The influence of chirality on the behavior of oligonucleotides inside cells: revealing the potent cytotoxicity of G-rich l-RNA.
The influence of chirality on the behavior of oligonucleotides inside cells: revealing the potent cytotoxicity of G-rich l-RNA.
复制标题
DOI：
10.1039/d2sc05511b
复制
发表时间：
2023-02-01
期刊：
影响因子：
8.4
通讯作者：
中科院分区：
文献类型：
--
作者：
镜像l-寡核苷酸由于其固有的核酸酶抗性，越来越多地应用于生物医学研究工具和治疗方法的开发。然而，手性对生物系统中寡核苷酸行为的影响，特别是l-寡核苷酸与内源性生物大分子相互作用的程度以及由此产生的后果仍然未知。在这项研究中，我们首次表征了l-寡核苷酸的细胞内行为，揭示了寡核苷酸细胞毒性的重要手性依赖作用。我们发现外源性递送的l-寡核苷酸具有高度细胞毒性的潜力，这取决于主链化学、序列和结构。值得注意的是，对于测试的序列，我们发现单链富含g的l-RNA比它们的d-DNA/RNA对应物更具细胞毒性，具有低纳摩尔EC50值。重要的是，差异表达基因的RNA-seq分析表明，富含g的l- rna刺激先天免疫反应和促炎细胞因子的产生。这些数据不仅挑战了镜像l-寡核苷酸无毒和无免疫原性的普遍看法，而且还揭示了以前未被认识到的治疗机会。此外，通过建立序列/结构毒性关系，这项工作将指导未来如何设计和应用基于l-寡核苷酸的生物技术。我们确定了手性对人类细胞内寡核苷酸行为的影响，揭示镜像l-寡核苷酸具有高度细胞毒性的潜力。
Due to their intrinsic nuclease resistance, mirror image l-oligonucleotides are being increasingly employed in the development of biomedical research tools and therapeutics. Yet, the influence of chirality on the behavior of oligonucleotides in living systems, and specifically, the extent to which l-oligonucleotides interact with endogenous biomacromolecules and the resulting consequences remain unknown. In this study, we characterized the intracellular behavior of l-oligonucleotides for the first time, revealing important chirality-dependent effects on oligonucleotide cytotoxicity. We show that exogenously delivered l-oligonucleotides have the potential to be highly cytotoxic, which is dependent on backbone chemistry, sequence, and structure. Notably, for the sequences tested, we found that single-stranded G-rich l-RNAs are more cytotoxic than their d-DNA/RNA counterparts, exhibiting low nanomolar EC50 values. Importantly, RNA-seq analysis of differentially expressed genes suggests that G-rich l-RNAs stimulate an innate immune response and pro-inflammatory cytokine production. These data not only challenge the general perception that mirror image l-oligonucleotides are nontoxic and nonimmunogenic, but also reveal previously unrecognized therapeutic opportunities. Moreover, by establishing sequence/structure toxicity relationships, this work will guide how future l-oligonucleotide-based biotechnologies are designed and applied. We determined the influence of chirality on the behavior of oligonucleotides inside human cells, revealing that mirror-image l-oligonucleotides have the potential to be highly cytotoxic.
登录
查看更多内容
用于传感生物系统中金属离子的生物稳定 L-DNAzyme
影响因子：
7.4
作者：
Cui L;Peng R;Fu T;Zhang X;Wu C;Chen H;Liang H;Yang CJ;Tan W
通讯作者：
Tan W
影响因子：
39.3
作者：
Liu T;Zhang L;Joo D;Sun SC
通讯作者：
Sun SC
影响因子：
15
作者：
Kabza, Adam M.;Young, Brian E.;Sczepanski, Jonathan T.
通讯作者：
Sczepanski, Jonathan T.
影响因子：
7.3
作者：
Kawasaki T;Kawai T
通讯作者：
Kawai T
影响因子：
3.7
作者：
通讯作者：
--