New Methods of Gene Transfer to the Retina

基因转移到视网膜的新方法

• 批准号: 12557145
• 负责人: NAKAZAWA Mitsuru
• 金额: $ 8万
• 依托单位: Hirosaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557145/
• 关键词: retinitis pigmentosa; gene transfer; gene therapy; electroporation; conjunctiva; glaucoma; retina

Retinitis pigmentosa is a complex of hereditary progressive retinal degenerations that is nominated as the third commonest cause of legal blindness in adult population in Japan with an incidence of 1 out of 3,500 people, and therefore is an important disease in terms of the policy against blindness. Gene therapy, retinal transplantation and visual prosthesis have been currently studied by many researchers in the world, and expected to be developed as effective treatment. In this study, we investigated the technical possibility and effect of in vivo gene transfer to the retina by electroporation as a method of gene therapy for retinal degeneration. To test the technical possibility, we employed pars plana vitrectomy on the rabbit eyes to approach the retina, injected DNA solution in the subretinal space, and applied electroporation with a needle electrodes. The result indicated that this method could transfer DNA only in a small area of the retina, which might be insufficient to cause t … More herapeutic effects. It also indicated that we needed to develop another type of electrodes such as cup-shaped one. As the second part of the study, we studied the expression and effect of electroporatic gene transfer to the ocular tissue. First of all, we employed conjunctival tissue as a target of gene transfer prier to the retina, because it seemed easier to transfer DNA to the conjunctiva than to the retina. The result showed that the green fluorescent protein transferred to the conjunctiva by electroporation had been apparently expressed in the target tissue 30 days after transfer. Then we tried to transfer metalloproteinase 3 cDNA to the conjunctival flap during trabeculectomy on the rabbit eyes to examine the effect of the gene on intraocular pressure in the postoperative period. The result indicated that postoperative intraocular pressure of the eye treated with gene transfer showed as low as the eye treated by trabeculectomy with MMC, and that there was no pathological reaction in the area where DNA had been injected. All these results have suggested that we successfully transferred DNA fragments to the conjunctiva prier to the retinal, that it is possible for us to perform gene transfer to the retina using electroporation with some modification, and that gene therapy can be applied to glaucoma filtration surgery. Less

色素性视网膜炎是遗传性渐进性视网膜变性的综合，这是日本ADU LT人口中法律盲目的第三名，其中有3500人中有1人，这是对盲人政策的进口疾病在世界上，许多研究人员都在研究治疗，视网膜超违然和视觉假体，并在这项研究中有效治疗。为了进行视网膜变性，我们在兔子眼上使用PARS Plana玻璃体切除术，将视网膜下的DNA溶液注射，并用针射线涂抹电极。不足会导致t ...更多的效果。因为它似乎将DNA转移到结膜上，因此结果表明，在转移后30天，在靶Ssue中表达了绿色荧光，然后在兔子眼中转移了金属蛋白酶3 cDNA，然后将其转移到兔子眼上。为了检查术中的术中，基因转移的作用是通过小梁切除术与MMC进行的，并且在该地区没有病理真正的真实现实主义。使用电穿孔进行一些修饰的视网膜，并且基因治疗可以减少过滤手术

项目成果

Ohguro H, Ogawa K, Maeda T, Maruyama I, Maeda A, Takano Y, Nakazawa M: "Retinal dysfunction in cancer-associated retinopathy is improved by Ca2+ anatagonist administration and dark adaptation"Investigative Ophthalmology & Visual Science. 41(10). 2589-2595

Ohguro H、Okawa K、Maeda T、Maruyama I、Maeda A、Takano Y、Nakazawa M：“通过 Ca2 拮抗剂给药和暗适应可改善癌症相关视网膜病变中的视网膜功能障碍”调查眼科

Tanimoto N, et al.: "Electroretinographic findings in three family members with X-linked juvenile retinoschisis associated with a novel Pro192Thr mutation of the XLRS1 gene"Japanese Journal of Ophthalmology. 46 (5). 566-576 (2002)

Tanimoto N 等人：“三个患有 X 连锁青少年视网膜劈裂症的家庭成员的视网膜电图检查结果与 XLRS1 基因的新型 Pro192Thr 突变相关”，《日本眼科杂志》。

中沢 満: "眼科診療Q&A31"加齢黄斑変性の原因遺伝子とその臨床応用の可能性(分担). 1000 (2002)

Mitsuru Nakazawa：“眼科问答31”负责年龄相关性黄斑变性的基因及其临床应用的可能性（共享）1000（2002）。

Sekiya K, et al.: "Long-term fundus changes of fundus albipunctatus associated with mutations of the RDH5 gene"Arch Ophthalmol. in press.

Sekiya K 等人：“与 RDH5 基因突变相关的眼底白斑的长期眼底变化”Arch Ophasemol。

Yanagihashi S, et al.: "Autosomal dominant central areolar choroidal dystrophy and a novel Arg195Leu mutation in the peripherin/RDS gene"Arch Ophthalmol. in press.

Yanagihashi S 等人：“常染色体显性中央乳晕脉络膜营养不良和外周蛋白/RDS 基因中的新型 Arg195Leu 突变”Arch Ophasemol。