Design of a multicomponent peptide-woven nanocomplex for delivery of siRNA

Eunsung Jun; Soyoun Kim; Jong-Ho Kim; Kiweon Cha; In-Seop So; Hye-Nam Son; Byung-Heon Lee; Kwangmeyung Kim; Ick Chan Kwon; Sang Yoon Kim; In-San Kim

doi:10.1371/journal.pone.0118310

Design of a multicomponent peptide-woven nanocomplex for delivery of siRNA

PLoS One. 2015 Feb 23;10(2):e0118310. doi: 10.1371/journal.pone.0118310. eCollection 2015.

Authors

Eunsung Jun¹, Soyoun Kim², Jong-Ho Kim³, Kiweon Cha⁴, In-Seop So², Hye-Nam Son², Byung-Heon Lee², Kwangmeyung Kim⁵, Ick Chan Kwon⁵, Sang Yoon Kim⁶, In-San Kim⁵

Affiliations

¹ Department of Biochemistry and Cell Biology, Cell and Matrix Research Institute, School of Medicine, Kyungpook National University, Daegu, Republic of Korea; Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul, Republic of Korea.
² Department of Biochemistry and Cell Biology, Cell and Matrix Research Institute, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
³ Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University, Seoul, Republic of Korea.
⁴ Division of high-risk pathogen research, Korea National Institute of Health, Korea Centers For Disease Control & Prevention (KCDC), Osong, Chungbuk, Republic of Korea.
⁵ Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul, Republic of Korea.
⁶ Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul, Republic of Korea; Department of Otolaryngology, University of Ulsan College of Medicine, Seoul, Republic of Korea.

Abstract

We developed and tested a multicomponent peptide-woven siRNA nanocomplex (PwSN) comprising different peptides designed for efficient cellular targeting, endosomal escape, and release of siRNA. To enhance tumor-specific cellular uptake, we connected an interleukin-4 receptor-targeting peptide (I4R) to a nine-arginine peptide (9r), yielding I4R-9r. To facilitate endosomal escape, we blended endosomolytic peptides into the I4R-9r to form a multicomponent nanocomplex. Lastly, we modified 9r peptides by varying the number and positions of positive charges to obtain efficient release of siRNA from the nanocomplex in the cytosol. Using this step-wise approach for overcoming the biological challenges of siRNA delivery, we obtained an optimized PwSN with significant biological activity in vitro and in vivo. Interestingly, surface plasmon resonance analyses and three-dimensional peptide models demonstrated that our designed peptide adopted a unique structure that was correlated with faster complex disassembly and a better gene-silencing effect. These studies further elucidate the siRNA nanocomplex delivery pathway and demonstrate the applicability of our stepwise strategy to the design of siRNA carriers capable of overcoming multiple challenges and achieving efficient delivery.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenocarcinoma / genetics
Adenocarcinoma / metabolism
Adenocarcinoma / pathology
Amino Acid Sequence
Animals
Cell Survival / drug effects
Colorectal Neoplasms / genetics
Colorectal Neoplasms / metabolism
Colorectal Neoplasms / pathology
Drug Delivery Systems / methods*
HT29 Cells
HeLa Cells
Humans
Luciferases / genetics
Luciferases / metabolism
Male
Mice, Inbred BALB C
Mice, Nude
Molecular Sequence Data
Nanoparticles / administration & dosage
Nanoparticles / chemistry*
Peptides / chemistry*
Peptides / genetics
RNA, Small Interfering / administration & dosage*
RNA, Small Interfering / chemistry
RNA, Small Interfering / genetics
Receptors, Interleukin-4 / chemistry
Receptors, Interleukin-4 / genetics
Reproducibility of Results
Surface Plasmon Resonance
Transplantation, Heterologous

Substances

Peptides
RNA, Small Interfering
Receptors, Interleukin-4
Luciferases

Grants and funding

This work was supported by the National Research Foundation of Korea(NRF) grant funded by the Korea government(MEST) (2010-0029206); by the National Research Foundation of Korea(NRF) grant funded by the Korea government (2014R1A5A2009242); by the KIST Institutional Program (Project No. 2E25000). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.