title: Effective gene delivery into human stem cells with a cell-targeting Peptide-modified bioreducible polymer.
abstract: Stem cells are poorly permissive to non-viral gene transfection reagents. In this study, we explored the possibility of improving gene delivery into human embryonic (hESC) and mesenchymal (hMSC) stem cells by synergizing the activity of a cell-binding ligand with a polymer that releases nucleic acids in a cytoplasm-responsive manner. A 29 amino acid long peptide, RVG, targeting the nicotinic acetylcholine receptor (nAchR) was identified to bind both hMSC and H9-derived hESC. Conjugating RVG to a redox-sensitive biodegradable dendrimer-type arginine-grafted polymer (PAM-ABP) enabled nanoparticle formation with plasmid DNA without altering the environment-sensitive DNA release property and favorable toxicity profile of the parent polymer. Importantly, RVG-PAM-ABP quantitatively enhanced transfection into both hMSC and hESC compared to commercial transfection reagents like Lipofectamine 2000 and Fugene. ∼60% and 50% of hMSC and hESC were respectively transfected, and at increased levels on a per cell basis, without affecting pluripotency marker expression. RVG-PAM-ABP is thus a novel bioreducible, biocompatible, non-toxic, synthetic gene delivery system for nAchR-expressing stem cells. Our data also demonstrates that a cell-binding ligand like RVG can cooperate with a gene delivery system like PAM-ABP to enable transfection of poorly-permissive cells.
authors.affiliation: Department of Internal Medicine, Section of Infectious Diseases, Yale University School of Medicine, New Haven, CT, 06520, USA; Department of Bioengineering and Institute of Nano Science and Technology, Hanyang University, Seoul, 133-791, Korea.
authors.forename: Jagadish
authors.initials: J
authors.lastname: Beloor
authors.forename: Suresh
authors.initials: S
authors.lastname: Ramakrishna
authors.forename: Kihoon
authors.initials: K
authors.lastname: Nam
authors.forename: Chang
authors.initials: C
authors.lastname: Seon Choi
authors.forename: Jongkil
authors.initials: J
authors.lastname: Kim
authors.forename: Sung Hwa
authors.initials: SH
authors.lastname: Kim
authors.forename: Hyong Jin
authors.initials: HJ
authors.lastname: Cho
authors.forename: HeungSoo
authors.initials: H
authors.lastname: Shin
authors.forename: Hyongbum
authors.initials: H
authors.lastname: Kim
authors.forename: Sung Wan
authors.initials: SW
authors.lastname: Kim
authors.forename: Sang-Kyung
authors.initials: SK
authors.lastname: Lee
authors.forename: Priti
authors.initials: P
authors.lastname: Kumar
mesh_terms: D000818:Animals; D001120:Arginine / Q000737:chemistry; D001672:Biocompatible Materials / Q000737:chemistry*; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006801:Humans; D008024:Ligands; D008055:Lipids / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology; D051379:Mice; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry*; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D011108:Polymers / Q000737:chemistry; D011978:Receptors, Nicotinic / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*; D014162:Transfection
chemical_list: D001672:Biocompatible Materials; C411955:FuGene; D008024:Ligands; D008055:Lipids; C086724:Lipofectamine; D010455:Peptides; D011108:Polymers; D011978:Receptors, Nicotinic; D049452:Green Fluorescent Proteins; D004247:DNA; D001120:Arginine
keywords: bioreducible polymer; cell-binding ligand; embryonic stem cells; mesenchymal stem cells; non-viral gene delivery