Gene Delivery to the Trabecular Meshwork

基因递送至小梁网

• 批准号: 6644231
• 负责人: DOUGLAS J RHEE
• 金额: $ 14.78万
• 依托单位: WILLS EYE HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-02 至 2005-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6644231
• 关键词: 3T3 cells; Retroviridae; SDS polyacrylamide gel electrophoresis; alkaline phosphatase; biotechnology; cell cycle; clinical research; extracellular matrix; gene delivery system; gene therapy; genetic transcription; glaucoma; human tissue; immunocytochemistry; intraocular pressure; laboratory rabbit; lasers; metalloendopeptidases; migration inhibition factor; organ culture; polymerase chain reaction; reporter genes; trabecular meshwork; trabeculotomy; transfection /expression vector; western blottings

DESCRIPTION: (Applicant?s Abstract) The long-term goal of this research program is to deliver a gene product (i.e. gene therapy) which will increase extracellular matrix turnover specifically in the trabecular meshwork (TM) to lower intraocular pressure for the treatment of glaucoma. Two major obstacles to gene therapy in the trabecular meshwork are the lack of a tissue-specific delivery system and the lack of an effective gene which would lower IOP if successfully transfected. The general hypotheses are: (1) that a retroviral vector combined with laser-induced mitosis will deliver genetic material specifically to the trabecular meshwork and (2) that inacrophage migratory inhibitory (MIF) will cause an increase in extracellular matrix breakdown. These hypotheses will be tested by determining if the retroviral vector will transfer a reporter gene to dividing human and rabbit trabecular meshwork cells in culture. These studies will then be expanded to test the selectivity of the retroviral vector combined with laser-induced mitosis (RVCLIM) system in human and rabbit organ explant cultures and in rabbit eyes, in vivo. Additional studies will investigate MIF?s ability to upregulate the expression and activity of matrix metalloproteinases in cultured trabecular meshwork cells. If these experiments are successful, the RVLIM gene . delivery system could be used therapeutically and as a tool to study the biology of the TM. These studies would set the foundation for future evaluations of the function of MIF in the TM and MIF?s potential as a candidate for gene therapy for the treatment of glaucoma.

描述：（申请人的摘要）本研究计划的长期目标 是提供一种基因产品（即基因治疗），该产品将增加 细胞外基质周转特别是在小梁网（TM）中 降低眼压，用于治疗青光眼。两大障碍 小梁网中的基因治疗缺乏组织特异性 传递系统和缺乏有效基因会降低眼压，如果 成功转染。一般假设是：（1）逆转录病毒 载体与激光诱导有丝分裂相结合将传递遗传物质 特别是小梁网和（2）巨噬细胞迁移 抑制性（MIF）会导致细胞外基质分解增加。 这些假设将通过确定逆转录病毒载体是否会 将报告基因转移到分裂的人和兔小梁网细胞中 在文化中。这些研究随后将被扩展以测试 逆转录病毒载体联合激光诱导有丝分裂（RVCLIM）系统在人体内的应用 和兔器官外植体培养以及兔眼体内。额外的 研究将调查 MIF 上调表达和 培养的小梁网细胞中基质金属蛋白酶的活性。如果 这些实验是成功的，RVLIM基因。输送系统可以是 用于治疗并作为研究 TM 生物学的工具。这些 研究将为未来评估MIF的功能奠定基础 TM 和 MIF 作为基因疗法候选者的潜力 青光眼。