Synthesis and characterization of four-arm poly(ethylene glycol)-based gene delivery vehicles coupled to integrin and DNA-binding peptides

Nicole M. Moore, Tiffany R. Barbour, Shelly E. Sakiyama-Elbert*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

The goal of this study was to develop a gene delivery vehicle that can specifically target cell surface receptors with low nonspecific protein adsorption and low cytotoxicity. Toward this goal, four-arm poly(ethylene glycol) vehicles were functionalized with DNA-binding peptides (DBPs) and integrin-binding (RGD) peptides. We have previously described a novel PEG-based gene delivery vehicle functionalized with DBPs that successfully transfected Chinese hamster ovary (CHO) cells with low toxicity and low protein adsorption. This work investigated whether incorporating RGD peptides onto PEG-DBP vehicles could target specific cell surface receptors and increase transfection efficiency of HEPG2 cells. DBP and RGD peptides were coupled onto PEG-tetraacrylate (PEG-TA) in three combinations (molar ratios of DBP:RGD of 1:3, 2:2, and 3:1) and characterized by measuring particle size, ζ potential, and transfection efficiency as a function of charge ratio (peptide amine groups:DNA phosphate). Nonspecific protein adsorption and cytotoxicity of PEG-DBP-RGD vehicles were also measured. Dynamic light scattering showed that PEG-DBP-RGD vehicles condensed DNA into particles having mean diameters of 250-300 nm and ζ potentials ranging from -10 to 7 mV. It was found that coupling two RGD peptides to the PEG-DBP2 vehicle increased the transfection efficiency at a polymer/DNA charge ratio of 5:1 (+/-) and 6:1 (+/-) and that these vehicles had transfection efficiencies similar to those of polyethylenimine (PEI)/DNA particles. However, coupling one or three RGD peptides to PEG-DBP vehicles did not increase the transfection efficiency. Additionally, the PEG-DBP-RGD/DNA particles adsorbed less protein than PEI particles and were less toxic to HEPG2 cells.

Original languageEnglish (US)
Pages (from-to)140-150
Number of pages11
JournalMolecular Pharmaceutics
Volume5
Issue number1
DOIs
StatePublished - Jan 1 2008

Keywords

  • Gene delivery
  • Integrin
  • Poly(ethylene glycol) vehicles

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmaceutical Science
  • Drug Discovery

Fingerprint Dive into the research topics of 'Synthesis and characterization of four-arm poly(ethylene glycol)-based gene delivery vehicles coupled to integrin and DNA-binding peptides'. Together they form a unique fingerprint.

Cite this