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

基本信息

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
项目类别：
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

项目摘要

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能是。用自体或细胞库自我更新血管周细胞干祖细胞再生，缺血我们组的这些血管病可以得到缓解，并且终末期失明得到逆转或稳定。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我们还建立了一种基于坦科聚合酶/PARP 抑制剂的新型小分子混合物。传统的、谱系引发的 hiPSC 恢复为“原始”hiPSC（N-hiPSC），其具有更多原始外胚层状态，具有较高的功能多能性。幼稚 VP (N-VP) 的再生潜力。与引发的、例如，原始糖尿病血管祖细胞（N-DVP）分化为传统的 hiPSC。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，其效率更高在本提案中，我们从传统的、引发的 DhiPSC 中获得同基因糖尿病血管祖细胞 (DVP)。开发 N-VP 治疗缺血性视网膜病的潜力，我们将采用人源化药物。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 形成通畅血管、挽救缺血性视网膜、改善视力的治疗能力我们将测试 N-DVP 的体内发育潜力，以有效分化为长期植入缺血损伤后的内皮细胞和多能周细胞干祖细胞我们还将致力于进一步改进我们产生无限量的方法。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。研究将勾勒出一条未来有效同步生成幼稚血管和来自同一 N-hiPSC 系的视网膜干祖细胞，以获得更有效和更全面的更广泛地说，我们将开发这类新型药物的临床前用途。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。