The Development of Gene Therapy Using SIRT1 Signaling to Treat Chronic Glaucoma

项目摘要 300万美国人患有青光眼，其中只有一半的人知道自己患有青光眼。没有 治愈，10%的接受适当治疗的患者仍因眼内植入不足而出现视力丧失 压力控制和合规性。基因治疗作为一种干预措施，消除了患者主导的责任 解决这一全球负担的治疗方法。SIRT1信号通路在视觉保护中的作用 在视神经损伤过程中，使优秀的靶基因引入这一概念。本建议书要求 支持对开发细胞感兴趣的眼科医生获得职业发展指导奖- AAV靶向治疗青光眼的研究导师的专业知识涉及SIRT1在 基于其他视神经损伤模型的青光眼模型。在完成颁奖时 在此期间，候选人将获得提出、构建和执行实验所需的宝贵经验 从宾夕法尼亚大学目前的研究中获得的患者个性化数据的可视化研究。候选人的目标是 领导一个研究项目，通过使用基因疗法解决青光眼的治疗需求。 这套技术提供了一种新颖而合理的方法来治疗眼部其他细胞，以管理未来的青光眼。 该项目由宾夕法尼亚大学眼科的高级调查人员监督， 生理学，以及高级视网膜和眼科治疗中心(CAROT)将提供 应聘者职业发展的资源。该计划包括定期安排的会议，有条理的 课程工作、支持指导以及创新和可转换的科学目标。所有的导师都有 获得资金，以提供必要的资源，以确保提案中的目标取得成功。 本计划中概述的机构、导师和合作者致力于 早期的调查人员。这项计划将使候选人最终获得独立和专业知识 通过执行以下目标延长授权期： 目的1)明确视网膜节细胞SIRT1信号转导途径的分子机制。我们将确认SIRT1的作用 诱导多能干细胞(IPS)来源的视网膜神经节细胞在正常对照中的通路表达 细胞应激来自过氧化氢(H_2O_2)和拉伸室。 目的2)确定小分子激活剂和靶向AAV在SIRT1信号转导中的作用 在青光眼小鼠模型中的表达。我们将使用鼠标评估SIRT1激活的作用 慢性青光眼模型。 目的3)建立针对小胶质细胞和星形胶质细胞的细胞特异性腺病毒基因递送系统。我们 将开发将SIRT1靶基因输送到具有特定细胞的星形胶质细胞和小胶质细胞的技术 使用慢性青光眼活体模型的启动子。

PROJECT ABSTRACT Three million Americans have glaucoma with only half of them aware they carry the diagnosis. There is no cure and 10% of patients who with adequate treatment still have vision loss due to inadequate intraocular pressure control and compliance. Gene therapy as an intervention removes the responsibility of patient-driven treatments to address this global burden. The SIRT1 signaling pathway plays a role in visual preservation during optic nerve injury, making an excellent target gene to introduce this concept. This proposal requests support for a mentored career development award for an ophthalmologist with interest in developing cell- specific gene targeting using AAV to treat glaucoma. The expertise of the mentor involves the use of SIRT1 in a glaucoma model based on evidence from other models of optic nerve injury. At the completion of the award period the candidate will gain valuable experience required to propose, construct, and execute experiments in visual research from patient individualized data gained from current studies at UPenn. The candidate’s goal is to lead a research program that addresses a therapeutic need in glaucoma through the use of gene therapy. This skill set offers a novel and rational approach to treat other cells in eye to manage glaucoma in the future. This project is under the supervision of senior investigators at UPenn in the departments of Ophthalmology, Physiology, and the Center for Advanced Retinal and Ocular Therapeutics (CAROT) who will provide resources for candidate’s career development. The plan involves regular scheduled meetings, structured coursework, guidance for support, and innovative and translational scientific aims. All of the mentors have secured funding to provide necessary resources to ensure success of the aims in the proposal. The institution, mentors, and collaborators outlined in this proposal are dedicated to the research and growth of early investigators. This plan will position the candidate for independence and specialized expertise at the end of the award period through execution of the following AIMS: AIM 1) Define the molecular mechanism of SIRT1 pathway on RGCs. We will confirm the role of SIRT1 pathway expression in induced pluripotent stem cell (IPS) derived RGCs from unaffected control patients using cell stress from hydrogen peroxide (H2O2) and a stretch chamber. AIM 2) Determine the role of SIRT1 signaling with small molecular activators and targeted AAV expression in a mouse model of glaucoma. We will evaluate the role of SIRT1 activation using a mouse model of chronic glaucoma. AIM 3) Develop cell specific adenoviral specific gene delivery system for microglia and astrocytes. We will develop the technology to deliver SIRT1 target gene to astrocytes and microglial cells with specific cell promotors using in vivo models of chronic glaucoma.