Development of a gene therapy for inherited corneal disease using a lentivirus vector

使用慢病毒载体开发遗传性角膜疾病的基因疗法

• 批准号: 13470367
• 负责人: NISHIDA Koji
• 金额: $ 10.56万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470367/
• 关键词: gene therapy; corneal dystrophy; lentivirus vector; cell therapy; gone transfer; knockout mouse; CHST6; corneal stem cell; 培養角膜上皮シート; 斑状角膜ジストロフィ; トランスジェニックマウス; eGFP; 角膜発現特異的プロモーター; 角膜内皮

We investigated a gene therapy for inherited corneal diseases, so-called corneal dystrophies, using a lentivirus vector. Lentivirus vector can transduce non-dividing cells in vitro and in vivo, and mediate efficient transfer and sustained long-term expression of the transgene in varieties of tissues. In this project, we obtained the following results.(1) We have developed an enhancer-promotor region that drives a cornea-specific expression of a gene of interest.(2) We have created knockout mice of macular corneal dystrophy using a targeted destruction of CHST6 gene.(3) We have developed a transplantation approach of corneal epithelial sheets transduced with a lentivirus vector.(4) We have shown that lentivirus vector can transduce ex vivo corneal endothelial cells. The results suggest promising clinical capabilities for ex vivo gene therapy for the treatment for corneal dystrophies with a lentivirus vector.

我们研究了一种基因治疗遗传性角膜疾病，所谓的角膜营养不良，使用慢病毒载体。慢病毒载体在体外和体内均能转导非分裂细胞，介导转基因在多种组织中的高效转移和持续长期表达。在这个项目中，我们得到了以下结果：我们已经开发了一个增强子-启动子区域，它驱动感兴趣的基因的角膜特异性表达。(2)我们通过靶向破坏CHST6基因，建立了黄斑角膜营养不良的敲除小鼠。(3)我们开发了一种用慢病毒载体转导角膜上皮片的移植方法。(4)慢病毒载体可以转染离体角膜内皮细胞。研究结果表明，利用慢病毒载体进行离体基因治疗角膜营养不良具有良好的临床应用前景。

项目成果

Hayashida-Hibino S, Nishida K et al.: "The effect of TGF-betal on differential gene expression profiles in human corneal epithelium studied by cDNA expression array"Invest Ophthalmol Vis Sci. 42(8). 1691-1697 (2001)

Hayashida-Hibino S、Nishida K 等人：“TGF-β1 对通过 cDNA 表达阵列研究的人角膜上皮差异基因表达谱的影响”Invest Ophasemol Vis Sci。

西田 幸二: "遺伝子診断と分子機構(角膜疾患)"眼科. 44. 183-293 (2002)

Koji Nishida：“遗传诊断和分子机制（角膜疾病）”眼科。 44. 183-293 (2002)

Nakamura T, Nishida K et al.: "Changes in conjunctival clusterin expression in severe ocular surface disease"Invest Ophthalmol Vis Sci. 43. 1702-1707 (2002)

Nakamura T、Nishida K 等人：“严重眼表疾病中结膜簇蛋白表达的变化”Invest Ophasemol Vis Sci。

Dota A, Nishida K, Adachi W, Nakamura T, Koizumi N, Kawamoto S, Okubo K, Kinoshita S.: "An expression profile of active genes in human conjunctival epithelium."Exp Eye Res. 72. 235-241 (2001)

Dota A、Nishida K、Adachi W、Nakamura T、Koizumi N、Kawamoto S、Okubo K、Kinoshita S.：“人类结膜上皮中活性基因的表达谱。”Exp Eye Res。

Akama TO, Nakayama J, Nishida K, et al.: "Human corneal GlcNAc 6-O sulfotransferase and mouse intestinal GlcNAc 6-O sulfotransferase both produce keratan sulfate"J Biol Chem. 276. 16271-16278 (2001)

Akama TO、Nakayama J、Nishida K 等人：“人角膜 GlcNAc 6-O 磺基转移酶和小鼠肠道 GlcNAc 6-O 磺基转移酶均产生硫酸角质素”J Biol Chem。