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Effective Gene Delivery into Human Stem Cells with a Cell-Targeting Peptide-Modified Bioreducible Polymer

 Jagadish Beloor ; Suresh Ramakrishna ; Priti Kumar ; Sang-Kyung Lee ; Sung Wan Kim ; Hyongbum Kim ; HeungSoo Shin ; Hyong Jin Cho ; Sung Hwa Kim ; Jongkil Kim ; Chang Seon Choi ; Kihoon Nam 
 Small, Vol.11(17) : 2069~2079, 2015 
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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.
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1. 연구논문 > 1. College of Medicine > Dept. of Pharmacology
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