Prevention of Retinal Degeneration by Crystallins in Experimental Uveitis

描述（由主要研究者提供）：眼内炎症，通常称为葡萄膜炎，是视网膜感光细胞凋亡导致失明的主要原因。非常类似于人葡萄膜炎的葡萄膜炎的稳健动物模型是实验性自身免疫性葡萄膜视网膜炎（EAU）。其特征在于两个阶段：早期，免疫后5 - 7天（p.i.）与葡萄膜原抗原，晚期/扩增阶段，12 - 14天p.i.在晚期阶段，巨噬细胞/中性粒细胞浸润视网膜，并且在组织学上注意到光感受器损伤。在白细胞浸润之前的早期阶段，揭示了视网膜中的Th1和Th17驱动的细胞因子、来自线粒体氧化应激的光感受器中的细胞色素c（cyto c）的硝化以及硝化的cyto c释放到胞质溶胶中。已知这种细胞因子释放会引起细胞凋亡。然而，凋亡不是早期EAU的特征，这表明EAU视网膜中存在防止感光细胞凋亡的保护机制。目前的初步研究表明，这种保护是由1A晶体蛋白上调提供的，这是光感受器所特有的，不受公认的抗凋亡蛋白（包括1B晶体蛋白、热休克蛋白27和其他热休克蛋白）介导。值得注意的是，具有早期EAU的1A晶体蛋白敲除小鼠在视网膜中显示出显著的细胞凋亡;这种细胞凋亡在具有EAU的野生型小鼠中完全不存在。这些数据揭示了1A晶体蛋白在防止视网膜细胞凋亡中的意义。此外，这表明与其他组织不同，光感受器选择性地利用1A晶体蛋白来预防早期EAU中的氧化应激介导的损伤，这是在光感受器中观察到的一种新的保护机制。基于我们的初步研究，我们假设1）1A晶体蛋白的上调是光感受器响应线粒体氧化应激的特征; 2）上调的1A晶体蛋白与氧化应激诱导的硝化细胞色素c相互作用，阻断半胱天冬酶的激活，并防止光感受器的凋亡。这些假设将通过三个特定目的进行检验：1）确定EAU期间视网膜中1A晶体蛋白上调的免疫机制; 2）确定EAU中1A晶体蛋白对感光细胞凋亡的预防; 3）确定EAU中iNOS对感光细胞氧化应激的诱导。对EAU中1A晶体蛋白上调的免疫机制及其保护性抗凋亡功能的深入分析将通过阐明晶体蛋白上调不仅在葡萄膜炎中而且在其他氧化应激介导的视网膜变性中在光感受器保护中的重要性来增强我们的理解。此外，目前的建议表明，1A晶体蛋白可以是一个有前途的治疗剂，用于预防氧化应激介导的感光细胞损伤。这种氧化损伤与各种致盲疾病有关，包括年龄相关性黄斑变性、视网膜静脉阻塞和青光眼。公共卫生相关性α A晶体蛋白是存在于眼睛和其他组织中的独特蛋白质。缺乏这种蛋白质会导致失明，补充这种蛋白质可以预防几种致盲疾病。

DESCRIPTION (provided by principal investigator): Intraocular inflammation, commonly referred to as uveitis, is a leading cause of blindness from retinal photoreceptor cell apoptosis. A robust animal model of uveitis that closely resembles human uveitis is experimental autoimmune uveoretinitis (EAU). It is characterized by two phases: the early phase, 5-7 days postimmunization (p.i.) with an uveitogenic antigen, and the late/amplification phase, 12-14 days p.i. During the late phase, macrophages/neutrophils infiltrate the retina, and photoreceptor damage is noted histologically. The early phase, preceding the leukocyte infiltration, reveals Th1- and Th17-driven cytokines in the retina, nitration of cytochrome c (cyto c) in the photoreceptors from mitochondrial oxidative stress, and the release of nitrated cyto c into the cytosol. Such cyto c release is known to cause apoptosis. However, apoptosis is not a feature of early EAU, suggesting the presence of a protective mechanism(s) in the EAU retina that prevents photoreceptor apoptosis. Current preliminary studies suggest that the protection is offered by 1A crystallin upregulation and that this is unique to the photoreceptors and are not mediated by well-recognized anti-apoptotic proteins, including 1B crystallin, heat shock protein 27 and other heat shock proteins. Significantly, 1A crystallin knockout mice with early EAU show prominent apoptosis in the retina; such apoptosis is totally absent in the wild type mice with EAU. The data reveal the significance of 1A crystallin in preventing apoptosis in the retina. Moreover, it indicates that unlike other tissues, the photoreceptors utilize 1A crystallins selectively in the prevention of oxidative stress-mediated damage in early EAU, a novel protective mechanism observed in the photoreceptors. Based on our preliminary studies we hypothesize that 1) upregulation of 1A crystallin is a feature of the photoreceptors in response to mitochondrial oxidative stress; 2) upregulated 1A crystallin interacts with the oxidative stress-induced nitrated cyto c, blocking the activation of caspases, and preventing apoptosis of photoreceptors. These hypotheses will be tested by three specific aims: 1) Determine the immune mechanism of 1A crystallin upregulation in the retina during EAU; 2) Determine prevention of photoreceptor apoptosis by 1A crystallin in EAU and 3) Determine photoreceptor oxidative stress induction by iNOS in EAU. In-depth analysis of the immune mechanism(s) of 1A crystallin upregulation in EAU and its protective anti- apoptotic function will enhance our understanding, by clarifying the importance of crystallin upregulation in photoreceptor protection not only in uveitis, but in other oxidative stress-mediated retinal degenerations as well. Moreover the current proposal suggests that 1A crystallin can be a promising therapeutic agent for preventing oxidative stress-mediated photoreceptor damage. Such oxidative damage has been implicated in various blinding diseases, including age-related macular degeneration, retinal vein occlusion, and glaucoma. PUBLIC HEALTH RELEVANCE AlphaA crystallin is a unique protein present in the eye and other tissues. Lack of this protein can cause blindness, and supplement of this protein can prevent several blinding diseases.