Engineered ionizable lipid siRNA conjugates enhance endosomal escape but induce toxicity in vivo.
Engineered ionizable lipid siRNA conjugates enhance endosomal escape but induce toxicity in vivo.
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DOI：
10.1016/j.jconrel.2022.07.041
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发表时间：
2022-09
影响因子：
10.8
通讯作者：
Khvorova, Anastasia
中科院分区：
文献类型：
--
作者：
Biscans, Annabelle;Ly, Socheata;McHugh, Nicholas;Cooper, David A.;Khvorova, Anastasia
关键词：
脂质结合支持将小干扰RNA(SiRNAs)输送到肝外组织，扩大了siRNAs的治疗潜力，超出了肝脏的适应症。然而，在肝外组织中的siRNA沉默效率仍然低于在肝脏中的常规效果，部分原因是siRNA内化后的低内体逃逸。改善siRNA内体释放到细胞质中是提高脂结合siRNAs疗效的关键。鉴于可电离脂质在脂类纳米粒(LNP)的背景下能够增强内体逃逸，在此，我们首次报道了可电离脂质结合物对体内siRNA内体逃逸、组织分布、疗效和毒性的影响。在开发了一种将可电离脂质DLin-MC3-DMA共价连接到siRNAs的合成路线后，我们证明了DLin-MC3-DMA在不影响siRNA有效性的情况下增强了细胞培养中的内体逃逸。在小鼠中，DLin-MC3-DMA结合的siRNA与同样疏水的胆固醇结合的siRNA显示出相似的整体组织分布特征。然而，只有DLin-MC3-DMA结合的siRNAs聚集在血管间室中，这表明结合结构对血管内分布有影响。有趣的是，我们观察到在高积累DLin-MC3-DMA siRNAs的组织(>；20pmol/mg组织)中，基因表达的非特异性调节，而在siRNA积累较低的组织中，观察到有限的非特异性基因调节。这些发现表明，调节结合物的性质是改变siRNA胞内和胞内转运的一种很有前途的策略。微调结合物的性质以优化内体逃逸，同时将毒性降至最低，对于肝脏以外的治疗性siRNA应用的进展至关重要。
Lipid conjugation supports delivery of small interfering RNAs (siRNAs) to extrahepatic tissues, expanding the therapeutic potential of siRNAs beyond liver indications. However, siRNA silencing efficacy in extrahepatic tissues remains inferior to that routinely achieved in liver, partially due to the low rate of endosomal escape following siRNA internalization. Improving siRNA endosomal release into cytoplasm is crucial to improving efficacy of lipid-conjugated siRNAs. Given the ability of ionizable lipids to enhance endosomal escape in a context of lipid nanoparticles (LNP), here, we provide the first report on the effect of an ionizable lipid conjugate on siRNA endosomal escape, tissue distribution, efficacy, and toxicity in vivo. After developing a synthetic route to covalently attach the ionizable lipid, DLin-MC3-DMA, to siRNAs, we demonstrate that DLin-MC3-DMA enhances endosomal escape in cell culture without compromising siRNA efficacy. In mice, DLin-MC3-DMA conjugated siRNAs exhibit a similar overall tissue distribution profile to the similarly hydrophobic cholesterol-conjugated siRNA. However, only DLin-MC3-DMA conjugated siRNAs accumulated in vascular compartments, suggesting an effect of conjugate structure on intratissue distribution. Interestingly, we observed non-specific modulation of gene expression in tissues with high accumulation of DLin-MC3-DMA siRNAs (>20 pmol/mg of tissue) while limited non-specific gene modulation has been observed in tissues with lower siRNA accumulation. These findings suggest modulating the nature of the conjugate is a promising strategy to alter siRNA intratissue and intracellular trafficking. Fine-tuning the nature of the conjugate to optimize endosomal escape while minimizing toxicity will be critical for the progression of therapeutic siRNA applications beyond the liver.
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DOI：
10.1038/mtna.2016.55
发表时间：
2016-08-02
期刊：
Molecular therapy. Nucleic acids
影响因子：
--
作者：
通讯作者：
--
影响因子：
5.9
作者：
Munson MJ;O'Driscoll G;Silva AM;Lázaro-Ibáñez E;Gallud A;Wilson JT;Collén A;Esbjörner EK;Sabirsh A
通讯作者：
Sabirsh A
影响因子：
46.9
作者：
Gilleron, Jerome;Querbes, William;Zerial, Marino
通讯作者：
Zerial, Marino
影响因子：
158.5
作者：
Balwani, Manisha;Sardh, Eliane;Gouya, Laurent
通讯作者：
Gouya, Laurent
影响因子：
16.6
作者：
Jayaraman, Muthusamy;Ansell, Steven M.;Mui, Barbara L.;Tam, Ying K.;Chen, Jianxin;Du, Xinyao;Butler, David;Eltepu, Laxman;Matsuda, Shigeo;Narayanannair, Jayaprakash K.;Rajeev, Kallanthottathil G.;Hafez, Ismail M.;Akinc, Akin;Maier, Martin A.;Tracy, Mark A.;Cullis, Pieter R.;Madden, Thomas D.;Manoharan, Muthiah;Hope, Michael J.
通讯作者：
Hope, Michael J.