Murray Susan F, Jazayeri Ali, Matthes Michael T, Yasumura Douglas, Yang Haidong, Peralta Raechel, Watt Andy, Freier Sue, Hung Gene, Adamson Peter S, Guo Shuling, Monia Brett P, LaVail Matthew M, McCaleb Michael L
Isis Pharmaceuticals Carlsbad, California, United States.
University of California at San Francisco School of Medicine, Beckman Vision Center, San Francisco, California, United States.
Invest Ophthalmol Vis Sci. 2015 Oct;56(11):6362-75. doi: 10.1167/iovs.15-16400.
To preserve photoreceptor cell structure and function in a rodent model of retinitis pigmentosa with P23H rhodopsin by selective inhibition of the mutant rhodopsin allele using a second generation antisense oligonucleotide (ASO).
Wild-type mice and rats were treated with ASO by intravitreal (IVT) injection and rhodopsin mRNA and protein expression were measured. Transgenic rats expressing the murine P23H rhodopsin gene (P23H transgenic rat Line 1) were administered either a mouse-specific P23H ASO or a control ASO. The contralateral eye was injected with PBS and used as a comparator control. Electroretinography (ERG) measurements and analyses of the retinal outer nuclear layer were conducted and correlated with rhodopsin mRNA levels.
Rhodopsin mRNA and protein expression was reduced after a single ASO injection in wild-type mice with a rhodopsin-specific ASO. Transgenic rat eyes that express a murine P23H rhodopsin gene injected with a murine P23H ASO had a 181 ± 39% better maximum amplitude response (scotopic a-wave) as compared with contralateral PBS-injected eyes; the response in control ASO eyes was not significantly different from comparator contralateral eyes. Morphometric analysis of the outer nuclear layer showed a significantly thicker nuclear layer in eyes injected with murine P23H ASO (18%) versus contralateral PBS-injected eyes.
Allele-specific ASO-mediated knockdown of mutant P23H rhodopsin expression slowed the rate of photoreceptor degeneration and preserved the function of photoreceptor cells in eyes of the P23H rhodopsin transgenic rat. Our data indicate that ASO treatment is a potentially effective therapy for the treatment of retinitis pigmentosa.
通过使用第二代反义寡核苷酸（ASO）选择性抑制突变型视紫红质等位基因，在携带P23H视紫红质的视网膜色素变性啮齿动物模型中保留光感受器细胞结构和功能。
通过玻璃体内（IVT）注射用ASO处理野生型小鼠和大鼠，并测量视紫红质mRNA和蛋白质表达。给表达鼠P23H视紫红质基因的转基因大鼠（P23H转基因大鼠品系1）注射小鼠特异性P23H ASO或对照ASO。对侧眼注射磷酸盐缓冲盐水（PBS）并用作对照。进行视网膜电图（ERG）测量和视网膜外核层分析，并与视紫红质mRNA水平相关联。
用视紫红质特异性ASO单次注射后，野生型小鼠的视紫红质mRNA和蛋白质表达降低。注射鼠P23H ASO的表达鼠P23H视紫红质基因的转基因大鼠眼与对侧注射PBS的眼相比，最大振幅反应（暗视a波）改善了181±39％；对照ASO眼的反应与对侧对照眼无显著差异。外核层的形态计量分析显示，注射鼠P23H ASO的眼的核层（18％）比注射对侧PBS的眼明显更厚。
等位基因特异性ASO介导的突变型P23H视紫红质表达的敲低减缓了光感受器变性速率，并保留了P23H视紫红质转基因大鼠眼中光感受器细胞的功能。我们的数据表明，ASO治疗是治疗视网膜色素变性的一种潜在有效疗法。
