长效 RNA 干扰基因治疗在显性视网膜色素变性小鼠模型中的应用。
Long-term RNA interference gene therapy in a dominant retinitis pigmentosa mouse model.
机构信息
Department of Ophthalmology, University of Utah Health Science Center, Salt Lake City, UT 84132, USA.
出版信息
Proc Natl Acad Sci U S A. 2011 Nov 8;108(45):18476-81. doi: 10.1073/pnas.1112758108. Epub 2011 Oct 31.
Abstract
RNA interference (RNAi) gene silencing is a potential therapeutic strategy for dominant retinal degeneration disorders. We used self-complementary (sc) AAV2/8 vector to develop an RNAi-based gene therapy in a dominant retinal degeneration mouse model expressing bovine GCAP1(Y99C). We established an in vitro shRNA screening assay based on EGFP-tagged bovine GCAP1, and identified a shRNA that effectively silenced the bovine GCAP1 transgene with ∼80% efficiency. Subretinal injection of scAAV2/8 carrying shRNA expression cassette showed robust expression as early as 1 wk after injection. The gene silencing significantly improved photoreceptor survival, delayed disease onset, and increased visual function. Our results provide a promising strategy toward effective RNAi-based gene therapy by scAAV2/8 delivery for dominant retinal diseases.
摘要
RNA 干扰 (RNAi) 基因沉默是治疗显性视网膜退行性疾病的一种有潜力的治疗策略。我们使用自我互补 (sc) AAV2/8 载体，在表达牛 GCAP1(Y99C)的显性视网膜退行性小鼠模型中开发了一种基于 RNAi 的基因治疗方法。我们建立了一种基于 EGFP 标记的牛 GCAP1 的体外 shRNA 筛选检测方法，并鉴定出一种能够有效沉默牛 GCAP1 转基因的 shRNA，效率约为 80%。携带 shRNA 表达盒的 scAAV2/8 经视网膜下注射后，在注射后 1 周即可观察到强烈的表达。基因沉默显著提高了感光细胞的存活率，延缓了疾病的发作，并增强了视觉功能。我们的研究结果为通过 scAAV2/8 递送来实现有效的基于 RNAi 的显性视网膜疾病基因治疗提供了一种很有前途的策略。
相似文献
1
Long-term RNA interference gene therapy in a dominant retinitis pigmentosa mouse model.长效 RNA 干扰基因治疗在显性视网膜色素变性小鼠模型中的应用。
Proc Natl Acad Sci U S A. 2011 Nov 8;108(45):18476-81. doi: 10.1073/pnas.1112758108. Epub 2011 Oct 31.
2
RNA interference gene therapy in dominant retinitis pigmentosa and cone-rod dystrophy mouse models caused by GCAP1 mutations.RNA干扰基因疗法在由GCAP1突变引起的显性视网膜色素变性和视锥-视杆营养不良小鼠模型中的应用
Front Mol Neurosci. 2014 Apr 7;7:25. doi: 10.3389/fnmol.2014.00025. eCollection 2014.
3
RNAi-mediated gene suppression in a GCAP1(L151F) cone-rod dystrophy mouse model.RNAi 介导的 GCAP1(L151F) 型 Cone-Rod 营养不良小鼠模型中的基因抑制。
PLoS One. 2013;8(3):e57676. doi: 10.1371/journal.pone.0057676. Epub 2013 Mar 5.
4
Mutation-independent rescue of a novel mouse model of Retinitis Pigmentosa.突变非依赖性挽救新型视网膜色素变性小鼠模型。
Gene Ther. 2013 Apr;20(4):425-34. doi: 10.1038/gt.2012.53. Epub 2012 Jul 19.
5
Development of gene therapy for treatment of age-related macular degeneration.用于治疗年龄相关性黄斑变性的基因疗法的发展。
Acta Ophthalmol. 2014 Jul;92 Thesis3:1-38. doi: 10.1111/aos.12452.
6
GUCY2D Cone-Rod Dystrophy-6 Is a "Phototransduction Disease" Triggered by Abnormal Calcium Feedback on Retinal Membrane Guanylyl Cyclase 1.GUCY2D Cone-Rod 营养不良症-6 是一种“光转导疾病”，由视网膜膜鸟苷酸环化酶 1 上异常钙反馈引发。
J Neurosci. 2018 Mar 21;38(12):2990-3000. doi: 10.1523/JNEUROSCI.2985-17.2018. Epub 2018 Feb 12.
7
Improved retinal function in a mouse model of dominant retinitis pigmentosa following AAV-delivered gene therapy.在腺相关病毒介导的基因治疗后，显性视网膜色素变性小鼠模型的视网膜功能得到改善。
Mol Ther. 2009 Apr;17(4):593-9. doi: 10.1038/mt.2008.301. Epub 2009 Jan 27.
8
Repair of rhodopsin mRNA by spliceosome-mediated RNA trans-splicing: a new approach for autosomal dominant retinitis pigmentosa.通过剪接体介导的RNA反式剪接修复视紫红质mRNA：常染色体显性遗传性视网膜色素变性的一种新方法。
Mol Ther. 2015 May;23(5):918-930. doi: 10.1038/mt.2015.11. Epub 2015 Jan 26.
9
Gene therapy in animal models of autosomal dominant retinitis pigmentosa.常染色体显性遗传性视网膜色素变性动物模型中的基因治疗
Mol Vis. 2012;18:2479-96. Epub 2012 Oct 6.
10
Genomic form of rhodopsin DNA nanoparticles rescued autosomal dominant Retinitis pigmentosa in the P23H knock-in mouse model.载脂蛋白 E 基因多态性与阿尔茨海默病的相关性：一项荟萃分析和系统综述
Biomaterials. 2018 Mar;157:26-39. doi: 10.1016/j.biomaterials.2017.12.004. Epub 2017 Dec 5.
引用本文的文献
1
Novel compound heterozygous mutation associated with retinal cone dystrophy.与视网膜锥体细胞营养不良相关的新型复合杂合突变。
Exp Ther Med. 2025 Jun 10;30(2):155. doi: 10.3892/etm.2025.12905. eCollection 2025 Aug.
2
Next generation therapeutics for retinal neurodegenerative diseases.视网膜神经退行性疾病的下一代治疗方法。
J Control Release. 2024 Mar;367:708-736. doi: 10.1016/j.jconrel.2024.01.063. Epub 2024 Feb 10.
3
CRISPR/SaCas9-based gene editing rescues photoreceptor degeneration throughout a rhodopsin-associated autosomal dominant retinitis pigmentosa mouse model.基于 CRISPR/SaCas9 的基因编辑挽救了整个视紫红质相关常染色体显性遗传性视网膜色素变性小鼠模型中的光感受器变性。
Exp Biol Med (Maywood). 2023 Oct;248(20):1818-1828. doi: 10.1177/15353702231199069. Epub 2023 Oct 14.
4
Gene therapy and therapeutic editing with outer or inner retina animal models.利用外视网膜或内视网膜动物模型进行基因治疗和治疗性编辑。
Vision Res. 2023 Dec;213:108316. doi: 10.1016/j.visres.2023.108316. Epub 2023 Sep 15.
5
Rhodopsin-associated retinal dystrophy: Disease mechanisms and therapeutic strategies.视紫红质相关视网膜营养不良：疾病机制与治疗策略。
Front Neurosci. 2023 Apr 3;17:1132179. doi: 10.3389/fnins.2023.1132179. eCollection 2023.
6
-Related Cone-Rod Dystrophy: Clinical Characteristics, Imaging Findings, and Genetic Test Results-A Case Report.相关 Cone-Rod 营养不良：临床特征、影像学发现和基因检测结果——病例报告。
Medicina (Kaunas). 2023 Feb 17;59(2):399. doi: 10.3390/medicina59020399.
7
Characterization of two novel knock-in mouse models of syndromic retinal ciliopathy carrying hypomorphic mutations.两种新型综合征性视网膜纤毛病 knock-in 小鼠模型的特征，这些模型携带有功能丧失的突变。
Zool Res. 2022 May 18;43(3):442-456. doi: 10.24272/j.issn.2095-8137.2021.387.
8
Systemic administration of AAV-Slc25a46 mitigates mitochondrial neuropathy in Slc25a46-/- mice.AAV-Slc25a46 全身给药可减轻 Slc25a46-/- 小鼠的线粒体神经病变。
Hum Mol Genet. 2020 Mar 13;29(4):649-661. doi: 10.1093/hmg/ddz277.
9
Allele-Specific CRISPR-Cas9 Genome Editing of the Single-Base P23H Mutation for Rhodopsin-Associated Dominant Retinitis Pigmentosa.针对视紫红质相关显性视网膜色素变性的单碱基P23H突变进行等位基因特异性CRISPR-Cas9基因组编辑
CRISPR J. 2018 Feb;1(1):55-64. doi: 10.1089/crispr.2017.0009.
10
Progressive cone and cone-rod dystrophies: clinical features, molecular genetics and prospects for therapy.进行性视锥细胞和视锥-视杆细胞营养不良：临床特征、分子遗传学及治疗前景
Br J Ophthalmol. 2019 Jan 24;103(5):711-20. doi: 10.1136/bjophthalmol-2018-313278.
本文引用的文献
1
Bevasiranib for the treatment of wet, age-related macular degeneration.贝伐西单抗用于治疗湿性年龄相关性黄斑变性。
Ophthalmol Eye Dis. 2010 Dec 19;2:75-83. doi: 10.4137/OED.S4878. Print 2010.
2
Long-term preservation of cone photoreceptors and restoration of cone function by gene therapy in the guanylate cyclase-1 knockout (GC1KO) mouse.通过基因治疗在鸟苷酸环化酶-1 敲除（GC1KO）小鼠中对锥形光感受器进行长期保存和恢复锥形功能。
Invest Ophthalmol Vis Sci. 2011 Sep 9;52(10):7098-108. doi: 10.1167/iovs.11-7867.
3
Role of guanylyl cyclase modulation in mouse cone phototransduction.环鸟苷酸调节在小鼠视锥光传导中的作用。
J Neurosci. 2011 Jun 1;31(22):7991-8000. doi: 10.1523/JNEUROSCI.6650-10.2011.
4
RNAi: a potential new class of therapeutic for human genetic disease.RNAi：一种治疗人类遗传疾病的潜在新型治疗药物。
Hum Genet. 2011 Nov;130(5):583-605. doi: 10.1007/s00439-011-0995-8. Epub 2011 May 3.
5
Current prospects for RNA interference-based therapies.基于 RNA 干扰的疗法的当前前景。
Nat Rev Genet. 2011 May;12(5):329-40. doi: 10.1038/nrg2968.
6
Suppression and replacement gene therapy for autosomal dominant disease in a murine model of dominant retinitis pigmentosa.抑制和替代基因疗法治疗显性视网膜炎色素变性的常染色体显性疾病的小鼠模型。
Mol Ther. 2011 Apr;19(4):642-9. doi: 10.1038/mt.2010.293. Epub 2011 Jan 11.
7
Recognizing and avoiding siRNA off-target effects for target identification and therapeutic application.鉴定和避免 siRNA 脱靶效应，以进行靶标鉴定和治疗应用。
Nat Rev Drug Discov. 2010 Jan;9(1):57-67. doi: 10.1038/nrd3010.
8
Increased light exposure alleviates one form of photoreceptor degeneration marked by elevated calcium in the dark.增加光照可缓解一种感光细胞退化的形式，这种退化的特征是在黑暗中钙含量升高。
PLoS One. 2009 Dec 24;4(12):e8438. doi: 10.1371/journal.pone.0008438.
9
AAV-mediated knockdown of peripherin-2 in vivo using miRNA-based hairpins.利用基于 miRNA 的发夹结构在体内介导外周蛋白-2 的 AAV 敲低。
Gene Ther. 2010 Apr;17(4):486-93. doi: 10.1038/gt.2009.162. Epub 2009 Dec 10.
10
Adeno-associated virus-mediated rhodopsin replacement provides therapeutic benefit in mice with a targeted disruption of the rhodopsin gene.腺相关病毒介导的视蛋白替代为视蛋白基因靶向敲除的小鼠提供了治疗益处。
Hum Gene Ther. 2010 Mar;21(3):311-23. doi: 10.1089/hum.2009.119.
Zotero 插件