mTOR-mediated Signaling Pathway in Aging of the Retinal Pigment Epithelium

项目摘要 申请人研究计划的长期目标是探索分子机制 年龄相关性黄斑变性（AMD）的病因，并开发新的靶向治疗药物 战略布局此K99/R 00补助金将促进申请人成为独立的 范德比尔特眼科研究所的研究员，该研究所拥有丰富的科学合作环境， 培养年轻科学家的职业发展。AMD是美国老年人失明的主要原因。 大多数AMD患者具有萎缩（干性）形式的疾病，并且仅具有有限的治疗选择， 现在的时间萎缩性AMD可能是由于基因/环境相互作用导致进行性AMD， 视网膜色素上皮（RPE）变性。老龄化是最明确的环境风险因素 关于AMD我们假设哺乳动物雷帕霉素靶蛋白（mTOR）介导的信号通路 在控制RPE的老化过程中起关键作用。这一假设得到了最近 文献数据表明，mTOR在整合各种环境信号和连接 它们改变了组织功能和生物体的寿命。在我们的初步研究中，我们发现雷帕霉素 逆转体外原代人RPE细胞的衰老表型。为了进一步验证我们的假设，我们 在本申请中提出了三个具体目标。目的1是确定衰老过程如何调节 RPE中的mTOR通路。目的2是确定调节mTOR信号传导如何影响视网膜色素上皮细胞的老化。 通过调节自噬的潜在机制在体外进行。目的3是确定mTOR是否调节衰老 使用在视网膜中发展AMD样表型的SOD 1敲除小鼠在体内观察RPE。期间 在指导阶段，申请人将在指导人员的监督下进行拟议的实验。 该委员会由具有AMD病因学和发病机制方面的专业知识的导师组成（Paul博士 斯滕贝格）、视网膜神经变性的动物模型（大卫·卡尔金斯博士）和mTOR/自噬（大卫·卡尔金斯博士）。 陆波）。委员会成员将定期举行会议，监察研究进展，并协助申请人 让她的事业走向独立R 00阶段的研究将测试潜在的治疗方法， mTOR抑制剂在与AMD相关的动物模型中的作用。这些研究的结果将提供新的 关于RPE和视网膜的老化和年龄相关性变性的机制信息。

PROJECT SUMMARY The long term goal of the Applicant's research program is to explore the molecular mechanisms underlying the etiology of age-related macular degeneration (AMD) and develop novel target-based therapeutic strategies. This K99/R00 grant will facilitate the transition for the Applicant to become an independent investigator at the Vanderbilt Eye Institute, which has a rich environment of scientific collaboration and nurturing career development of junior scientists. AMD is the leading cause of blindness in elderly Americans. The majority of AMD patients has atrophic (dry) form of the disease and has only limited treatment options at the present time. Atrophic AMD is likely resulted from gene/environmental interaction causing progressive degeneration of the retinal pigment epithelium (RPE). Aging is the most well defined environmental risk factor of AMD. We hypothesize that the mammalian target of rapamycin (mTOR)-mediated signaling pathway plays key roles in controlling the aging process of the RPE. The hypothesis is supported by recent literature data suggesting that mTOR plays key roles in integrating various environmental signals and linking them to altered tissue function and organism's life span. In our preliminary studies, we found that rapamycin reversed the senescent phenotype of primary human RPE cells in vitro. To further test our hypothesis, we have proposed three specific aims in this application. Aim 1 is to determine how the aging process regulates mTOR pathways in the RPE. Aim 2 is to determine how modulating the mTOR signaling affects RPE aging in vitro by a potential mechanism of regulating autophagy. Aim 3 is to determine whether mTOR regulates aging of the RPE in vivo using SOD1 knockout mice which develop AMD-like phenotype in the retina. During the mentored phase, the Applicant will conduct the proposed experiments under the supervision of a mentoring committee, which is consisted of mentors with expertise in AMD etiology and pathogenesis (Dr. Paul Sternberg), animal models of neurodegeneration in the retina (Dr. David Calkins) and mTOR/autophagy (Dr. Lu Bo). Members of the committee will meet regularly, monitor the research progress and assist the Applicant to advance her career towards independence. Research at the R00 phase will test the potential therapeutic effects of mTOR inhibitors in an animal model relevant to AMD. Results from these studies will provide novel mechanistic information on aging and age-related degeneration of the RPE and retina.