Embryonic vascular stem-progenitors for treatment of ischemic retinopathies

分支静脉阻塞(BVO)和糖尿病视网膜病变(DR)是新发致盲的主要原因 美国。这些血管紊乱导致继发于视网膜缺血性死亡的无细胞毛细血管。 血管内皮细胞(ECs)和收缩周细胞。如果无细胞视网膜血管可以 用自体或细胞库自我更新的血管周细胞干细胞再生，缺血 在这些血管病变中可以缓解和逆转或稳定终末期失明。我们的团队 建立了患者特异性胚胎血管祖细胞移植的可行性 人诱导多能干细胞分化后，周细胞潜能直接进入眼睛 细胞(HiPSC)。我们还建立了一种新型的基于tankyrase/PARP抑制剂的小分子鸡尾酒。 将传统的、谱系启动的HiPSC回复到具有更多 原始的上胚层状态，具有较高的功能多能性。朴素VP(N-VP)的再生潜力 与正常和患病的N-hiPSC相比，分化的N-hiPSC显著更多。 传统的HiPSC。例如，幼稚的糖尿病血管祖细胞(N-DVP)与 患者特异性天真逆转的糖尿病HiPSC(N-DhiPSC)具有更高的血管功能， 保持了更大的基因组稳定性，抑制了谱系启动的基因表达减少，并 更有效地迁移到缺血视网膜的深层神经层并使其重新血运 同基因糖尿病血管祖细胞(DVP)来自常规的，预充的DhiPSC。在这项提案中，我们 开发N-VP治疗缺血性视网膜病变的潜力。我们将雇佣一个人性化的 模拟视网膜缺血[即缺血/再灌注(I/R)损伤]的动物模型 人N-DVP形成通畅血管、挽救缺血视网膜和改善视网膜功能的治疗能力 视觉功能。我们将测试N-DVP的体内发育潜力，以有效地分化为 缺血损伤后长期植入内皮细胞和多潜能周细胞干祖细胞 视网膜小窝。我们还将致力于进一步改进我们的方法，以产生无限数量的 表观遗传学-用于细胞治疗的可塑的、原始的、未患病的幼稚胚胎祖细胞 探索N-hiPSC重编程如何消除功能失调的表观遗传供体细胞记忆和糖尿病- 与传统的HiPSC重新编程相比，相关代谢异常的效率更高。这些 研究将勾勒出一条有效同步产生幼稚维管束和 来自同一N-hiPSC系的视网膜干祖细胞更有效和更全面 病变视网膜的再生。更广泛地说，我们将开发这类新的 人类血管周细胞干细胞具有高度的表观遗传可塑性，改善的功能，以及 可能对眼部再生医学产生很大影响。

Branch vein occlusion (BVO) and diabetic retinopathy (DR) are major causes of new onset blindness in the US. These vascular disorders result in acellular capillaries secondary to ischemic death of retinal vascular endothelial cells (ECs) and contractile pericytes. If acellular retinal blood vessels could be regenerated with autologous or cell-banked self-renewing vascular-pericytic stem-progenitors, ischemia could be relieved, and end stage blindness reversed or stabilized in these vasculopathies. Our group established the feasibility of transplanting patient-specific embryonic vascular progenitors (VP) with pericytic potential directly into the eye, following differentiation from human induced pluripotent stem cells (hiPSC). We also established a novel tankyrase/PARP inhibitor-based small molecule cocktail for reversion of conventional, lineage-primed hiPSC to ‘naïve’ hiPSCs (N-hiPSCs) that possessed a more primitive epiblast state with higher functional pluripotency. The regenerative potential of naïve VP (N-VP) differentiated from normal and diseased N-hiPSC was significantly more prolific relative to primed, conventional hiPSC. For example, naive diabetic vascular progenitors (N-DVP) differentiated from patient-specific naïve-reverted diabetic hiPSC (N-DhiPSC) possessed higher vascular functionality, maintained greater genomic stability, harbored decreased lineage-primed gene expression, and were more efficient in migrating to and re-vascularizing the deep neural layers of the ischemic retina than isogenic diabetic vascular progenitors (DVP) from conventional, primed DhiPSC. In this proposal, we develop the potential of N-VP for treatment of ischemic retinopathies. We will employ a humanized animal model that mimics retinal ischemia [i.e., ischemia/reperfusion (I/R) injury] for testing the therapeutic capacity of human N-DVP to form patent blood vessels, rescue ischemic retina, and improve visual function. We will test the in vivo developmental potential of N-DVP to efficiently differentiate to ECs and multipotent pericytic stem-progenitors following long-term engraftment in an ischemia-damaged retinal niche. We will also aim to further improve our approach for generating unlimited amounts of epigenetically-plastic, pristine, non-diseased naïve embryonic progenitors for cellular therapies by probing how N-hiPSC reprogramming erases dysfunctional epigenetic donor cell memory and diabetes- associated metabolic aberrations with greater efficiency than conventional hiPSC reprogramming. These studies will outline a future pathway for the efficient synchronous generation of naïve vascular and retinal stem-progenitors from the same N-hiPSC line for a more effective and comprehensive regeneration of diseased retina. More broadly, we will develop the pre-clinical utility of this novel class of human vascular-pericytic stem cells that possess high epigenetic plasticity, improved functionality, and potentially high impact for ocular regenerative medicine.