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

描述(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。这些情况包括老年性黄斑变性和视网膜色素变性，其特征是视网膜外层细胞进行性丧失。干细胞通过使丢失的细胞重新繁殖，为治疗这些疾病带来了巨大的希望。视网膜色素上皮(RPE)是这一策略成功的核心细胞类型。最近的技术突破首次使通过重新编程成人细胞的多能性和定向分化来生产受者特有的捐赠者细胞成为可能。然而，目前尚不清楚这些方法和其他方法产生的RPE细胞在关键生物学特性方面的比较，包括免疫原性和线粒体衰老，当移植到健康或患病的视网膜时。这些问题对于将这种细胞用于视网膜疾病治疗的潜在用途至关重要。该方案的目的是研究由实验诱导的多能干细胞在体外以及移植到啮齿动物或非人灵长类视网膜后所产生的RPE细胞的功能，包括它们的免疫原性以及它们在视网膜变性的啮齿动物模型中挽救视力丧失的能力。该项目将创新地利用独特的资源，包括同种异体移植和自体移植干细胞来源的恒河猴RPE细胞系，以及自然发生的非人类灵长类黄斑疾病模型。该建议有三个具体目标：1)从三种来源的多潜能干细胞--胚胎干细胞(ESCs)、体细胞核移植来源的ESCs和诱导的多潜能干细胞(IPS)--培养恒河猴RPE细胞，并评估它们在阻止RCS视网膜变性模型中视力衰退方面的功能。2)研究这些RPE细胞作为同种异体或自体RPE细胞移植到成年恒河猴视网膜及同眼或对侧眼再治疗后的免疫学特性。3)研究幼龄恒河猴和老年恒河猴免疫反应的差异，包括与人类AMD类似的老年性黄斑病变。这一转化研究项目将为视网膜细胞治疗的成功提供关键信息。它将提供关于最合适的细胞来源的见解，以重新填充丢失的视网膜细胞，以保护或恢复视力，并将产生关于此类治疗干预的免疫反应的新数据，以及避免移植物排斥反应的最佳方法。该提案将在一种动物模型中解决这些问题，该动物模型的眼睛和免疫系统最接近人类，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.