Epigenomic Memory of iPSC-Derived Endothelial Cells for Cardiovascular Diseases

描述(申请人提供)：诱导多能干细胞(IPSCs)是心血管再生医学的新来源。由于IPSCs来源于受者的组织，它们避免了人类胚胎干细胞(HESCs)领域的两个主要问题，即免疫排斥和关于胚胎破坏的政治/伦理担忧。然而，目前该领域还处于初级阶段，围绕IPSC衍生品的效率、安全性和有效性的问题需要在减少到临床实践之前得到解决。这个多学科项目将研究IPSC来源的内皮细胞(IPSC-ECs)的表观基因组学，用于小鼠心肌梗死模型。首先，内皮细胞和成纤维细胞将被去分化为IPSCs。这两株细胞以及hESCs将分化为内皮细胞。干细胞、它们的起源组织(对于两个IPSC)和它们的内皮来源组织将接受第二代测序，以确定它们的表观基因组状态。最后，这三个干细胞来源的内皮细胞系将被注射到小鼠心肌梗死系统中。随着时间的推移，他们对心肌功能的影响将被观察到。这些实验有望揭示更有效的体细胞重新编程、更强大的干细胞分化和更大的心肌缺血治疗能力背后的表观基因组特征。这些研究应该为确定可以整合到IPSC-EC制剂中的分子干预铺平道路，以便最终这些干细胞衍生物可能在心血管疾病患者的临床环境中有效

DESCRIPTION (provided by applicant): Induced pluripotent stem cells (iPSCs) are a novel source for cardiovascular regenerative medicine. Because iPSCs are derived from the tissues of recipient patients, they obviate two major problems in the field of human embryonic stem cells (hESCs), namely immune rejection and the political/ethical concerns regarding the destruction of embryos. However, at present the field is in its infancy, and issues surrounding the efficiency, safety, and efficacy of iPSC-based derivatives require resolution before reduction to clinical practice occurs. This multi-disciplinary project will investigate the epigenomics of iPSC-derived endothelial cells (iPSC-ECs) for use in a murine myocardial infarction model. First, endothelial cells and fibroblasts will be de-differentiated into iPSCs. These two lines, as well as hESCs, will then be differentiating into endothelial cells. The stem cells, their tissues of origin (for the two iPSCs), and their endothelial derived tissues will be subjected to second-generation sequencing to determine their epigenomic states. Finally, the three stem cell-derived endothelial cell lines will be injected into a murine myocardial infarction system. Their effects on myocardial function will be observed over time. These experiments are anticipated to reveal the epigenomic features underlying more efficient somatic cell reprogramming, more robust stem cell differentiation, and greater therapeutic capacity for myocardial ischemia. These studies should pave the way to identifying molecular interventions which can be integrated into iPSC-EC preparation, so that eventually, these stem cell derivatives may be effective in the clinical setting for cardiovascular disease patients