Myosin VIIa Gene Therapy

肌球蛋白VIIa基因治疗

• 批准号: 6710058
• 负责人: Xian-Jie Yang
• 金额: $ 13.5万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6710058
• 关键词: Lentivirus; Usher syndrome; biotechnology; fluorescence microscopy; gene delivery system; gene expression; gene mutation; gene therapy; genetic transcription; green fluorescent proteins; immunocytochemistry; laboratory mouse; myosins; nonhuman therapy evaluation; polymerase chain reaction; retinal pigment epithelium; transfection /expression vector; visual photoreceptor; western blottings; Lentivirus; Usher syndrome; biotechnology; fluorescence microscopy; gene delivery system; gene expression; gene mutation; gene therapy; genetic transcription; green fluorescent proteins; immunocytochemistry; laboratory mouse; myosins; nonhuman therapy evaluation; polymerase chain reaction; retinal pigment epithelium; transfection /expression vector; visual photoreceptor; western blottings

DESCRIPTION (provided by applicant): In Usher syndrome, deaf patients also develop retinitis pigmentosa. Usher syndrome type 1B has been associated with mutations in the myosin Vlla gene (MYO7A). In the mammalian eye, myosin VIla protein is located in the cilium of photoreceptor cells and in the apical region of the retinal pigmented epithelial (RPE) cells. Shaker1 mice also carry mutations in the myosin Vlla gene, and several mutant phenotypes have now been identified in the retinas of these mice. The proposed research aims to determine the efficacy of gene therapy for myosin Vlla deficiency in the shaker1 mouse. High titer lentiviruses co-expressing the human myosin Vlla protein and the green fluorescent protein (GFP) will be produced and injected into the sub-retinal space of newborn mice. The efficiency of viral infection and transgene transduction in photoreceptors and RPE cells will be determined by monitoring expression patterns of myosin Vlla and GFP. The rescuing effects of the viral mediated myosin Vlla expression will be determined by assaying previously identified mutant phenotypes in the retinas of shaker1 mice: e.g., the subcellular distribution of the pigment granules in the RPE cells, the distribution of rhodopsin in the photoreceptor connecting cilium, and the phagocytosis of the outer segment disks by the RPE cells. The proposed research will provide data directly relevant to the feasibility of gene therapy for the Usher 1 B syndrome in humans and to the application of lentivirus as a gene therapy vehicle for other inherited retinal diseases.

描述（由申请人提供）：在Usher综合征中，聋哑患者还会出现色素性视网膜炎。 1B型Usher综合征与肌球蛋白VLLA基因（MyO7A）中的突变有关。在哺乳动物的眼中，肌球蛋白VILA蛋白位于感光细胞的纤毛和视网膜色素上皮（RPE）细胞的顶端区域。 Shaker1小鼠还携带肌球蛋白VLLA基因中的突变，现在已经在这些小鼠的视网膜中鉴定出了几种突变表型。 拟议的研究旨在确定基因疗法对肌球蛋白VLLA缺乏症在Shaker1小鼠中的疗效。将产生并注入新生小鼠的视网膜下空间，并注入高滴度的慢滴病毒和绿色荧光蛋白（GFP）的共表达。光感受器和RPE细胞中病毒感染和转基因转导的效率将通过监测肌球蛋白VLLA和GFP的表达模式来确定。病毒介导的肌球蛋白VLLA表达的拯救作用将通过测定先前鉴定的Shaker1小鼠视网膜中的突变表型来确定：拟议的研究将提供与基因治疗对人类1 B综合征以及慢病毒作为其他遗传性视网膜疾病的基因治疗工具的可行性直接相关的数据。