在体外的治疗效果。猪心力衰竭模型。