Construction of a tumor cell‑targeting non‑viral gene delivery vector with polyethylenimine modified with RGD sequence‑containing peptide

Xing,Hai‑Bo; Pan,Hong‑Ming; Fang,Yong; Zhou,Xiao‑Yun; Pan,Qin; Li,Da

doi:10.3892/ol.2013.1717

Construction of a tumor cell‑targeting non‑viral gene delivery vector with polyethylenimine modified with RGD sequence‑containing peptide

Authors:
- Hai‑Bo Xing
- Hong‑Ming Pan
- Yong Fang
- Xiao‑Yun Zhou
- Qin Pan
- Da Li
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Affiliations: Department of Intensive Care Unit, Xiasha Hospital, Hangzhou, Zhejiang 310019, P.R. China, Department of Medical Oncology, Sir Run Run Shaw Hospital Affiliated to School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, P.R. China
Published online on: November 29, 2013 https://doi.org/10.3892/ol.2013.1717
Pages: 487-492

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Abstract

The objective of the present study was to construct a novel type of non‑viral gene delivery vector with high delivery efficiency and specific tumor cell‑targeting ability. The CP9 peptide (CYGGRGDTP) containing Arg‑Gly‑Asp sequence was employed to be conjugated onto polyethylenimine (PEI) to act as the role of the targeting moiety. The chemical linker, N‑succinimidyl‑3‑(2‑pyridyldithio) propionate, was applied during the synthesis of the vector (CP9‑PEI). The physicochemical characteristics of the vector were evaluated by the methods of 1H‑nuclear magnetic resonance, Fourier transform infrared spectroscopy, gel retardation assay, electron microscope observation and particle size detection. HepG2 cells were used to verify the gene delivery efficiency and targeting ability by gene delivery procedure and free CP9 peptide inhibition tests. The results showed that the successful synthesis of CP9‑PEI and the synthesized vector may efficiently condense plasmid DNA into round particles with diameters of ~200 nm at a polymer/pDNA ratio of 10. CP9‑PEI may deliver the reporter gene into HepG2 cells with higher efficiency and the efficiency may be inhibited by the free CP9 peptide. The present study suggested that the modification of PEI with the CP9 peptide is an effective method to construct a novel tumor cell‑targeting non‑viral vector, and that the novel vector exhibits great prospect in the field of cancer gene therapy.

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1	Pack DW, Hoffman AS, Pun S and Stayton PS: Design and development of polymers for gene delivery. Nat Rev Drug Discov. 4:581–593. 2005. View Article : Google Scholar : PubMed/NCBI
2	Park J, Singha K, Son S, et al: A review of RGD-functionalized nonviral gene delivery vectors for cancer therapy. Cancer Gene Ther. 19:741–748. 2012. View Article : Google Scholar : PubMed/NCBI
3	Guo XD, Wiradharma N, Liu SQ, et al: Oligomerized alpha-helical KALA peptides with pendant arms bearing cell-adhesion, DNA-binding and endosome-buffering domains as efficient gene transfection vectors. Biomaterials. 33:6284–6291. 2012. View Article : Google Scholar : PubMed/NCBI
4	Katayama K, Furuki R, Yokoyama H, et al: Enhanced in vivo gene transfer into the placenta using RGD fiber-mutant adenovirus vector. Biomaterials. 32:4185–4193. 2011. View Article : Google Scholar : PubMed/NCBI
5	Matsui H, Sakurai F, Katayama K, et al: Enhanced transduction efficiency of fiber-substituted adenovirus vectors by the incorporation of RGD peptides in two distinct regions of the adenovirus serotype 35 fiber knob. Virus Res. 155:48–54. 2011. View Article : Google Scholar
6	Nie Y, Schaffert D, Rödl W, Ogris M, Wagner E and Günther M: Dual-targeted polyplexes: one step towards a synthetic virus for cancer gene therapy. J Control Release. 152:127–134. 2011. View Article : Google Scholar : PubMed/NCBI
7	O’Neill AM, Smith AN, Spangler EA, et al: Resistance of canine lymphoma cells to adenoviral infection due to reduced cell surface RGD binding integrins. Cancer Biol Ther. 11:651–658. 2011.PubMed/NCBI
8	Ginn SL, Alexander IE, Edelstein ML, Abedi MR and Wixon J: Gene therapy clinical trials worldwide to 2012 - an update. J Gene Med. 15:65–77. 2013. View Article : Google Scholar : PubMed/NCBI
9	Boussif O, Lezoualc’h F, Zanta MA, et al: A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci. 92:7297–7301. 1995. View Article : Google Scholar : PubMed/NCBI
10	Huntosova V, Buzova D, Petrovajova D, et al: Development of a new LDL-based transport system for hydrophobic/amphiphilic drug delivery to cancer cells. Int J Pharm. 436:463–471. 2012. View Article : Google Scholar : PubMed/NCBI
11	Doane TL and Burda C: The unique role of nanoparticles in nanomedicine: imaging, drug delivery and therapy. Chem Soc Rev. 41:2885–2911. 2012. View Article : Google Scholar : PubMed/NCBI
12	Zheng M and Zhong Z, Zhou L, Meng F, Peng R and Zhong Z: Poly (ethylene oxide) grafted with short polyethylenimine gives DNA polyplexes with superior colloidal stability, low cytotoxicity, and potent in vitro gene transfection under serum conditions. Biomacromolecules. 13:881–888. 2012. View Article : Google Scholar
13	Nimesh S: Polyethylenimine as a promising vector for targeted iRNA delivery. Curr Clin Pharmacol. 7:121–130. 2012. View Article : Google Scholar
14	Patnaik S and Gupta KC: Novel polyethylenimine-derived nanoparticles for in vivo gene delivery. Expert Opin Drug Deliv. 10:215–228. 2013. View Article : Google Scholar : PubMed/NCBI
15	Jain K, Kesharwani P, Gupta U and Jain NK: A review of glycosylated carriers for drug delivery. Biomaterials. 33:4166–4186. 2012. View Article : Google Scholar : PubMed/NCBI
16	Jiang Q, Shi P, Li C, et al: (Coixan polysaccharide)-graft-polyethylenimine folate for tumor-targeted gene delivery. Macromol Biosci. 11:435–444. 2011. View Article : Google Scholar : PubMed/NCBI
17	Shin JY, Suh D, Kim JM, et al: Low molecular weight polyethylenimine for efficient transfection of human hematopoietic and umbilical cord blood-derived CD34+ cells. Biochim Biophys Acta. 1725:377–384. 2005.PubMed/NCBI
18	Hu Q, Wang J, Shen J, et al: Intracellular pathways and nuclear localization signal peptide-mediated gene transfection by cationic polymeric nanovectors. Biomaterials. 33:1135–1145. 2012. View Article : Google Scholar : PubMed/NCBI
19	Sawant RR, Sriraman SK, Navarro G, Biswas S, Dalvi RA and Torchilin VP: Polyethyleneimine-lipid conjugate-based pH-sensitive micellar carrier for gene delivery. Biomaterials. 33:3942–3951. 2012. View Article : Google Scholar : PubMed/NCBI
20	Munier S, Messai I, Delair T, Verrier B and Ataman-Onal Y: Cationic PLA nanoparticles for DNA delivery: Comparison of three surface polycations for DNA binding, protection and transfection properties. Colloids Surf B Biointerfaces. 43:163–173. 2005. View Article : Google Scholar
21	Stoneham CA, Hollinshead M and Hajitou A: Clathrin-mediated endocytosis and subsequent endo-lysosomal trafficking of adeno-associated virus/phage. J Biol Chem. 287:35849–35859. 2012. View Article : Google Scholar : PubMed/NCBI
22	Kagaya H, Oba M, Miura Y, et al: Impact of polyplex micelles installed with cyclic RGD peptide as ligand on gene delivery to vascular lesions. Gene Ther. 19:61–69. 2012. View Article : Google Scholar : PubMed/NCBI
23	Kim J, Nam HY, Kim TI, et al: Active targeting of RGD-conjugated bioreducible polymer for delivery of oncolytic adenovirus expressing shRNA against IL-8 mRNA. Biomaterials. 32:5158–5166. 2012. View Article : Google Scholar : PubMed/NCBI
24	Wu ZW, Chien CT, Liu CY, Yan JY and Lin SY: Recent progress in copolymer-mediated siRNA delivery. J Drug Target. 20:551–560. 2012. View Article : Google Scholar : PubMed/NCBI