Allele-Specific Inhibition of Rhodopsin With an Antisense Oligonucleotide Slows Photoreceptor Cell Degeneration
Allele-Specific Inhibition of Rhodopsin With an Antisense Oligonucleotide Slows Photoreceptor Cell Degeneration
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DOI：
10.1167/iovs.15-16400
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发表时间：
2015-10-01
影响因子：
4.4
通讯作者：
McCaleb, Michael L.
中科院分区：
文献类型：
--
作者：
Murray, Susan F.;Jazayeri, Ali;McCaleb, Michael L.
目的.目的：通过第二代反义寡核苷酸（阿索）选择性抑制P23 H视紫红质突变型视紫红质等位基因，保护视网膜色素变性（retinitis pigmentosa）啮齿动物模型的感光细胞结构和功能。通过玻璃体内（IVT）注射用阿索处理野生型小鼠和大鼠，并测量视紫红质mRNA和蛋白质表达。向表达鼠P23 H视紫红质基因的转基因大鼠（P23 H转基因大鼠品系1）施用小鼠特异性P23 H阿索或对照阿索。对侧眼注射PBS并用作比较对照。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。野生型小鼠单次注射阿索后，视紫红质mRNA和蛋白质表达减少，注射了视紫红质特异性阿索。与注射PBS的对侧眼睛相比，注射鼠P23 H阿索的表达鼠P23 H视紫红质基因的转基因大鼠眼睛具有181 +/-39%的最大振幅反应（暗视a波）;对照阿索眼睛中的反应与比较对侧眼睛没有显著差异。外核层的形态测定分析显示，与对侧PBS注射的眼睛相比，注射鼠P23 H阿索的眼睛（18%）的核层显著更厚。结论.等位基因特异性ASO介导的突变型P23 H视紫红质表达的敲低减缓了P23 H视紫红质转基因大鼠眼睛中感光细胞变性的速率并保留了感光细胞的功能。我们的数据表明，阿索治疗是一种潜在的有效治疗视网膜色素变性。
PURPOSE. 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).METHODS. 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.RESULTS. 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. CONCLUSIONS. 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.