Wiley Luke A, Burnight Erin R, Drack Arlene V, Banach Bailey B, Ochoa Dalyz, Cranston Cathryn M, Madumba Robert A, East Jade S, Mullins Robert F, Stone Edwin M, Tucker Budd A
Department of Ophthalmology and Visual Sciences, Stephen A. Wynn Institute for Vision Research, Carver College of Medicine, University of Iowa , Iowa City, Iowa.
Hum Gene Ther. 2016 Oct;27(10):835-846. doi: 10.1089/hum.2016.049. Epub 2016 Jul 11.
Juvenile neuronal ceroid lipofuscinosis (JNCL) is a childhood neurodegenerative disease with early-onset, severe central vision loss. Affected children develop seizures and CNS degeneration accompanied by severe motor and cognitive deficits. There is no cure for JNCL, and patients usually die during the second or third decade of life. In this study, independent lines of induced pluripotent stem cells (iPSCs) were generated from two patients with molecularly confirmed mutations in CLN3, the gene mutated in JNCL. Clinical-grade adeno-associated adenovirus serotype 2 (AAV2) carrying the full-length coding sequence of human CLN3 was generated in a U.S. Food and Drug Administration-registered cGMP facility. AAV2-CLN3 was efficacious in restoring full-length CLN3 transcript and protein in patient-specific fibroblasts and iPSC-derived retinal neurons. When injected into the subretinal space of wild-type mice, purified AAV2-CLN3 did not show any evidence of retinal toxicity. This study provides proof-of-principle for initiation of a clinical trial using AAV-mediated gene augmentation for the treatment of children with CLN3-associated retinal degeneration.
青少年神经元蜡样脂褐质沉积症(JNCL)是一种儿童期神经退行性疾病,起病早,会导致严重的中心视力丧失。患病儿童会出现癫痫发作和中枢神经系统退化,并伴有严重的运动和认知缺陷。JNCL无法治愈,患者通常在生命的第二个或第三个十年死亡。在本研究中,从两名分子确诊为CLN3基因突变的患者(CLN3基因在JNCL中发生突变)中生成了诱导多能干细胞(iPSC)独立系。携带人CLN3全长编码序列的临床级腺相关腺病毒2型(AAV2)在美国食品药品监督管理局注册的cGMP设施中生产。AAV2-CLN3在恢复患者特异性成纤维细胞和iPSC衍生的视网膜神经元中的全长CLN3转录本和蛋白质方面是有效的。当将纯化的AAV2-CLN3注射到野生型小鼠的视网膜下间隙时,未显示出任何视网膜毒性的迹象。本研究为启动使用AAV介导的基因增强疗法治疗CLN3相关视网膜变性儿童的临床试验提供了原理证明。