基因治疗降眼压的优化实验研究

Pharmaceutical treatment for lowering the intraocular pressure (IOP) is the major management of glaucoma. However, most of the existing medications exhibit problems including low efficacy, poor compliance and side effects. Preliminary studies in our group and literature showed that the extracellular enzyme C3 transferase enzyme (exoenzyme C3 transferase, C3) can effectively increase aqueous humor outflow facility of live animals or in organ cultured monkey eyes by disrupting actin cytoskeleton and its assocated cell-cell and cell-CEM interactins. Based on previous work, this project intends to construct a C3 gene eukaryotic expression system under the Ch3L1 trabecular meshwork tissue-specific promoter for transducing C3 into the rabbit eyes with polyethylenimine (PEI)-mediated gene transfer technology and in vivo ultrasound microbubble. A positive cross-experimental design will be used to explore the optimization of the experimental conditions of C3 gene delivery into the anterior chamber of rabbit eyes. The optimized experimental conditions are based on ideal IOP lowering effect and lightest side effects by evaluating results using intraocular pressure measurement and histological analysis. It will then provide optimized condition for C3 gene delivery into primate eyes. This study will provide experimental basis for the development of safe, effective and targeted gene therapy and bridge the gap between laboratory and clinical applications.

药物降眼压是治疗青光眼的主要手段。目前大多降眼压药均存在疗效不理想、长期用药依从性差和毒副作用等问题。课题组前期研究工作和文献均表明，外酶C3转移酶(exoenzyme C3 transferase, C3)以降解小梁细胞肌动蛋白细胞骨架、改变房水小梁通道构型从而降低房水排出阻力为作用机制，能有效提高动物或器官培养眼的房水流畅度，降低眼压。本课题在前期工作基础上深入进行C3基因降眼压的研究，拟构建含Ch3L1小梁细胞特异性启动子的C3基因真核表达载体，采用正交试验设计优化超声微泡联合聚乙烯亚胺介导的转基因技术将C3基因导入兔眼前房，通过眼压测量、组织病理分析等方法评价结果；在确定降眼压作用最好而副作用最轻的优化实验条件后，再在猴眼进行C3基因降眼压实验。本课题旨在研发安全有效的靶向性基因治疗降眼压方法，为青光眼基因治疗从最终实验室向临床应用过渡奠定基础。

药物降眼压是治疗青光眼的主要手段。目前大多降眼压药均存在疗效不理想、长期用药依从性差和毒副作用等问题。课题组前期研究工作和文献均表明，外酶 C3 转移酶 (exoenzyme C3 transferase, C3)以降解小梁细胞肌动蛋白细胞骨架、改变房水小梁通道构型从而降低房水排出阻力为作用机制，能有效提高动物或器官培养眼的房水流畅度，降低眼压。本课题在前期工作基础上深入进行 C3 基因降眼压的研究，拟构建含 Ch3L1 小梁细胞特异性启动子的 C3 基因真核表达载体，采用正交试验设计优化超声微泡联合聚乙烯亚胺介导 的转基因技术将 C3 基因导入兔眼前房，通过眼压测量、组织病理分析等方法评价结果；在确定降眼压作用最好而副作用最轻的优化实验条件后，再在猴眼进行 C3 基因降眼压实验。本课题旨在研发安全有效的靶向性基因治疗降眼压方法，为青光眼基因治疗从最终实验室向临床应用过渡奠定基础。