Evaluation of stem cell-derived retinal pigment epithelial cells for retinal dise

描述（由申请人提供）：视网膜退行性疾病累积起来是导致无法治愈的失明的最常见原因。这些疾病包括年龄相关性黄斑变性和视网膜色素变性，其特征是外层视网膜细胞的进行性丧失。干细胞通过重新填充已经丢失的细胞，在治疗这些疾病方面具有很大的希望。视网膜色素上皮细胞（RPE）是这种策略成功的核心细胞类型。最近的技术突破使得首次通过在成人细胞中重新编程多能性和定向分化来生产受体特异性供体细胞成为可能。然而，目前尚不清楚通过这些方法和其他方法产生的RPE细胞在移植到健康或病变视网膜时，在关键生物学特性（包括免疫原性和线粒体衰老）方面的比较。这些问题对于这些细胞在视网膜疾病治疗中的潜在应用至关重要。本研究的目的是研究实验诱导的多能干细胞在体外和移植到啮齿动物或非人灵长类动物视网膜后产生的RPE细胞的功能，包括它们的免疫原性和它们在啮齿动物视网膜变性模型中恢复视力丧失的能力。该项目将创新性地利用独特的资源，包括异体移植和自体移植干细胞衍生的恒河猴RPE细胞系和自然发生的非人类黄斑疾病灵长类动物模型。该提案有三个具体目标：1)从三种多能干细胞来源——胚胎干细胞（ESCs）、体细胞核移植（SCNT- ESCs）衍生的ESCs和诱导多能干细胞（iPS）细胞中生成恒河猴RPE细胞，并评估它们在RCS视网膜变性模型中阻止视力下降的功能。2)观察同种异体或自体RPE细胞移植到成年恒河猴视网膜，并在同眼或同眼再治疗后的免疫学情况。3)研究年轻恒河猴和衰老恒河猴（包括与年龄相关的黄斑病变类似的中度人类AMD）免疫反应的差异。这个转化研究项目将为视网膜细胞治疗的成功提供关键信息。它将提供最合适的细胞来源来重新填充丢失的视网膜细胞，以保持或恢复视力，并将产生对这种治疗干预的免疫反应的新数据和避免移植排斥的最佳方法。该提案将在具有与人类最相似的眼睛和免疫系统的动物模型中解决这些问题，并以一种密切反映潜在临床实践的方式。

DESCRIPTION (provided by applicant): Degenerative diseases of the retina are cumulatively the most common causes of untreatable blindness. These conditions, which include age-related macular degeneration and retinitis pigmentosa, are characterized by progressive loss of cells in the outer retina. Stem cells hold great promise for treating these diseases by repopulating cells that have been lost. A cell type that will be central to the success of this strategy is the retinal pigment epithelium (RPE). Recent technological breakthroughs make possible for the first time the production of recipient-specific donor cells through reprogramming of pluripotency in adult cells and directed differentiation. However, it is not known how RPE cells produced by these and other methods compare with respect to key biological characteristics, including immunogenicity and mitochondrial senescence, when transplanted to the healthy or diseased retina. These issues are critical to the potential use of such cells for retinal disease therapy. The goal of this proposal is to study the functionality of RPE cells generated from experimentally induced pluripotent stem cells in vitro and after transplantation to the rodent or nonhuman primate retina, including their immunogenicity and their ability to rescue visual loss in a rodent model of retinal degeneration. The project will make innovative use of unique resources, including allograft and autograft stem-cell-derived rhesus monkey RPE cell lines and a naturally-occurring nonhuman primate model of macular disease. The proposal has three specific aims: 1) To generate rhesus macaque RPE cells from three sources of pluripotent stem cells--embryonic stem cells (ESCs), ESCs derived by somatic cell nuclear transfer (SCNT- ESCs), and induced pluripotent stem (iPS) cells--and evaluate their function in arresting visual decline in the RCS retinal degeneration model. 2) To investigate the immunology of these RPE cell types transplanted as allograft or autografts into the retina of adult rhesus monkeys and after retreatment of the same eye or fellow eye. 3) To characterize the differences in immune response between young and senescent rhesus monkeys, including those with age-related maculopathy which parallels intermediate human AMD. This translational research project will provide information key to the success of cell therapy in the retina. It will provide insights as to the most appropriate cell source to repopulate lost retinal cells with the aim of preserving or restoring vision, and will generate novel data on the immune response to such therapeutic intervention and the best approach to avoid graft rejection. The proposal will address these issues in an animal model with an eye and immune system that most closely resembles that of humans and in a manner closely mirroring potential clinical practice.