Functional Vascular Progenitors from Naive Human iPSC

干细胞治疗儿童和成人缺血性疾病，如脑血管中风、镰状细胞病和糖尿病视网膜病变，最终不仅需要再生受损的心肌、脑、造血和视网膜组织，还需要重建引发最初疾病的有缺陷的血管生态位。人的血管系统通常起源于大量的胚胎成血管细胞或血管祖细胞（VP），在早期发育过程中分化为血管内皮细胞和周细胞。这种多产的胚胎样VP在成人中是罕见的或不存在的。此外，尽管循环成人内皮祖细胞（EPC）已被提出用于血管细胞治疗，但这种EPC不仅在多能性和扩增方面受到限制，而且在糖尿病等疾病中也存在功能缺陷。如果能够用自体或HLA匹配的胚胎血管/周细胞祖细胞有效修复缺血脱细胞毛细血管，可以逆转终末期血管疾病，如糖尿病和心脏缺血。一种解决方案是将人诱导多能干细胞（hiPSC）分化为具有高度多产的胚胎样内皮-间充质-周细胞再生潜能的VP。然而，这一目标目前受到效率低下和可变性的限制

DESCRIPTION (provided by applicant): Functional Vascular Progenitors from Na�ve Human iPSC Stem cell treatments for pediatric and adult ischemic disorders such as cerebro-vascular stroke, sickle cell disease, and diabetic retinopathy ultimately require not only the regeneration of damaged myocardium, brain, hematopoietic, and retinal tissues, but also the reconstruction of the defective vascular niche that instigated the initial disease to begin with. The human vasculature normally arises from highly prolific embryonic angioblasts or vascular progenitors (VP) that differentiate into vascular endothelial cells and pericytes during early development. Such prolific embryo-like VP are rare or non-existent in the adult. Furthermore, although circulating adult endothelial progenitor cells (EPC) have been proposed for vascular cellular therapies, such EPC are not only limited in multipotency and expansion, but also functionally defective in diseases such as diabetes. If ischemic acellular capillaries could be efficiently repaired with autologous or HLA- matched embryonic vascular/pericytic progenitors, end stage vascular diseases such as diabetes and cardiac ischemia could be reversed. One solution would be to differentiate human induced pluripotent stem cells (hiPSC) into VP that possess highly prolific embryo-like endothelial- mesenchymal-pericytic potential for regenerating diseased tissues. However, this goal is currently limited by the poor efficiency and variability of directed vascular differentiation from hiPSC. It will be necessary to first generate high-quality clinical grade hiPSC via safer non-integrating, nonviral derivation methods. In this project, we will accomplish this goal by efficiently reprogramming a patient's own blood cells to a novel high quality state of pluripotency called the "na�ve ground state" that resembles mouse embryonic stem cells (ESC). Na�ve human iPSC (N-hiPSC) possess more versatile abilities than conventional human ESC and hiPSC including more rapid expansion in culture, greater differentiation potencies, and a higher capacity to be genetically manipulated by gene targeting with plasmid-based homologous recombination. We will generate and differentiate N-hiPSC, and specifically test for their capacity to undergo gene targeting and to produce functional embryonic VP that functionally integrate into ischemic tissues for future cell therapies. Finally, we will conduct detailed comparative epigenetic analyses of na�ve and conventional hiPSC and their differentiated VP progeny to elucidate differential CpG methylation and histone architectures. We propose that patient-specific na�ve hiPSC will ultimately provide greater versatility for cellular and gene therapies for pediatric and adult regenerative medicine.