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.

Nanoparticle-enhanced generation of gene-transfected mesenchymal stem cells for in vivo cardiac repair

“…BMMSCs are reluctant to accept gene transfection via using nonviral vectors without impacting the pluripotency or cell viability due to the variability in the cell subpopulation and the toxicity of many transfection methods, which frequently results in substantially low efficiencies [15][16][17][18]. Thus, clinical studies have been mostly limited to the non-transfected BMMSCs.…”

Hyaluronic acid reagent functional chitosan-PEI conjugate with AQP2-siRNA suppressed endometriotic lesion formation

“…This result was consistent with the combinations between the ligand and gene vector could enhance gene transfection of the gene delivery system. 37 This therapeutic effect suggested that siRNA might be carried into the cells in cysts and afterward accumulated to the lesion via the encapsulation of (CSO-PEI)HA. It is the (CSO-PEI)HA that facilitate the siRNA to deliver into the target sites.…”

Non-viral gene delivery systems for tissue repair and regeneration

“…The ATS-9R/shFABP4 oligopeptide complex targeted mature adipocytes via binding to inhibitin and silencing of the targeted sequence peptide resulted in reduced lipidosis [ 82 ]. A 29-amino acid cell-binding peptide, RGV, conjugated to the redox-sensitive biodegradable dendrimer PAM-ABP, provided a low toxicity compound that increased transfection rates of both hMSC and hESC (about 60 and 50%, respectively) and retained expression of pluripotent stem cell markers [ 83 ]. Mannosylated CPP was bound to PEI to form the polymer Man-PEI1800-CPP, which was targeted at the mannose receptor on antigen-presenting cells (APC); its transfection rate was higher than 25KDa PEI but with lower toxicity, and in vivo experiments showed that this kind of nVGDS was mainly distributed in the epidermis and dermis [ 52 ].…”