Embryonic vascular stem-progenitors for treatment of ischemic retinopathies

用于治疗缺血性视网膜病的胚胎血管干祖细胞

• 批准号: 10557078
• 负责人: ELIAS T. ZAMBIDIS
• 金额: $ 52.92万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10557078
• 关键词: Adult; Animal Model; Autologous; Back; Blindness; Blood Vessels; Blood capillaries; CXCR4 gene; Cell Line; Cell Reprogramming; Cell Therapy; Cells; Cessation of life; DNA Methylation; Defect; Development; Diabetes Mellitus; Diabetic Retinopathy; Disease; Embryo; Embryo Transfer; Endothelial Cells; Engraftment; Epiblast; Epigenetic Process; Eye; Fibroblasts; Functional Regeneration; Future; Gene Expression; Generations; Genome Stability; Histones; Human; In Vitro; Ischemia; Legal patent; Lesion; MCAM gene; Mediating; Memory; Metabolic; Modeling; Molecular; Mus; Natural regeneration; Neural Retina; PARP inhibition; PECAM1 gene; Pathologic; Pathology; Pathway interactions; Patients; Pericytes; Poly(ADP-ribose) Polymerase Inhibitor; Regenerative Medicine; Reperfusion Injury; Reperfusion Therapy; Reporting; Retina; Retinal Diseases; Secondary to; Tankyrase; Therapeutic; Tissue Differentiation; Tissues; Transplant Recipients; Vascular Diseases; Vascular Endothelial Cell; Vascularization; Vision; cell bank; derepression; diabetic; effectiveness evaluation; endothelial stem cell; epigenetic memory; epigenome; functional improvement; genome integrity; improved; in vivo; in vivo engraftment; in vivo evaluation; induced pluripotent stem cell; migration; mouse model; neural; neuron apoptosis; neurovascular injury; novel; pluripotency; pre-clinical; progenitor; promoter; regeneration function; regeneration potential; repaired; retina blood vessel structure; retinal damage; retinal ischemia; self-renewal; small molecule; stem; stem cell expansion; stem cell self renewal; stem cells; therapeutic evaluation; type I diabetic; vein occlusion

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.

分支静脉阻塞（BVO）和糖尿病性视网膜病（DR）是新发作失明的主要原因 美国。这些血管疾病导致视网膜缺血性死亡继发的细胞毛细血管 血管内皮细胞（ECS）和收缩周周细胞。如果可以是巨细胞残留的血管 用自体或细胞储备的自我更新血管 - 磨性干核病患者的再生，缺血 我们可以放心，在这些血管疾病中，终结阶段的失明逆转或稳定。我们的小组 建立了使用患者特异性胚胎血管祖细胞（VP）的可行性 与人类诱导的多能茎分化后，直接进入眼睛的细胞电位直接进入眼睛 细胞（HIPSC）。我们还建立了一种新型的罐式酶/PARP抑制剂的小分子鸡尾酒 传统的谱系hipscs恢复为“幼稚” hipscs（n-ipscs），该hipscs拥有更多 具有较高功能多能性的原始层细胞状态。幼稚VP（N-VP）的再生潜力 相对于Primed，与正常和解散的N-Hipsc区分开明显更加多产 常规HIPSC。例如，幼稚的糖尿病血管祖细胞（N-DVP）与 患者特异性的幼稚糖尿病hipsc（N-Dhipsc）具有较高的血管功能， 保持较高的基因组稳定性，含有谱系引发基因表达降低，并且 比迁移和重新血管更有效地使缺血性视网膜的深神经层次更高 来自常规的Priment dhipsc的等源性糖尿病血管祖细胞（DVP）。在这个建议中，我们 开发N-VP治疗缺血性视网膜病的潜力。我们将雇用人生的 模仿视网膜缺血的动物模型[即缺血/再灌注（I/R）损伤] 人类N-DVP的治疗能力形成专利血管，营救缺血性视网膜并改善 视觉功能。我们将测试N-DVP的体内发育潜力，以有效区分 长期植入后，ECS和多能细胞性干螺旋体在缺血损害中 视网膜利基。我们还将旨在进一步改善我们生成无限量的方法 表观遗传塑料，原始的，未固定的幼稚胚胎祖细胞，用于细胞疗法 探测n-hipsc重新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 - 与常规HIPSC重编程相比，相关的代谢畸变具有更高的效率。这些 研究将概述有效同步生成幼稚的血管和 来自同一N-HIPSC线的视网膜干螺旋体，以更有效，更全面 解散视网膜的再生。更广泛地说，我们将发展这一新颖的类别的临床前实用性 具有高表观遗传可塑性，改善功能和的人血管生存干细胞 对眼部再生医学的潜在高影响。