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Delivery of Antisense Oligonucleotides to the Cornea.
Nucleic Acid Therapeutics ( IF 4 ) Pub Date : 2020-08-06 , DOI: 10.1089/nat.2019.0838
Viet Q Chau 1 , Jiaxin Hu 2 , Xin Gong 1 , John D Hulleman 1 , Rafael L Ufret-Vincenty 1 , Frank Rigo 3 , Thahza P Prakash 3 , David R Corey 2 , V Vinod Mootha 1, 4
Affiliation  

Antisense oligonucleotides (ASOs) are synthetic nucleic acids that recognize complementary RNA sequences inside cells and modulate gene expression. In this study, we explore the feasibility of ASO delivery to the cornea. We used quantitative polymerase chain reaction to test the efficacy of a benchmark ASO targeting a noncoding nuclear RNA, Metastasis-Associated Lung Adenocarcinoma Transcript 1 (MALAT1), in a human corneal endothelial cell line, ex vivo human corneas, and in vivo in mice. In vivo delivery was via intravitreal or intracameral injections as well as topical administration. The anti-MALAT1 ASO significantly reduced expression of MALAT1 in a corneal endothelial cell line. We achieved a dose-dependent reduction of target gene expression in endothelial tissue from ex vivo human donor corneas. In vivo mouse experiments confirmed MALAT1 reduction in whole corneal tissue via intravitreal and intracameral routes, 82% and 71% knockdown, respectively (P < 0.001). Effects persisted up to at least 21 days, 32% (P < 0.05) and 43% (P < 0.05) knockdown, respectively. We developed protocols for the isolation and analysis of mouse corneal endothelium and observed reduction in MALAT1 expression upon both intravitreal and intracameral administrations, 64% (P < 0.05) and 63% (P < 0.05) knockdown, respectively. These data open the possibility of using ASOs to treat corneal disease.

中文翻译:

将反义寡核苷酸递送至角膜。

反义寡核苷酸 (ASO) 是合成核酸,可识别细胞内的互补 RNA 序列并调节基因表达。在这项研究中,我们探讨了 ASO 递送至角膜的可行性。我们使用定量聚合酶链反应来测试针对非编码核 RNA 转移相关肺腺癌转录本 1 ( MALAT1 )的基准 ASO在人角膜内皮细胞系、离体人角膜和小鼠体内的功效。体内递送是通过玻璃体内或前房内注射以及局部给药。抗MALAT1 ASO 显着降低MALAT1 的表达在角膜内皮细胞系中。我们实现了离体人类供体角膜内皮组织中靶基因表达的剂量依赖性降低。体内小鼠实验证实,通过玻璃体内和前房内途径,整个角膜组织中的MALAT1减少,分别为 82% 和 71% ( P  < 0.001)。效果持续至少 21 天,分别为 32% ( P  < 0.05) 和 43% ( P  < 0.05) 击倒。我们开发了用于分离和分析小鼠角膜内皮的方案,并观察到玻璃体内和前房内给药后MALAT1表达降低,分别为 64% ( P  < 0.05) 和 63% (P  < 0.05) 分别击倒。这些数据开启了使用 ASO 治疗角膜疾病的可能性。
更新日期:2020-08-12
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