iPS cells-derived progenitor cells for angiomyogenesis

描述（由申请人提供）：已采用各种组织工程策略来治疗心肌梗死导致的受损心肌。然而，重建的心肌必须包括能够在不同代谢需求下滋养它的血管网络。最近，我们证明了这种方法的有效性，使用从遗传修饰的间充质干细胞开发的心脏组织移植物。虽然证明了这种方法的价值，但用于组织工程的更好的细胞来源是来自患者自身组织的诱导多能干细胞（iPSC）。我们的目标是开发一种新的方法来治疗心肌梗死（MI），使用啮齿动物模型，涉及使用iPSCs的组织工程。这种方法最终可以用于人类，并允许自体移植，从而消除宿主排斥反应的问题。iPSC的使用将允许有效分化为内皮细胞和其他心脏谱系细胞，供体和受体组织之间的免疫相容性，以及营养物质和废物快速运输到新的和发育中的组织（通过血管形成）。我们假设，由iPSC衍生的内皮细胞和其他心脏谱系细胞的网络组成的三细胞补片将有效地用于新血管和心肌组织的再生长，这反过来可能导致心脏功能的改善。在目标1中，我们将进行体外研究，以表征iPSC分化，并确定其定向分化为内皮细胞和心肌细胞谱系的最佳条件，这些细胞谱系将适用于开发用于心脏修复的三细胞贴片。我们将进一步增强心脏前体细胞的血管生成或肌生成潜力，并确定预处理是否促进iPSC分化为心脏谱系细胞。在目标2中，我们将确定预血管化的细胞贴片是否会增加植入后iPSC衍生的心脏表型的保留和存活，从而导致血管分布、灌注和心脏功能的显著改善。目的3中的研究将确定通过腺苷酸环化酶或特异性纤维化抑制性microRNA的过度表达操纵亚细胞途径来下调纤维化是否会影响心肌梗死后心脏祖细胞贴片的植入。这些研究将为开发工程iPSC衍生的心脏组织补片作为心肌再生的可行疗法提供新的见解。 公共卫生相关性：本项目将研究由内皮细胞、肌细胞和其他来源于诱导多能干细胞（iPSC）的心脏谱系细胞组成的预血管化三细胞补片在体内应用于梗死心脏后的存活和植入的有效性。

DESCRIPTION (provided by applicant): Various tissue engineering strategies have been adopted to treat damaged heart muscle as a result of myocardial infarction. However, the rebuilt myocardium must include a vascular network able to nourish it under diverse metabolic demands. Recently we demonstrated the efficacy of such an approach using cardiac tissue grafts developed from genetically modified mesenchymal stem cells. While demonstrating the value of such an approach, a better source of cells for tissue engineering would be induced pluripotent stem cells (iPSCs) derived from the patient's own tissues. Our objective is to develop a novel approach to treatment of myocardial infarction (MI), using a rodent model, that involves tissue engineering using iPSCs. Such an approach could eventually be used in humans and would allow autologous transplantation, thereby eliminating the problem of host rejection. The use of iPSCs will allow efficient differentiation into endothelial cells and other cardiac lineage cells, immune compatibility between donor and recipient tissues, and rapid transport of nutrients and waste products to new and developing tissue (via blood vessel formation). We hypothesize that a tri-cell patch composed of a network of iPSC-derived endothelial cells and other cardiac lineage cells will be effective for regrowth of neovasculature and myocardial tissue, which in turn could lead to improved cardiac function. In Aim 1, we will perform in vitro studies to characterize iPSC differentiation and define the optimal conditions for their directed differentiation into endothelial and cardiomyocyte cell lineages that will be suitable for development of a tri-cell patch to be used in cardiac repair. We will further enhance the angiogenic or myogenic potential of cardiac precursor cells, and determine whether preconditioning promotes differentiation of iPSCs into cardiac lineage cells. In Aim 2 we will determine whether a prevascularized cell patch will increase the retention and survival of iPSC-derived cardiac phenotypes after implantation leading to significant improvements in vascularity, perfusion, and cardiac function. Studies in Aim 3 will determine whether downregulation of fibrosis by manipulating subcellular pathways by over expression of adenylyl cyclases or specific fibrosis repressive microRNAs will influence the engraftment of a cardiac progenitor cell patch after myocardial infarction. These studies will provide new insights into the development of engineered iPSC-derived cardiac tissue patches as a viable therapy for cardiac muscle regeneration. PUBLIC HEALTH RELEVANCE: This project will investigate the effectiveness of prevascularized tri-cell patch composed of endothelial cells, myocytes, and other cardiac lineage cells derived from induced pluripotent stem cells (iPSCs) for their survival and engraftment after in vivo application on infarcted hearts.