Department of Chemical and Biological Engineering, University of Sheffield, S1 3JD, UK.
Centre for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266555, China.
Int J Pharm. 2022 Apr 5;617:121619. doi: 10.1016/j.ijpharm.2022.121619. Epub 2022 Feb 23.
Gene therapy has gained increasing attention as an alternative to pharmacotherapy for treatment of various diseases. The extracellular and intracellular barriers to gene delivery necessitate the use of gene vectors which has led to the development of myriads of gene delivery systems. However, many of these gene delivery systems have pitfalls such as low biocompatibility, low loading efficiency, low transfection efficiency, lack of tissue selectivity and high production costs. Herein, we report the development of a new series of short cationic amphiphilic peptides with anticancer activity for selective delivery of small interfering RNA (siRNA) and antisense oligodeoxynucleotides (ODNs) to cancer cells. The peptides consist of alternating dyads of hydrophobic (isoleucine (I) or leucine (L)) and hydrophilic (arginine (R) or lysine (L)) amino acids. The peptides exhibited higher preference for transfection of HCT 116 colorectal cancer cells compared to human dermal fibroblasts (HDFs) and induced higher level of gene silencing in the cancer cells. The nucleic acid complexation and transfection efficiency of the peptides was a function of their secondary structure, their hydrophobicity and their C-terminal amino acid. The peptides containing L in their hydrophobic domain formed stronger complexes with siRNA and successfully delivered it to the cancer cells but were unable to release their cargo inside the cells and therefore could not induce any gene silencing. On the contrary, the peptides containing I in their hydrophobic domain were able to release their associated siRNA and induce considerable gene silencing in cancer cells. The peptides exhibited higher selectivity for colorectal cancer cells and induced less gene silencing in fibroblasts compared to the lipid-based commercial transfection reagent DharmaFECT™ 1. The results from this study can serve as a tool for rational design of new peptide-based gene vectors for high selective gene delivery to cancer cells.
基因治疗作为一种替代药物治疗各种疾病的方法，越来越受到关注。基因传递的细胞外和细胞内屏障需要使用基因载体，这导致了无数基因传递系统的发展。然而，许多这些基因传递系统存在缺陷，如低生物相容性、低装载效率、低转染效率、缺乏组织选择性和高生产成本。在这里，我们报告了一系列具有抗癌活性的新型短阳离子两亲肽的开发，用于选择性地将小干扰 RNA(siRNA)和反义寡脱氧核苷酸(ODNs)递送到癌细胞。这些肽由疏水性(异亮氨酸(I)或亮氨酸(L))和亲水性(精氨酸(R)或赖氨酸(L))氨基酸交替组成的二联体组成。与人类真皮成纤维细胞(HDFs)相比，这些肽对结直肠癌细胞(HCT 116)的转染具有更高的偏好性，并在癌细胞中诱导更高水平的基因沉默。肽的核酸复合物形成和转染效率是其二级结构、疏水性和 C 末端氨基酸的函数。其疏水性结构域中含有 L 的肽与 siRNA 形成更强的复合物，并成功地将其递送到癌细胞中，但无法在细胞内释放其货物，因此不能诱导任何基因沉默。相反，其疏水性结构域中含有 I 的肽能够释放其相关的 siRNA，并在癌细胞中诱导相当程度的基因沉默。与基于脂质的商业转染试剂 DharmaFECT™1 相比，这些肽对结直肠癌细胞具有更高的选择性，并在成纤维细胞中诱导较少的基因沉默。本研究的结果可作为合理设计新型基于肽的基因载体以高选择性递送至癌细胞的工具。
