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: 10631293
• 负责人: Joseph Carroll
• 金额: $ 8.62万
• 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 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10631293
• 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。 NEI 的 Audacious Goal Initiative（2012 年启动）提出了“恢复可用视力”的挑战 “通过再生眼睛和视觉系统中的神经元和神经连接。” 对新疗法的亲和力、当前的资源和技术差距阻碍了许多疗法的转化 其中一个差距与共享人类视网膜关键特征的动物模型的可用性有关。 解剖学以及忠实模拟患者所见机制和过程的疾病模型 视网膜变性（可能适合再生疗法的致盲疾病）。 容易获得的视锥细胞优势哺乳动物模型是阻碍努力的主要技术差距 开发和评估视网膜再生治疗策略我们建议推进两项。 有前途的模型系统比更广泛使用的模型系统更接近人类视觉解剖结构和功能 第一个是 13 线地松鼠 (13-LGS)：一种昼夜性锥体啮齿动物（约 85%）。 第二种是树鼩：一种非视觉信息的大脑区域。 啮齿动物、类似灵长类的哺乳动物，也是视锥细胞占主导地位的（~95% 视锥细胞）这些模型已用于研究。 视觉转导 (13-LGS)、外节形态发生、脱落和冬眠期间的重塑 (13- LGS）、视锥双极细胞电路（13-LGS）、近视（树鼩）和中央视觉处理（树鼩）。 然而，它们用作翻译支持模型来评估再生的生存和整合 视锥细胞的光感受器受到限制，主要是由于缺乏对这些视锥细胞进行基因操作的工具。 我们建议将这些物种视为与疾病相关的物种。 通过以下具体目标建立模型：（1）开发、优化和验证成像方法和功能 13-LGS 和树鼩的测定；(2) 从 iPSC 生成 13-LGS 和树鼩锥光感受器 体外；（3）为 13-LGS 和树鼩建立 rAAV 介导的视网膜变性模型；（4）启用； 人类疾病种系转基因13-LGS模型；（5）测试和优化移植13-LGS的整合 LGS、树鼩和正常和退化的 13-LGS 和树鼩视网膜 A 中源自人类 iPSC 的视锥细胞。 该提案的主要特点是通过比较这些模型的细胞分辨率表型来验证它们 在整个项目中，我们将分享和传播在具有类似病症/突变的患者中看到的情况。 我们的协议、方法和数据为更广泛的视觉研究社区提供资源； 将使用现有的和新创建的在线工具来完成该应用程序的主要优势是 已组建多学科团队来承担这一具有挑战性的项目。 模型开发、干细胞治疗和细胞评估所需的必要补充专业知识 这项工作不仅提供经过验证的结果，还将产生重大的积极影响。 疾病模型，以及在这些物种和其他物种中创建其他模型的通用工具。