Formulation of glutathione responsive anti-proliferative nanoparticles from thiolated Akt1 siRNA and disulfide-crosslinked PEI for efficient anti-cancer gene therapy

Muthunarayanan Muthiah; Hui-Lian Che; Santhosh Kalash; Jihoon Jo; Seok-Yong Choi; Won Jong Kim; Chong Su Cho; Jae Young Lee; In-Kyu Park

doi:10.1016/j.colsurfb.2014.12.022

Formulation of glutathione responsive anti-proliferative nanoparticles from thiolated Akt1 siRNA and disulfide-crosslinked PEI for efficient anti-cancer gene therapy

Colloids Surf B Biointerfaces. 2015 Feb 1:126:322-7. doi: 10.1016/j.colsurfb.2014.12.022. Epub 2014 Dec 31.

Authors

Muthunarayanan Muthiah¹, Hui-Lian Che¹, Santhosh Kalash¹, Jihoon Jo¹, Seok-Yong Choi¹, Won Jong Kim², Chong Su Cho³, Jae Young Lee⁴, In-Kyu Park⁵

Affiliations

¹ Department of Biomedical Sciences, BK21 PLUS Center for Creative Biomedical Scientists at Chonnam National University, Research Institute of Medical Sciences, Chonnam National University Medical School, Gwangju 501-757, South Korea.
² Department of Chemistry, Pohang University of Science and Technology, Pohang 790-784, South Korea.
³ Department of Agricultural Biotechnology, Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921, South Korea.
⁴ School of Materials Science and Engineering, Gwangju Institute of Science and Technology, Gwangju 500-712, South Korea.
⁵ Department of Biomedical Sciences, BK21 PLUS Center for Creative Biomedical Scientists at Chonnam National University, Research Institute of Medical Sciences, Chonnam National University Medical School, Gwangju 501-757, South Korea. Electronic address: [email protected].

PMID: 25576812
DOI: 10.1016/j.colsurfb.2014.12.022

Abstract

In this study, thiol-modified siRNA (SH-siRNA) was delivered by bioreducible polyethylenimine (ssPEI), to enhance physicochemical properties of polyplexes and function of siRNA through disulfide bonding between SH-siRNA and ssPEI. The ssPEI was utilized to deliver Akt1 SH-siRNA for suppression of Akt1 mRNA and blockage of Akt1 protein translation, resulting in reduced cellular proliferation and the induction of apoptosis. Disulfide bondings between the ssPEI and SH-siRNA through thiol groups in both were confirmed by DTT treatment. Complexation between ssPEI and Akt1SH-siRNA was enhanced and reduced surface charge of ssPEI/Akt1SH-siRNA complexes with smaller average particle sizes even at lower N/P ratios was obtained compared with PEI/Akt1siRNA ones. Cellular uptake of ssPEI/Akt1SH-siRNA complexes in CT-26 mouse colon cancer cells was also enhanced. The ssPEI/Akt1SH-siRNA complexes reduced proliferation and increased apoptosis of mouse colon cancer cells in vitro. In an in vivo mouse tumor model, the complexes reduced tumor proliferation and downregulation of Akt1 compared to controls.

Keywords: Akt1; Bioreducible PEI; Colon cancer; Thiol modification; Tumor suppression; siRNA.

Publication types

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

MeSH terms

Animals
Apoptosis / drug effects
Cell Line, Tumor
Cell Proliferation / drug effects
Cell Survival / drug effects
Colonic Neoplasms / genetics
Colonic Neoplasms / pathology
Colonic Neoplasms / therapy
Cross-Linking Reagents / chemistry
Disease Models, Animal
Disulfides / chemistry*
Genetic Therapy*
Glutathione / chemistry*
Mice
Mice, Inbred BALB C
Nanoparticles / chemistry*
Polyethyleneimine / chemical synthesis
Polyethyleneimine / chemistry*
Proto-Oncogene Proteins c-akt / biosynthesis
Proto-Oncogene Proteins c-akt / genetics*
RNA, Small Interfering / chemistry*
RNA, Small Interfering / genetics
RNA, Small Interfering / therapeutic use
Sulfhydryl Compounds / chemistry

Substances

Cross-Linking Reagents
Disulfides
RNA, Small Interfering
Sulfhydryl Compounds
Polyethyleneimine
Proto-Oncogene Proteins c-akt
Glutathione