Pluripotent Stem Cell Vascular Therapies for Ischemic Retinopathies

描述(申请人提供)：分支静脉阻塞(BVO)和糖尿病视网膜病变(DR)是美国新发失明的主要原因。由于视网膜血管内皮细胞(ECs)和收缩周细胞的缺血性死亡，这两种疾病都会导致无细胞的毛细血管。脱细胞视网膜毛细血管能否用自体血管/周细胞重新填充 这些疾病可缓解先祖细胞的缺血，逆转终末期失明。一种这样的方法是移植患者特定的胚胎血管前体细胞(VP)，具有丰富的血管和间充质-周细胞潜能。这样的祖细胞可以从人类诱导的多能干细胞(HiPSC)中产生，并直接移植到眼睛中。这种方法可以与来自同一个HiPSC系的平行分化相结合，以取代缺血变性的视网膜神经组织及其所需的血管壁龛。到目前为止，还没有人评估HiPSC来源的胚胎VP对移植和挽救缺血视网膜中退化的血管形成的能力。在这个方案中，我们将测试HiPSC来源的胚胎VP在植入受损的缺血视网膜后有效分化为内皮细胞和周细胞的潜力。我们将使用模拟人类BVO[即缺血/再灌注(I/R)损伤]或DR(在无菌大鼠中诱导)的动物模型来测试HiPSC衍生的VP形成通畅血管和拯救缺血视网膜的潜力。我们将把表达内皮(CD31+)和周细胞-间充质(CD146+)标记的HiPSC来源的CXCR4+VP直接注射到免疫缺陷NOD/SCID小鼠的玻璃体间隙(或IV眶窦)，这些小鼠的眼睛已经被实验性地因I/R损伤或糖尿病而退化。然后，我们将测试这些祖细胞的能力，以了解它们通过毛细血管重新填充和再生、挽救神经视网膜和改善视觉功能的能力。我们还将确定视网膜微环境损伤和低氧相关信号，这些信号调节CXCR4+胚胎VP归巢和植入无细胞视网膜毛细血管。我们推测，从非病毒髓系细胞高效生成血管前体细胞最终将在治疗缺血性眼病方面具有更好的临床应用价值。如果成功，我们的方法将允许容易地从患者自己的血细胞中重新编程产生自体、多潜能和临床有用的血管形成前体细胞，用于BVO和DR的临床治疗。

DESCRIPTION (provided by applicant): Branch vein occlusion (BVO) and diabetic retinopathy (DR) are the major causes of new onset blindness in the US. Both disorders result in acellular capillaries due to ischemic death of retinal vascular endothelial cells (ECs) and contractile pericytes. If acellular retinal capillaries could be repopulated with autologous vascular/pericytic progenitors, ischemia could be relieved and end stage blindness reversed in these diseases. One such approach is to transplant patient-specific embryonic vascular progenitors (VP) with prolific vascular and mesenchymal-pericytic potential. Such progenitors could be generated from human induced pluripotent stem cells (hiPSC) and transplanted directly into the eye. This approach could be combined with parallel differentiation from the same hiPSC line to replace ischemic degenerated retinal neural tissue along with their requisite vascular niche. To date, no one has evaluated hiPSC-derived embryonic VP for capacity to engraft and rescue degenerated vasculature in the ischemic retina. In this proposal, we will test the potential of hiPSC-derived embryonic VP to efficiently differentiate to ECs and pericytes following engraftment into damaged ischemic retina. We will use animal models that mimic human BVO [i.e., ischemia/reperfusion (I/R) injury] or DR (induced in athymic rats) for testing the potential of hiPSC-derived VP to form patent blood vessels and rescue ischemic retina. We will inject hiPSC-derived CXCR4+ VP expressing endothelial (CD31+) and pericytic- mesenchymal (CD146+) markers directly into the vitreous space (or IV orbital sinus) of immunodeficient NOD/scid mouse eyes that have been experimentally degenerated by I/R injury or diabetes. We will then test the ability of these progenitors for their ability to repopulate and regenerate viabe capillaries, rescue neural retina, and improve visual function. We will also determine the retinal micro-environmental injury and hypoxia-related signals that regulate homing and engraftment of CXCR4+ embryonic VPs to acellular retinal capillaries. We hypothesize that efficient generation of vascular progenitors from nonviral myeloid-iPSC will ultimately have superior clinical utility fr the treatment of ischemic ocular diseases. If successful, our approach would allow facile generation of autologous, multipotent, and clinically useful vascular-forming precursors reprogrammed from a patient's own blood cells to be used in clinical therapies for BVO and DR.