Gene Therapy in the Rd6 Mouse Model of Retinal Degeneration
Gene Therapy in the Rd6 Mouse Model of Retinal Degeneration
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DOI：
10.1007/978-1-4614-3209-8_89
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发表时间：
2014-01-01
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影响因子：
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通讯作者：
Hauswirth, William W.
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文献类型：
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作者：
Dinculescu, Astra;Min, Seok-Hong;Hauswirth, William W.
rd6小鼠是由Mfrp（膜型卷曲相关蛋白）基因突变引起的基于rpe（视网膜色素上皮）的常染色体隐性视网膜色素变性（RP）的天然模型。在此之前，我们发现由酪氨酸衣壳突变体scAAV8 （Y733F）载体介导的野生型小鼠Mfrp视网膜下递送可防止光受体细胞死亡，并通过视网膜电图评估恢复视网膜功能。在这项研究中，我们描述了基因治疗对rd6小鼠视网膜结构和功能的影响，使用四重（Y272, 444,500,730f）酪氨酸衣壳突变scAAV2病毒载体在出生后14天（P14）通过视网膜下传递。我们表明，治疗是有效的减缓光感受器变性，并防止在视网膜下空间异常吞噬细胞的特征性积累。在处理过的rd6小鼠中，由普遍存在的鸡β -肌动蛋白（smCBA）启动子驱动的MFRP表达主要存在于RPE顶端膜及其微绒毛的整个长度，以及光感受器内段，这表明可能与肌动蛋白丝相互作用。尽管保留了视网膜形态，但基因治疗对视网膜功能的影响很小，这表明scAAV8 （Y733F）载体可能更有效地治疗由Mfrp突变引起的RP。
The rd6 mouse is a natural model of an RPE-based (retinal pigment epithelium) autosomal recessive retinitis pigmentosa (RP) caused by mutations in the Mfrp (membrane-type frizzled related protein) gene. Previously, we showed that subretinal delivery of the wild-type mouse Mfrp mediated by a tyrosine-capsid mutant scAAV8 (Y733F) vector prevented photoreceptor cell death, and rescued retinal function as assessed by electroretinography. In this study, we describe the effect of gene therapy on the retinal structure and function in rd6 mice using a quadruple (Y272, 444, 500, 730F) tyrosine-capsid mutant scAAV2 viral vector delivered subretinally at postnatal day 14 (P14). We show that therapy is effective at slowing the photoreceptor degeneration, and in preventing the characteristic accumulation of abnormal phagocytic cells in the subretinal space. MFRP expression as driven by the ubiquitous chicken beta-actin (smCBA) promoter in treated rd6 mice was found predominantly in the RPE apical membrane and the entire length of its microvilli, as well as in the photoreceptor inner segments, suggesting a potential interaction with actin filaments. In spite of preserving retinal morphology, the effects of gene therapy on retinal function were minimal, suggesting that the scAAV8 (Y733F) vector may be more efficient for the treatment of RP caused by Mfrp mutations.