Improved Intravitreal AAV-Mediated Inner Retinal Gene Transduction after Surgical Internal Limiting Membrane Peeling in Cynomolgus Monkeys.
Improved Intravitreal AAV-Mediated Inner Retinal Gene Transduction after Surgical Internal Limiting Membrane Peeling in Cynomolgus Monkeys.
复制标题
DOI：
10.1016/j.ymthe.2016.10.008
复制
发表时间：
2017-01
期刊：
影响因子：
--
通讯作者：
Kazuhisa Takahashi;T. Igarashi;K. Miyake;Maika Kobayashi;Chiemi Yaguchi;O. Iijima;Yoshiyuki Yamazaki;Y. Katakai;N. Miyake;S. Kameya;T. Shimada;Hiroshi Takahashi;T. Okada
中科院分区：
文献类型：
--
作者：
Kazuhisa Takahashi;T. Igarashi;K. Miyake;Maika Kobayashi;Chiemi Yaguchi;O. Iijima;Yoshiyuki Yamazaki;Y. Katakai;N. Miyake;S. Kameya;T. Shimada;Hiroshi Takahashi;T. Okada
视网膜是基因治疗的理想靶点，因为它容易获得和有限的免疫反应。我们先前报道了在啮齿动物模型中玻璃体内注射腺相关病毒（AAV）载体以高效率转导内层视网膜。然而，在大型动物中，视网膜转导的效率较低，因为玻璃体和内界膜（ILM）充当了转导的屏障。为了克服食蟹猴中的这些障碍，我们在AAV载体注射之前进行玻璃体切除术（VIT）和ILM剥离。玻璃体内注射50 μL编码EGFP的三重突变的自身互补AAV血清型2载体后，分析转导效率。在对照组和VIT组中检测到很少的GFP表达，但在VIT+ILM组中，在剥离的ILM区域内检测到强烈的GFP表达。为了检测潜在的不良反应，我们使用彩色眼底照相、光学相干断层扫描和视网膜电图监测视网膜。未观察到与预处理相关的严重副作用。这些结果表明，在施用AAV载体之前进行手术ILM剥离对于非人灵长类动物视网膜的有效转导是安全和有用的，并为视网膜疾病的治疗提供治疗益处。
The retina is an ideal target for gene therapy because of its easy accessibility and limited immunological response. We previously reported that intravitreally injected adeno-associated virus (AAV) vector transduced the inner retina with high efficiency in a rodent model. In large animals, however, the efficiency of retinal transduction was low, because the vitreous and internal limiting membrane (ILM) acted as barriers to transduction. To overcome these barriers in cynomolgus monkeys, we performed vitrectomy (VIT) and ILM peeling before AAV vector injection. Following intravitreal injection of 50 μL triple-mutated self-complementary AAV serotype 2 vector encoding EGFP, transduction efficiency was analyzed. Little expression of GFP was detected in the control and VIT groups, but in the VIT+ILM group, strong GFP expression was detected within the peeled ILM area. To detect potential adverse effects, we monitored the retinas using color fundus photography, optical coherence tomography, and electroretinography. No serious side effects associated with the pretreatment were observed. These results indicate that surgical ILM peeling before AAV vector administration would be safe and useful for efficient transduction of the nonhuman primate retina and provide therapeutic benefits for the treatment of retinal diseases.