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复杂性，被提名为日本成年人口中法律失明的第三个最常见原因，其中有3500人中有1人有1人，因此，就反对失明政策而言，这是一种重要疾病。目前，世界上许多研究人员都研究了基因疗法，视网膜移植和视觉假体，并有望作为有效治疗发展。在这项研究中，我们研究了通过电穿孔转移到视网膜的技术可能性和影响，作为一种残留变性的基因治疗方法。为了测试技术的可能性，我们在兔眼上使用了PARS植物玻璃体切除术，接近视网膜，在视网膜下空间注入DNA溶液，并用针电极施加电穿孔。结果表明，该方法只能在视网膜的一小部分中传递DNA，这可能不足以引起……更多的久远效应。这也表明我们需要开发另一种类型的电极，例如杯形。作为研究的第二部分，我们研究了电基因转移到眼组织的表达和影响。首先，我们利用结膜组织作为视网膜前基因转移的靶标，因为将DNA转移到结膜上比视网膜更容易。表明，通过电穿孔转移到结膜的绿色荧光蛋白显然在转移后30天在目标组织中表达。然后，我们试图将金属蛋白酶3 cDNA转移到小梁切除术期间兔眼上的结膜皮瓣，以检查术后基因对眼内压的作用。结果表明，用基因转移治疗的眼睛的眼内压与小梁切除术用MMC处理的眼睛一样低，并且在注射DNA的区域没有病理反应。所有这些结果都表明，我们在视网膜之前成功将DNA片段转移到结膜上，因此我们有可能使用一些修饰的电穿孔将基因转移到视网膜上，并且该基因治疗可以应用于格拉糖果滤过手术。较少的

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