Advancing novel therapies for optic neuropathy with a nonhuman primate model

项目总结 视神经疾病是世界范围内常见的致盲原因，影响数百万人的生活，而且缺乏有效的治疗方法 恢复视力的治疗方法。常染色体显性遗传性视神经萎缩是一种遗传性视神经病变， 全世界每10万人中就有3人受到影响，导致视力障碍，而且没有治疗方法。它主要是由于 视神经萎缩1(OPA1)基因突变，该基因编码一种关键的线粒体动力蛋白相关蛋白 线粒体的稳定性和能量的产生。开发有效的治疗方法的一个主要限制因素 视神经疾病是利用动物模型来复制人类的情况。尤其是对于光学 神经和RGC障碍，研究将受益于使用非人类灵长类动物(NHP)与视神经和 视网膜解剖学、生理学和病理学，与人类的视网膜解剖、生理和病理学密切相关。因此，定义明确的 更能预测人类状况的NHP视神经病变模型对于有效地 先进的新疗法。我们鉴定了猕猴的杂合子和纯合子的错义突变 与OPA1相比，OPA1表现为视神经头苍白和视网膜神经纤维层变薄 野生型对照；这些发现与人类患者的ADOA一致。我们将充分定义这种NHP模式 并确定其在5年内对区域GC结构和功能的纵向影响。具体来说， 我们将使用视网膜电图、视网膜黄素蛋白来评估视网膜功能障碍的发生和发展。 荧光法和目测法。重要的是，我们将把临床发现与详细的转录本相关联， 组织学和免疫组织化学数据，以全面描述这种NHP模型的ADOA。 来自NHP的ADOA转录组很有可能识别与RGC有关的新基因和途径 病理学和神经退行性变以及验证先前涉及的途径，从而揭示了新的 治疗靶点。我们还将进行详细的组织学、免疫组织化学和超微结构分析。 为了评估RGC胞体、轴突和树突以及它们在黄斑中的线粒体大小和数量， 黄斑乳头状束和视网膜的周边。最后，通过选择性繁殖受ADOA影响的NHP， 我们将为未来的研究产生OPA1突变的杂合子和纯合子猕猴。 为了使这种新的NHP视神经病变模型可用于治疗测试，我们提出了三个具体目标： 1)确定ADOA的NHP模型的形态特征和表型谱，2)确定 OPA1基因A8S突变对非霍奇金淋巴瘤RGC功能的影响；3)检测mRNA和蛋白表达 在受ADOA影响的NHP的RGCs中。曾经最具预测性的疾病终点和足够多的动物 在ADOA被确认后，我们将继续进行更多的病因研究和新的治疗策略。这 ADOA的综合NHP模型将是视觉科学和 神经退行性疾病社区。

PROJECT SUMMARY Optic neuropathies are common causes of blindness worldwide, affect the lives of millions, and lack effective treatments to restore vision. Autosomal dominant optic atrophy (ADOA) is an inherited optic neuropathy that affects ~3 per 100,000 people worldwide, results in vision impairment, and has no treatment. It is primarily due to mutations in the optic atrophy 1 (OPA1) gene, which encodes a mitochondrial dynamin-related protein critical for mitochondrial stability and energy production. A major limitation to the development of effective therapies for optic neuropathies is the use of animal models that poorly replicate the human condition. Particularly for optic nerve and RGC disorders, studies would benefit from the use nonhuman primates (NHP) with optic nerve and retinal anatomy, physiology and pathology, which closely mirrors that of humans. Consequently, well-defined NHP models of optic neuropathy that are more predictive of human conditions are necessary to efficiently advance new therapies. We identified rhesus macaques heterozygous and homozygous for a missense mutation in OPA1 that demonstrate optic nerve head pallor and thinning of the retinal nerve fiber layer in comparison to wildtype controls; these findings are consistent with ADOA in human patients. We will fully define this NHP model of ADOA and determine its impact on RGC structure and function longitudinally over a 5-year period. Specifically, we will assess the onset and progression of retinal dysfunction utilizing electroretinography, retinal flavoprotein fluorescence and visual testing. Importantly, we will correlate the clinical findings with detailed transcriptomic, histologic, and immunohistochemical data for a comprehensive characterization of this NHP model of ADOA. The ADOA transcriptome from NHPs is highly likely to identify novel genes and pathways involved in RGC pathology and neurodegeneration as well as validate previously implicated pathways thus revealing new therapeutic targets. We will also perform detailed histological, immunohistochemical and ultrastructural analyses to assess RGC soma, axons and dendrites as well as their mitochondrial size and number in the macula, papillomacular bundle, and periphery of the retina. Finally, through selective breeding of ADOA-affected NHPs, we will generate a supply of macaques heterozygous and homozygous for the OPA1 mutation for future study. To make this new NHP optic neuropathy model available for therapeutic testing, we propose three Specific Aims: 1) To define the morphologic features and phenotypic spectrum of a NHP model of ADOA, 2) to determine the impact of the OPA1 A8S mutation on RGC function in NHPs, and 3) to determine mRNA and protein expression in RGCs of ADOA-affected NHPs. Once the most predictive endpoints of disease and sufficient animals with ADOA are identified, we will pursue additional studies of etiopathogenesis and novel therapeutic strategies. This comprehensive NHP model of ADOA will be a highly valuable resource for the vision science and neurodegenerative disease communities.