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: 10006534
• 负责人: William A. Beltran
• 金额: $ 137.35万
• 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/10006534
• 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; 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; photoreceptor degeneration; 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 that are true homologues of human diseases. Our research team has been instrumental in demonstrating that AAV-mediated gene therapy for the retina in 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. 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 assess models of human retinal disorders. Our goal is to develop models in which to investigate the replacement of photoreceptors under disease conditions. The 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 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 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 photoreceptor precursor cell transplants will be analyzed for integration and functionality in disease 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) 疾病会导致大量视力损害疾病，其中许多会导致感光细胞变性和死亡。一些自然发生的遗传性视网膜变性（RD）疾病已被鉴定为人类疾病的真正同源物。我们的研究团队在证明疾病模型中 AAV 介导的视网膜基因治疗在转化连续体中发挥着关键作用，通过允许对治疗反应和长期结果进行快速、具有成本效益和临床相关的评估，发挥了重要作用。视网膜基因治疗。然而，先前基因治疗试验的成功取决于存活的（尽管患病的）靶细胞的存在。相比之下，晚期 RD 中靶细胞的大量损失将需要允许感光细胞再生或替换以及神经连接恢复的治疗策略。根据 AGI RFA 的规定，人类视网膜疾病模型可以提供光感受器替代疗法所需的基础知识，以改善视觉功能。我们组建了一支研究人员团队，他们在神经祖细胞和多能干细胞生物学、视网膜细胞分化、遗传性视网膜疾病病理学和治疗以及实验性 RD 治疗的功能分析方面拥有专业知识。研究人员拥有评估治疗反应和制定适当的结果措施来评估安全性和有效性的专业知识。我们拥有专门的视网膜疾病研究设施来评估人类视网膜疾病模型。 我们的目标是开发模型来研究疾病条件下光感受器的替换。这些模型将用于确定细胞移植、植入和分化的关键参数，这对于研究视网膜再生医学的特定疾病应用至关重要。要研究的关键特性包括物理移植参数、供体细胞在宿主视网膜内的分布、供体细胞分化和存活、视网膜内的细胞连接性和功能、与大脑和中枢神经系统处理中视觉通路的连接性以及视觉行为分析。将使用源自两种特征良好且易于获得的人类多能干细胞报告系的异种移植物来研究移植参数，这些报告系标记视锥细胞和视杆细胞或仅标记视杆细胞。同时，我们将开发一种用于光感受器替代的种内系统，通过生产等效的 iPSC 和视锥/视杆和仅视杆 iPSC 报告系来替代视网膜感光细胞，以重现使用人类细胞的人类临床试验中发生的植入。一旦开发完成，将分析光感受器前体细胞移植物在疾病模型中的整合和功能，在疾病模型中可以评估分化为新的视杆细胞和视锥细胞。