The role of impaired mitophagy and mitochondrial dysfunction in glaucomatous neurodegeneration

项目摘要/摘要： 原发性开角型青光眼(POAG)是青光眼最常见的一种形式，其特点是进行性 视网膜神经节细胞(RGC)及其轴突丢失，导致不可逆转的视力丧失。高位眼内 压力(IOP)是POAG的主要危险因素。不幸的是，潜在的病理机制 眼压引起的青光眼神经变性的原因仍不清楚。这样做的长期目标是 建议描绘调控眼压诱导的青光眼神经变性的分子途径和 开发一种有效的青光眼治疗策略。为此，我们开发了一种新型的糖皮质激素 (GC)诱导和霉菌素相关的POAG小鼠模型，复制人类POAG表型。 重要的是，我们发现了线粒体吞噬功能受损、线粒体受损和炎症。 人和小鼠青光眼视神经的免疫细胞。根据我们的初步数据，我们 建议：1)观察眼压对线粒体吞噬功能损害和聚集的影响 利用小鼠和人的POAG，2)检测吞丝分裂功能受损在青光眼神经变性中的作用 以及，3)进一步确定增强有丝分裂是否减轻了神经元的变性和防止了RGC的丢失 POAG的小鼠模型。在指导阶段，我将建立小鼠模型来研究POAG中的有丝分裂现象 包括丝裂原报告基因转基因mt-keima小鼠、RGC特异性Parkin和ATG5条件性基因敲除 在Gulab Zode博士(小梁慢性内质网应激和自噬方面的专家)的指导下的小鼠 网络)。在丹尼斯·英曼博士(线粒体新陈代谢领域的专家)的合作下，我将 加深对POAG小鼠模型线粒体功能障碍的认识。指导阶段将 还将辅以Abbot Clark博士(著名的青光眼专家)的培训，他将提供 对人体组织的帮助，以及为我的独立职业生涯提供的额外培训。此外，定期 与在协助年轻调查人员方面拥有丰富经验的保拉·格雷戈里博士的会面将对我有所帮助 发展独立事业。在独立阶段，我们将研究吞噬有丝分裂在 炎性神经变性和确定是否诱导有丝分裂(尿素素A/放线菌素/二甲双胍/ Parkin过度表达)挽救了GC或myocin相关的POAG。此外，在指导阶段，我 将定期与我的顾问委员会举行会议，参加科学会议，并继续我的 职业发展。我处于理想的环境中，适合进行拟议的研究和我的职业发展 佐德博士在北德克萨斯眼科研究所拥有一流的设施， 与著名科学家瓦尔·谢菲尔德博士、约翰·胡尔曼博士和凯文·帕克博士合作。这会有帮助的 我要建立良好的合作关系，学习新技术，在一口井上建立独立的研究实验室- 建立了学术机构。

PROJECT SUMMARY/ABSTRACT: Primary open angle glaucoma (POAG), the most common form of glaucoma, is characterized by progressive loss of retinal ganglion cells (RGCs) and their axons, leading to irreversible vision loss. Elevated intraocular pressure (IOP) is the major risk factor for POAG. Unfortunately, the underlying pathological mechanisms responsible for IOP-induced glaucomatous neurodegeneration still remain unclear. The long-term goal of this proposal is to delineate the molecular pathways governing IOP-induced glaucomatous neurodegeneration and to develop an effective glaucoma treatment strategy. To this end, we have developed a novel glucocorticoid (GC)-induced and myocilin-associated mouse models of POAG, replicating human POAG phenotypes. Importantly, we have identified impaired mitophagy, accumulation of damaged mitochondria and inflammatory immune cells in the optic nerve of both human and mouse glaucoma. Based upon our preliminary data, we propose to: 1) examine the effect of IOP on mitophagy impairment and accumulation of damaged mitochondria using mouse and human POAG, 2) examine the role of impaired mitophagy on glaucomatous neurodegeneration and, 3) further identify whether enhancing mitophagy alleviates neurodegeneration and prevent RGC loss in mouse models of POAG. In the mentored phase, I will establish mouse models to study mitophagy in POAG including mitophagy reporter transgenic Mt-Keima mice, RGC-specific Parkin and ATG5 conditional knockout mice under the guidance of Dr. Gulab Zode (an expert in chronic ER stress and autophagy in trabecular meshwork). In collaboration with Dr. Denise Inman (an expert in the field of mitochondrial metabolism), I will enhance my understanding of mitochondrial dysfunction in mouse models of POAG. The mentored phase will also be supplemented by training with Dr. Abbot Clark (well-known leading glaucoma expert), who will provide assistance with human tissues as well as an additional training for my independent career. Furthermore, regular meetings with Dr. Paula Gregory who has tremendous experience in assisting young investigators will help me to develop independent career. During the independent phase, we will examine role of mitophagy on inflammatory neurodegeneration and determine whether inducing mitophagy (Urolithin A/Actinonin/Metformin/ Parkin overexpression) rescues GC or myocilin-associated POAG. Additionally, during the mentored phase, I will be having regular meetings with my advisory committee, attend scientific conferences, and continue my career development. I am in the ideal environment for the proposed research and for my career development as Dr. Zode has an established state-of-the-art facilities at the North Texas Eye Research Institute, and collaborations with renowned scientists, Dr. Val Sheffield, Dr. John Hulleman and Dr. Kevin Park. This will help me to set-up good collaborations, learn new techniques, and build an independent research laboratory at a well- established academic institution.