Design and evaluation of endosomolytic biocompatible peptides as carriers for siRNA delivery.
Gene therapy using RNA interference (RNAi) technology has been explored to treat cancers, by regulating the expression of oncogene. However, even though small interfering RNA (siRNA), which triggers RNAi, may have great therapeutic potential, efforts at using them in vivo have been hampered by the difficulty of effective and safe delivery into cells of interest. In this study, to develop a safe and efficient carrier for in vitro and in vivo siRNA delivery, we designed a peptide library. These peptides are improved variants of a known peptide based siRNA carrier C6. All the modifications improved the transfection efficiency of C6 to some degree. After completing prescreening for activity, several promising candidates were used for further evaluation. Selected peptides C6M3 and C6M6 could form stable complexes with siRNA. These complexes could be greatly uptaken by cells and showed a punctate perinuclear distribution. Moreover, peptide/siRNA complexes achieved high transfection efficiency in vitro without inducing substantial cytotoxicity. We have validated the therapeutic potential of this strategy for cancer treatment by targeting Bcl-2 gene in mouse tumor models, and demonstrated that tumor growth was inhibited. In order to address possible immune side effects of these peptide carriers, biocompatibility study in terms of complement activation and cytokine activation assay were carried out, whereas none of the peptides induced such effects. In conclusion, these results support the potential of these peptides as therapeutic siRNA carrier.
Department of Chemical Engineering and Waterloo Institute for Nanotechnology, University of Waterloo , 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada.
Wen
W
Xu
Ran
R
Pan
Danyang
D
Zhao
Dafeng
D
Chu
Yan
Y
Wu
Rong
R
Wang
Baoling
B
Chen
Yong
Y
Ding
Parisa
P
Sadatmousavi
Yongfang
Y
Yuan
P
P
Chen
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D016466:CHO Cells; D045744:Cell Line, Tumor; D002469:Cell Separation; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D004337:Drug Carriers / Q000737:chemistry*; D015195:Drug Design; D004354:Drug Screening Assays, Antitumor; D011992:Endosomes / Q000737:chemistry*; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D006801:Humans; D008297:Male; D051379:Mice; D018625:Microscopy, Atomic Force; D009368:Neoplasm Transplantation; D010455:Peptides / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*
D001672:Biocompatible Materials; D004337:Drug Carriers; D010455:Peptides; D034741:RNA, Small Interfering
biocompatible; peptide-based carrier; siRNA delivery; tumor inhibition