Development of AAV-AIBP for neuroprotection in glaucoma

青光眼是世界范围内导致失明的主要原因。新出现的证据表明，神经胶质细胞炎症 是视网膜神经节细胞(RGC)死亡和视神经变性发病机制中的关键因素 青光眼的症状。Toll样受体-4(TLR4，炎症受体)基因多态性与三磷酸腺苷结合 盒转运体A1(ABCA1，细胞胆固醇转运体)与原发性开角型心脏病的风险有关 青光眼。我们的研究表明，TLR4在视网膜神经节细胞和胶质细胞中的表达增加，ABCA1的表达降低 人类青光眼视网膜。我们鉴定了分泌型apoA-I结合蛋白(AIBP；基因apoA1BP)是一个关键基因 细胞胆固醇代谢调节剂，通过依赖ABCA1的胆固醇控制TLR4的激活 TLR4的耗竭占据了炎症和活化细胞中的脂筏。与ABCA1类似，AIBP表达 在人类和小鼠青光眼中，胆固醇水平和炎症标志物水平降低，而炎症标志物水平上升 视网膜。Apoa1BP-/-小鼠视力受损，与野生型相比，视网膜TLR4增加 和IL-1β的表达，增强小胶质细胞的激活，并增加视网膜神经节细胞的死亡 眼压(IOP)。这些发现支持了AIBP在视网膜中恢复表达的理论基础 为青光眼患者提供持续的神经保护。在初步研究中，腺相关病毒 (AAV)-AIBP保护视网膜神经节细胞并改善青光眼实验小鼠模型的视觉功能障碍。在……里面 此外，重组AIBP蛋白促进线粒体功能，抑制炎症反应 在培养的Müller胶质细胞和小胶质细胞中，对升高的静水压力做出反应。基于我们之前的和 这些发现，我们建议发展基于AAV2-hAIBP的治疗，以减少视网膜神经炎症和 为接受标准眼压降低治疗的青光眼患者提供有效的神经保护。我们 提出以下里程碑驱动的具体目标：(1)优化AAV2-hAIBP并定义AIBP分布 (2)AAV2-hAIBP的体内效应和非GLP毒理学特性； 以及(3)进行启用IND的研究，并获得FDA批准进行第一次人体试验。该项目将结合 来自加州大学圣地亚哥分校、RAFT制药公司和NIH蓝图的专业知识和资源 神经治疗学网络-生物学。我们提出的研究将开发新的基因疗法来减少视网膜 该药可减少神经炎症和线粒体功能障碍，并为青光眼患者提供有效的神经保护。

Glaucoma is a leading cause of blindness worldwide. Emerging evidence suggests that glia-neuroinflammation is a critical element driving retinal ganglion cell (RGC) death and optic nerve degeneration in the pathogenesis of glaucoma. Gene polymorphisms in toll-like receptor-4 (TLR4, inflammatory receptor) and ATP-binding cassette transporter A1 (ABCA1, cellular cholesterol transporter) are linked to the risk of primary open-angle glaucoma. Our studies demonstrated increased TLR4 and reduced ABCA1 expression in RGCs and glia in human glaucomatous retina. We identified the secreted apoA-I binding protein (AIBP; gene APOA1BP) as a key regulator of cellular cholesterol metabolism, which controls TLR4 activation via ABCA1-dependent cholesterol depletion from TLR4 occupied lipid rafts in inflammatory and activated cells. Similar to ABCA1, AIBP expression was reduced, while cholesterol levels and inflammatory markers increased in human and mouse glaucomatous retinas. Apoa1bp-/- mice had compromised visual acuity and compared to wild type, had increased retinal TLR4 and IL-1β expression, augmented microglial activation, and increased RGC death in response to elevated intraocular pressure (IOP). These findings support the rationale for restoration of AIBP expression in the retina to provide sustained neuroprotection in patients with glaucoma. In preliminary studies, adeno-associated virus (AAV)-AIBP protected RGCs and ameliorated visual dysfunction in experimental mouse models of glaucoma. In addition, recombinant AIBP protein promoted mitochondrial function as well as inhibited inflammatory responses in cultured Müller glia and microglia in response to elevated hydrostatic pressure. Based on our previous and these findings, we propose development of AAV2-hAIBP based therapy to reduce retinal neuroinflammation and provide effective neuroprotection to glaucoma patients receiving standard-of-care IOP lowering treatment. We propose the following milestone-driven Specific Aims: (1) Optimize AAV2-hAIBP and define AIBP distribution and target engagement in retina; (2) Characterize the in vivo efficacy and non-GLP toxicology of AAV2-hAIBP; and (3) Conduct IND-enabling studies and obtain FDA approval for first-in-human trial. The project will combine expertise and resources from University of California, San Diego, RAFT Pharmaceuticals, and the NIH Blueprint Neurotherapeutics Network – Biologic. Our proposed studies will develop novel gene therapy to reduce retinal neuroinflammation and mitochondrial dysfunction and provide effective neuroprotection to glaucoma patients.