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，其中只有一半人知道自己患有青光眼。没有 10%的患者在充分治疗后仍因眼内不充分而导致视力丧失。 压力控制和顺应性。基因治疗作为一种干预措施， 治疗，以解决这一全球负担。SIRT1信号通路在视觉保护中发挥作用 在视神经损伤过程中，使一个很好的目标基因引入这一概念。该提案要求 为一名有兴趣开发细胞的眼科医生提供辅导职业发展奖， 使用AAV的特异性基因靶向治疗青光眼。导师的专业知识涉及SIRT 1的使用， 一种基于其他视神经损伤模型证据的青光眼模型。颁奖结束时 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视觉研究。候选人的目标是 领导一项研究计划，通过使用基因疗法来解决青光眼的治疗需求。 这一技能组合提供了一种新颖而合理的方法来治疗眼睛中的其他细胞，以在未来控制青光眼。 该项目是在宾夕法尼亚大学眼科部门的高级研究人员的监督下， 高级视网膜和眼科治疗中心（CAROT）将提供 候选人职业发展的资源。该计划包括定期安排会议，结构化 课程，指导支持，创新和转化的科学目标。所有的导师 获得拨款，以提供所需资源，确保成功实现建议书中的目标。 本提案中概述的机构，导师和合作者致力于研究和发展 早期的研究者该计划将定位候选人的独立性和专业知识在年底 通过执行以下AIMS， 目的1）阐明SIRT 1通路对RGCs的作用机制。我们将确认SIRT1的作用 在来自未受影响的对照患者的诱导的多能干细胞（IPS）衍生的RGC中，使用 来自过氧化氢（H2O2）的细胞应激和拉伸室。 目的2）用小分子活化剂和靶向AAV确定SIRT1信号传导的作用 在小鼠青光眼模型中的表达。我们将使用小鼠评估SIRT1激活的作用。 慢性青光眼模型。 目的3）建立小胶质细胞和星形胶质细胞特异性腺病毒基因载体系统。我们 将开发将SIRT1靶基因递送到具有特定细胞的星形胶质细胞和小胶质细胞的技术， 使用慢性青光眼的体内模型。

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.