一种独特的基因沉默方法，使用智能 RNA 表达装置（iRed），通过局部胸膜内注射在恶性胸膜间皮瘤中引起最小的免疫刺激。
A Unique Gene-Silencing Approach, Using an Intelligent RNA Expression Device (iRed), Results in Minimal Immune Stimulation When Given by Local Intrapleural Injection in Malignant Pleural Mesothelioma.
机构信息
Department of Pharmacokinetics and Biopharmaceutics, Institute of Biomedical Sciences, Tokushima University, 1-78-1, Sho-machi, Tokushima 770-8505, Japan.
Department of Bioorganic Chemistry, Institute of Biomedical Sciences, Tokushima University, 1-78-1, Sho-machi, Tokushima 770-8505, Japan.
出版信息
Molecules. 2020 Apr 9;25(7):1725. doi: 10.3390/molecules25071725.
BACKGROUND
We have recently introduced an intelligent RNA expression device (iRed), comprising the minimum essential components needed to transcribe short hairpin RNA (shRNA) in cells. Use of iRed efficiently produced shRNA molecules after transfection into cells and alleviated the innate immune stimulation following intravenous injection.
METHODS
To study the usefulness of iRed for local injection, the engineered iRed encoding luciferase shRNA (Luc iRed), complexed with cationic liposomes (Luc iRed/liposome-complexes), was intrapleurally injected into an orthotopic mesothelioma mouse model.
RESULTS
Luc iRed/liposome-complexes markedly suppressed the expression of a luciferase marker gene in pleurally disseminated mesothelioma cells. The suppressive efficiency was correlated with the expression level of shRNA within the mesothelioma cells. In addition, intrapleural injection of iRed/liposome-complexes did not induce IL-6 production in the pleural space and consequently in the blood compartment, although plasmid DNA (pDNA) or dsDNA (the natural construct for iRed) in the formulation did.
CONCLUSION
Local delivery of iRed could augment the in vivo gene silencing effect without eliciting pronounced innate immune stimulation. Our results might hold promise for widespread utilization of iRed as an RNAi-based therapeutic for intracelial malignant cancers.
背景
我们最近引入了一种智能 RNA 表达装置（iRed），它包含在细胞中转录短发夹 RNA（shRNA）所需的最小必需成分。使用 iRed 在转染细胞后可有效地产生 shRNA 分子，并减轻静脉注射后的固有免疫刺激。
方法
为了研究 iRed 用于局部注射的有用性，将工程化的 iRed 编码荧光素酶 shRNA（Luc iRed）与阳离子脂质体（Luc iRed/脂质体复合物）复合，然后经胸膜内注射到原位间皮瘤小鼠模型中。
结果
Luc iRed/脂质体复合物显著抑制了胸膜扩散性间皮瘤细胞中荧光素酶标记基因的表达。抑制效率与间皮瘤细胞内 shRNA 的表达水平相关。此外，尽管制剂中含有质粒 DNA（pDNA）或 dsDNA（iRed 的天然结构），但 iRed/脂质体复合物的胸膜内注射不会在胸膜腔和血液中引起 IL-6 的产生。
结论
局部递送 iRed 可以增强体内基因沉默效果，而不会引起明显的固有免疫刺激。我们的结果可能为广泛利用 iRed 作为基于 RNAi 的治疗剂治疗细胞内恶性癌症提供希望。
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