Drug Design, Development and Therapy (Apr 2019)
Transportan-derived cell-penetrating peptide delivers siRNA to inhibit replication of influenza virus in vivo
Abstract
Cuiling Zhang,1 Weigang Ren,1 Qingxin Liu,2 Zhikai Tan,3 Junwei Li,1 Chunyi Tong3 1College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, People’s Republic of China; 2Jiangsu Vocational College of Agriculture and Forestry, Jurong 212400, People’s Republic of China; 3Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biology, Hunan University, Changsha 410082, People’s Republic of China Introduction: In this study, we report on the development of an effective delivery system for siRNAs; a novel cell-penetrating peptide (CPP), T9(dR), obtained from transportan (TP), was used for in vivo and in vitro testing. Methods: In this study, toxicity of T9(dR) and TP and efficient delivery of siRNA were tested in 293T, MDCK, RAW, and A549 cells. Furthermore, T9(dR)- and TP-delivered siRNAs against nucleoprotein (NP) gene segment of influenza virus (siNP) were studied in both cell lines and mice. Results: Gel retardation showed that T9(dR) effectively condensed siRNA into nanoparticles sized between 350 and 550 nm when the mole ratio of T9(dR) to siRNA was ≥4:1. In vitro studies demonstrated that T9(dR) successfully delivered siRNA with low cellular toxicity into several cell lines. It was also observed that T9(dR)-delivered siRNAs inhibited replication of influenza virus more efficiently as compared to that delivered by TP into the MDCK and A549 cells. It was also noticed that when given a combined tail vein injection of siNP and T9(dR) or TP, all mice in the 50 nmol siNP group infected with PR8 influenza virus survived and showed weight recovery at 2 weeks post-infection. Conclusion: This study indicates that T9(dR) is a promising siRNA delivery tool with potential application for nucleotide drug delivery. Keywords: cell-penetrating peptide, CPP, siRNA, inhibition, influenza virus, IV, transportan, nucleoprotein, NP
