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: 10334409
• 负责人: ELIAS T. ZAMBIDIS
• 金额: $ 53.48万
• 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/10334409
• 关键词: Adult; Animal Model; Autologous; Back; Blindness; Blood Vessels; Blood capillaries; CXCR4 gene; Cell Line; Cell Therapy; Cells; Cessation of life; DNA Methylation; Defect; Development; Diabetes Mellitus; Diabetic Retinopathy; Disease; Embryo; Endothelial Cells; Engraftment; Epiblast; Epigenetic Process; Eye; Fibroblasts; 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; PECAM1 gene; Pathologic; Pathology; Pathway interactions; Patients; Pericytes; Regenerative Medicine; Reperfusion Injury; Reperfusion Therapy; Reporting; Retina; Retinal Diseases; Secondary to; Tankyrase; Therapeutic; Tissue Differentiation; Tissues; Transplant Recipients; Transplantation; Vascular Diseases; Vascular Endothelial Cell; Vision; base; cell bank; diabetic; effectiveness evaluation; endothelial stem cell; epigenetic memory; epigenome; genome integrity; improved; improved functioning; in vivo; in vivo evaluation; induced pluripotent stem cell; inhibitor; mouse model; neuron apoptosis; neurovascular injury; novel; pluripotency; pre-clinical; progenitor; promoter; regeneration function; regeneration potential; relating to nervous system; repaired; retina blood vessel structure; retinal damage; retinal ischemia; self-renewal; small molecule; stem; 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）和糖尿病视网膜病变（DR）是新发失明的主要原因 美国。这些血管疾病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（EC）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血管周细胞干祖细胞再生，缺血 这些血管病变的症状可以得到缓解，并且终末期失明得到逆转或稳定。我们组 建立了移植患者特异性胚胎血管祖细胞（VP）的可行性 从人类诱导多能干细胞分化后，周细胞电位直接进入眼睛 细胞（hiPSC）。我们还建立了一种基于端锚聚合酶/PARP 抑制剂的新型小分子混合物 将传统的、谱系引发的 hiPSC 逆转为“原始”hiPSC（N-hiPSC），该 hiPSC 具有更多 具有较高功能多能性的原始外胚层状态。幼稚 VP 的再生潜力 (N-VP) 从正常和患病的 N-hiPSC 中分化出来的 N-hiPSC 相对于引发的 N-hiPSC 的繁殖力显着更高， 传统 hiPSC。例如，幼稚糖尿病血管祖细胞（N-DVP）分化为 患者特异性的幼稚回复型糖尿病 hiPSC (N-DhiPSC) 具有更高的血管功能， 保持更高的基因组稳定性，降低谱系引发的基因表达，并且 与缺血性视网膜深层神经层的迁移和血管重建相比，它更有效 来自传统、引发的 DhiPSC 的同基因糖尿病血管祖细胞 (DVP)。在这个提案中，我们 开发 N-VP 治疗缺血性视网膜病的潜力。我们将采用人性化的 模拟视网膜缺血[即缺血/再灌注（I/R）损伤]的动物模型，用于测试 人 N-DVP 形成开放血管、挽救缺血性视网膜、改善视力的治疗能力 视觉功能。我们将测试 N-DVP 的体内发育潜力，以有效分化 长期植入缺血损伤后的内皮细胞和多能周细胞干祖细胞 视网膜生态位。我们还将致力于进一步改进我们产生无限数量的方法 表观遗传可塑性、原始、无病的幼稚胚胎祖细胞，用于细胞治疗 探究 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 hiPSC 重编程相比，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生成天然血管和 来自同一 N-hiPSC 系的视网膜干祖细胞，以获得更有效和更全面的 患病视网膜的再生。更广泛地说，我们将开发这类新型药物的临床前效用 人类血管周细胞干细胞具有高表观遗传可塑性、改进的功能和 对眼再生医学具有潜在的巨大影响。