Engineered Histidine-Enriched Facial Lipopeptides for Enhanced Intracellular Delivery of Functional siRNA to Triple Negative Breast Cancer Cells
Engineered Histidine-Enriched Facial Lipopeptides for Enhanced Intracellular Delivery of Functional siRNA to Triple Negative Breast Cancer Cells
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DOI：
10.1021/acsami.8b13794
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发表时间：
2019-02-06
影响因子：
9.5
通讯作者：
Roy, Rituparna Sinha
中科院分区：
文献类型：
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作者：
Biswas, Abhijit;Chakraborty, Kasturee;Roy, Rituparna Sinha
功能性 siRNA 的胞质递送仍然是开发基于 siRNA 的疗法的主要挑战。设计临床安全有效的 siRNA 转运蛋白以跨质膜和内体膜递送功能性 siRNA 仍然是一个关键障碍。为了改善内体释放，我们设计了具有 Arg-(D)His-Arg 模板的环状和线性肽转运蛋白。计算研究表明，Arg-(D)His-Arg 模板在生理 pH 下也通过 Arg-His 侧链氢键相互作用稳定，在较低 pH 下解离。通过分子动力学模拟检查了整体原子相互作用，这表明肽_siRNA组装形成的程度很大程度上取决于肽的物理化学性质。我们设计的肽具有 Arg-(D)His-Arg 模板和两个脂质部分，有助于高产率地进行 siRNA 的细胞内递送。此外，引入不饱和脂质、亚油酸部分以促进融合性并促进内体释放和胞质递送。有趣的是，这种蛋白酶抗性肽为 siRNA 提供了血清稳定性，并在三阴性乳腺癌 (TNBC) 细胞系中表现出高效的 erk1 和 erk2 基因沉默。 erk1 和 erk2 基因敲低实验证明，具有两个亚油基部分的肽与商业转染试剂 HiPerFect 具有相当的功效。此外，我们的研究表明，对于 TNBC 治疗，ERK1/2 沉默 siRNA 和负载阿霉素的短杆菌肽介导的联合疗法比 siRNA 介导的基因沉默单一疗法更有效。
Cytosolic delivery of functional siRNA remains the major challenge to develop siRNA-based therapeutics. Designing clinically safe and effective siRNA transporter to deliver functional siRNA across the plasma and endosomal membrane remains a key hurdle. With the aim of improving endosomal release, we have designed cyclic and linear peptide-based transporters having an Arg-(D)His-Arg template. Computational studies show that the Arg-(D)His-Arg template is also stabilized by the Arg-His side-chain hydrogen bonding interaction at physiological pH, which dissociates at lower pH. The overall atomistic interactions were examined by molecular dynamics simulations, which indicate that the extent of peptide_siRNA assembly formation depends greatly on physicochemical properties of the peptides. Our designed peptides having the Arg-(D)His-Arg template and two lipidic moieties facilitate high yield of intracellular delivery of siRNA. Additionally, unsaturated lipid, linoleic acid moieties were introduced to promote fusogenicity and facilitate endosomal release and cytosolic delivery. Interestingly, such protease-resistant peptides provide serum stability to siRNA and exhibit high efficacy of erk1 and erk2 gene silencing in the triple negative breast cancer (TNBC) cell line. The peptide having two linoleyl moieties demonstrated comparable efficacy with commercial transfection reagent HiPerFect, as evidenced by the erk1 and erk2 gene knockdown experiment. Additionally, our study shows that ERK1/2 silencing siRNA and doxorubicin-loaded gramicidin-mediated combination therapy is more effective than siRNA-mediated gene silencing-based monotherapy for TNBC treatment.