Developing Cone-Dominant Retinal Disease Models as a Resource for Translational Vision Research

开发视锥细胞为主的视网膜疾病模型作为转化视觉研究的资源

• 批准号: 10477216
• 负责人: Joseph Carroll
• 金额: $ 123.89万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 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/10477216
• 关键词: Affect; Affinity; Alleles; Anatomy; Animal Model; Animals; Area; BCL9 gene; Biological Assay; Biological Models; Brain region; Cell Survival; Cells; Chemicals; Collaborations; Color; Communities; Cone; Data; Development; Discrimination; Disease; Disease model; Dominant-Negative Mutation; Embryo; Evaluation; Eye; Functional Magnetic Resonance Imaging; Functional disorder; Gene Mutation; Genes; Genome; Goals; Hibernation; Human; Image; In Vitro; Individual; Logistics; Mammals; Mediating; Methods; Micromanipulation; Modeling; Molecular Genetics; Morphogenesis; Mosaicism; Mus; Mutation; Myopia; Natural regeneration; Neurons; Organoids; Patients; Phenotype; Photoreceptors; Phototransduction; Pluripotent Stem Cells; Primates; Process; Protocols documentation; Rattus; Recombinant adeno-associated virus (rAAV); Recording of previous events; Resolution; Resources; Retina; Retinal Cone; Retinal Degeneration; Retinal Detachment; Retinal Diseases; Rod; Rodent; Rodent Model; Scandentia; Signal Transduction; Spermophilus; Stem Cell Research; Structure; Study models; Techniques; Technology; Testing; Therapeutic; Transgenic Organisms; Translations; Transplantation; Tupaiidae; Validation; Vision; Vision research; Visual; Visual Cortex; Visual system structure; Work; adaptive optics scanning laser ophthalmoscopy; area striata; cone-rod dystrophy; density; disease phenotype; fovea centralis; genetic manipulation; human disease; human model; imaging modality; in vivo; induced pluripotent stem cell; innovation; interest; model development; monolayer; multidisciplinary; nonhuman primate; novel therapeutics; overexpression; regenerative therapy; regenerative treatment; relating to nervous system; reproductive; retinal imaging; stem cell therapy; tool; tool development; treatment strategy; two-photon; visual information; visual processing

PROJECT SUMMARY/ABSTRACT . The NEI’s Audacious Goal Initiative (launched in 2012) put forward the challenge of “restoring usable vision in humans by regenerating neurons and neural connections in the eye and visual system.” While there is an obvious affinity towards novel therapies, current resource and technology gaps preclude translation of many therapeutic approaches. One such gap pertains to the availability of animal models that share key features of human retinal anatomy, as well as disease models that faithfully emulate the mechanisms and processes seen in patients with retinal degenerations (blinding diseases that might be amenable to regenerative therapies). The absence of readily available cone-dominant mammalian models represents a major technology gap impeding efforts to develop and evaluate regenerative treatment strategies in the retina. We propose to advance two promising model systems that are closer to human visual anatomy and function than the more widely used mouse and rat models. The first is the 13-lined ground squirrel (13-LGS): a diurnal, cone-dominant rodent (~85% cones) with large brain regions dedicated to processing visual information. The second is the tree shrew: a non- rodent, primate-like mammal that is also cone dominant (~95% cones). These models have been used to study visual transduction (13-LGS), outer segment morphogenesis, shedding, and remodeling during hibernation (13- LGS), cone-bipolar cell circuitry (13-LGS), myopia (tree shrew) and central visual processing (tree shrew). However, their use as translation-enabling models for evaluating both survival and integration of regenerated cone photoreceptors has been limited; mainly due to a lack of tools that allow for genetic manipulation of these animals (and thus a dearth of disease models). We propose to advance these species as disease-relevant models through the following Specific Aims: (1) Develop, optimize, and validate imaging methods and functional assays for the 13-LGS and tree shrew; (2) Generate 13-LGS and tree shrew cone photoreceptors from iPSCs in vitro; (3) Create rAAV-mediated retinal degeneration models for the 13-LGS and tree shrew in vivo; (4) Enable germline transgenic 13-LGS models of human disease; (5) Test and optimize integration of transplanted 13- LGS, tree shrew, and human iPSC-derived cones in normal and degenerated 13-LGS and tree shrew retinas. A key feature of this proposal is the validation of these models by comparing their cellular-resolution phenotype with that seen in patients with similar conditions/mutations. Throughout the project, we will share and disseminate our protocols, methods, and data to provide resources for use by the broader vision research community; this will be done using existing and newly-created online tools. A major strength of this application is the multidisciplinary team that has been assembled to take on this challenging project. The team brings the necessary complementary expertise required for model development, stem cell treatment, and evaluation of cell survival, integration, & function. This work will have a significant positive impact by providing not only validated disease models but also generalizable tools with which to create additional models in these and other species.

项目摘要/摘要。