Retinal disease models for translational photoreceptor replacement

用于平移感光器替代的视网膜疾病模型

• 批准号: 10063767
• 负责人: William A. Beltran
• 金额: $ 21.36万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063767
• 关键词: 3-Dimensional; Animal Model; Behavioral Assay; Biological; Blindness; Brain; CNS processing; Canis familiaris; Cell Count; Cell Differentiation process; Cell Survival; Cell Transplantation; Cells; Cessation of life; Clinical Trials; Disease; Disease model; Dog Diseases; Engraftment; Eye diseases; Foundations; Functional Magnetic Resonance Imaging; Future; Gene therapy trial; Generations; Genes; Goals; Homologous Gene; Human; Hydrogels; Immunosuppression; Impairment; Inherited; Injections; Knowledge; Label; Lead; Leber' s amaurosis; Measures; Mediating; Methods; Modeling; Mutation; Natural regeneration; Neural Retina; Orthologous Gene; Outcome; Outcome Measure; Pathologic; Pathology; Patients; Phase I/II Clinical Trial; Photoreceptors; Play; Pluripotent Stem Cells; Property; Protocols documentation; Psychophysics; RPE65 protein; Recovery of Function; Regenerative Medicine; Replacement Therapy; Reporter; Research; Research Personnel; Retina; Retinal Degeneration; Retinal Diseases; Retinal Photoreceptors; Retinal Pigments; Retinal gene therapy; Rod; Role; Safety; Site; Specific qualifier value; Structure; Structure of retinal pigment epithelium; System; Therapeutic; Transplantation; Vertebrate Photoreceptors; Vision; Visual; Visual Cortex; Visual Pathways; Xenograft procedure; base; behavior test; blind; clinically relevant; cost effective; experimental analysis; gene therapy; human disease; human model; human pluripotent stem cell; improved; induced pluripotent stem cell; inherited retinal degeneration; multi-electrode arrays; nerve stem cell; precursor cell; prevent; relating to nervous system; research facility; response; restoration; retina transplantation; safety study; scaffold; stem cell biology; stem cells; success; translational model; translational study; treatment response

Diseases of the neural retina or retinal pigment epithelium (RPE) cause a substantial number of sight-impairing disorders, many of which lead to the degeneration and death of photoreceptor cells. A number of naturally occurring inherited retinal degeneration (RD) diseases have been identified in dogs that are true homologues of human diseases. Our research team has been instrumental in demonstrating that AAV-mediated gene therapy for the retina in canine disease models plays a critical role in the translational continuum by permitting a rapid, cost-effective, and clinically relevant assessment of therapeutic responses and long-term outcomes of retinal gene therapy. However, the success of prior gene therapy trials depended on the presence of viable, albeit diseased, target cells. In contrast, the substantial loss of target cells in advanced RD will require therapeutic strategies that permit regeneration or replacement of photoreceptor cells and restoration of neural connectivity. Dog models of human retinal diseases can provide the foundational knowledge needed for photoreceptor replacement therapies to improve visual function, as specified in the AGI RFA. We have assembled a team of investigators with expertise in neural progenitor and pluripotent stem cell biology, retinal cell differentiation, inherited retinal disease pathology and therapy, and functional analysis of experimental RD treatments. The investigators have the expertise to assess therapeutic responses and to develop appropriate outcome measures to evaluate safety and efficacy. We have a dedicated retinal disease research facility to breed, maintain, treat and assess dogs with homologous human retinal disorders. Our goal is to develop canine models in which to investigate the replacement of photoreceptors under disease conditions. The dog models will be used to identify key parameters of cell transplantation, engraftment, and differentiation that will be critical for studies of disease-specific applications of regenerative medicine for the retina. Key properties to be studied are physical transplantation parameters, distribution of donor cells within the host retina, donor cell differentiation and survival, cell connectivity and functionality within the retina, connectivity to the visual pathways in the brain and CNS processing, and behavioral assays for vision. Transplantation parameters will be studied initially using xenografts derived from two well-characterized and readily available human pluripotent stem cell reporter lines that label cones and rods or only rods. Concurrently, we will develop a within-species system for photoreceptor replacement by producing equivalent canine iPSCs and cone/rod and rod-only iPSC reporter lines for replacement of retinal photoreceptor cells to recapitulate the engraftment that would occur in human clinical trials using human cells. Once developed, the canine photoreceptor precursor cell transplants will be analyzed for integration and functionality in diseased dog models in which differentiation into new rod and cone visual cells can be evaluated.

神经视网膜或视网膜色素上皮（RPE）的疾病引起大量观察 疾病，其中许多导致感光细胞的变性和死亡。许多自然 发生的遗传性视网膜变性（RD）疾病已在真正的同源物中发现 人类疾病。我们的研究团队在证明AAV介导的基因方面发挥了作用 犬类病模型中视网膜的治疗在翻译连续体中起着至关重要的作用 对治疗反应和长期结局的快速，成本效益和临床相关评估 视网膜基因疗法。但是，先前基因治疗试验的成功取决于存在的存在， 尽管患病是靶细胞。相反，高级RD中目标细胞的大量损失将需要 允许再生或替代感光细胞并恢复神经的治疗策略 连接性。人类视网膜疾病的狗模型可以提供所需的基本知识 如AGI RFA所示，光感受器替代疗法可改善视觉功能。我们有 组建了一个研究人员团队，具有神经祖细胞和多能干细胞生物学，视网膜的专业知识 细胞分化，遗传性视网膜疾病病理和治疗以及实验RD的功能分析 治疗。调查人员具有评估治疗反应并发展适当的专业知识 结果措施评估安全性和有效性。我们有专门的视网膜疾病研究机构 繁殖，维持，治疗和评估人类视网膜疾病的狗。 我们的目标是开发犬类模型，以调查替换感光体的替代 疾病状况。狗模型将用于识别细胞移植的关键参数， 植入和分化对于疾病特异性应用至关重要 视网膜的药物。要研究的关键属性是物理移植参数，分布 宿主视网膜内的供体细胞，供体细胞分化和存活，细胞连接性和功能 视网膜，与大脑和中枢神经系统处理的视觉途径的连通性以及行为分析 想象。最初，使用源自两个良好特征的异种移植物来研究移植参数 并随时可用的人多能干细胞报告剂线，标记锥和杆或仅标记杆。 同时，我们将通过产生等价的物种内物种系统来替换感光器 犬IPSC和锥/杆和仅杆IPSC报告基因线，用于将视网膜感光细胞替换为 概括使用人类细胞在人类临床试验中发生的植入。一旦开发， 将分析犬类感受器前体细胞移植物的整合和功能 可以评估将分化为新棒和锥体视觉细胞的狗模型。