Novel model systems for the study of cone disorders and other heritable retinal diseases

用于研究视锥细胞疾病和其他遗传性视网膜疾病的新型模型系统

• 批准号: 10439118
• 负责人: JEFFREY A. ROGERS
• 金额: $ 15万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 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/10439118
• 关键词: Affect; Animal Model; Animals; Biological Models; Blindness; California; Cell Therapy; Cells; Characteristics; Communities; Cone; Cone dystrophy; DNA sequencing; Development; Disease; Electroretinography; FDA approved; Fluorescein Angiography; Fundus photography; Genes; Genetic; Goals; Heritability; Human; Inherited; Innovative Therapy; Leber' s amaurosis; Macaca; Macaca mulatta; Modeling; Morbidity - disease rate; Mutation; Optical Coherence Tomography; Patients; Phenotype; Primates; RPE65 protein; Research; Retina; Retinal Degeneration; Retinal Diseases; Retinitis Pigmentosa; Rod; Series; Speed; Thinness; Translations; Vision research; Visual impairment; Work; base; behavior observation; disorder of macula of retina; effective therapy; efficacy testing; gene function; gene replacement therapy; gene therapy; genetic testing; human disease; human model; macula; nonhuman primate; novel; novel therapeutics; population survey; protein function; response

Project Summary/Abstract Retinal degeneration diseases are a common cause of untreatable blindness worldwide, affecting the lives of millions. The only FDA-approved treatment for these disorders is gene therapy for specific RPE65 mutations that cause Leber’s congenital amaurosis and retinitis pigmentosa. One major limitation to the development of effective therapies is the use of animal models that poorly replicate the human condition. Particularly for cone disorders, studies that use animals with a rod-dominant retina and no true macula have substantive limitations. By contrast, the cone-rich macula of nonhuman primates (NHP) closely mirrors that of the human retina. Consequently, well-defined NHP models of heritable retinal diseases, particularly cone disorders, that are more predictive of human conditions are necessary to more efficiently advance new therapies. We propose to develop a series of novel and spontaneous NHP models of human inherited retinal diseases. Behavioral observations of rhesus macaques (Macaca mulatta) at the California National Primate Research Center identified a series of macaques that displayed apparent visual impairment. Genetic testing showed that four animals are homozygous for a damaging mutation in the PDE6C gene, which has previously been associated with cone dystrophy in humans. Scotopic and photopic full-field electroretinograms performed on macaques homozygous for the PDE6C mutation demonstrated a relatively normal rod response but no cone response whatsoever. A subtle but characteristic bullseye maculopathy was identified using fundus photography, blue autofluorescence, and fluorescein angiography with concurrent foveal thinning using spectral-domain optical coherence tomography. Our genetic survey of this population also identified macaques with mutations in 7 other human retinal disease genes that are predicted to severely damage gene or protein function, pointing to possible additional new models. To develop the PDE6C primate model of cone dystrophy, and make this and other new NHP models available to the vision research community, we propose four Specific Aims: 1) to identify and genetically characterize new NHP models of human retinal disease via DNA sequencing, 2) to perform complete ophthalmic phenotyping of NHP models of retinal disease, 3) to breed a colony of NHPs with PDE6C cone dystrophy and 4) to compare cell-based and gene replacement therapies in macaques with PDE6C cone dystrophy mutations. Successful completion of this work will produce a well-characterized new animal model of inherited cone dystrophy with significantly greater similarity to human disease than existing models, thus providing substantially better translation to subsequent human trials. In addition, affected animals will be made available to the wider vision research community, and other new models with similar potential will be identified.

项目概要/摘要 视网膜变性疾病是全世界无法治愈的失明的常见原因，影响着人类 数百万人的生命。 FDA 批准的治疗这些疾病的唯一方法是针对特定疾病的基因治疗 RPE65 突变导致莱伯先天性黑蒙和色素性视网膜炎。一个专业 开发有效疗法的限制是使用复制效果不佳的动物模型 人类状况。特别是对于视锥细胞疾病，使用具有视杆细胞优势视网膜的动物进行的研究 并且没有真正的黄斑具有实质性的局限性。相比之下，非人类的富含视锥细胞的黄斑 灵长类动物（NHP）与人类视网膜非常相似。因此，明确定义的 NHP 模型 遗传性视网膜疾病，特别是视锥细胞疾病，更能预测人类的状况 更有效地推进新疗法是必要的。我们建议开发一系列新颖且 人类遗传性视网膜疾病的自发 NHP 模型。恒河猴的行为观察 加州国家灵长类动物研究中心的猕猴（Macaca mulatta）发现了一系列 表现出明显视力障碍的猕猴。基因测试表明，四只动物 PDE6C 基因中存在破坏性突变的纯合子，该突变此前曾被认为与 人类锥体营养不良。对猕猴进行暗视和明视全视野视网膜电图 PDE6C 突变纯合子表现出相对正常的视杆细胞反应，但没有视锥细胞反应 无论如何回应。使用眼底检查发现了一种微妙但特征性的牛眼黄斑病变 摄影、蓝色自发荧光和荧光素血管造影，同时使用中央凹变薄 谱域光学相干断层扫描。我们对该人群的基因调查还发现 猕猴体内 7 种其他人类视网膜疾病基因发生突变，预计会严重损害视网膜 基因或蛋白质功能，指出可能的其他新模型。开发 PDE6C 灵长类动物 视锥细胞营养不良模型，并使该模型和其他新的 NHP 模型可用于视觉研究 社区，我们提出了四个具体目标：1）识别新的 NHP 模型并对其进行遗传表征 通过 DNA 测序分析人类视网膜疾病，2) 对 NHP 进行完整的眼科表型分析 视网膜疾病模型，3) 培育具有 PDE6C 视锥细胞营养不良的 NHP 群体，4) 进行比较 针对 PDE6C 视锥细胞营养不良突变的猕猴进行基于细胞和基因替代疗法。 这项工作的成功完成将产生一个特征良好的遗传性新动物模型 与现有模型相比，锥体营养不良与人类疾病的相似性明显更高，因此 为随后的人体试验提供更好的转化。此外，受影响的动物将 可供更广泛的视觉研究界以及具有类似潜力的其他新模型使用 将被识别。