Dilution-stable PAMAM G1-grafted polyrotaxane supermolecules deliver gene into cells through a caveolae-dependent pathway.

Abstract

Numerous preclinical studies have demonstrated that polycation mediated gene delivery systems successfully achieved efficient gene transfer into cells and animal models. However, results of their clinical trials to date have been disappointing. That self-assembled gene and polycation systems should be stable undergoing dilution in the body is one of the prerequisites to ensuring efficiency of gene transfer in clinical trials, but it was neglected in most preclinical studies. In this account, we developed the dilution-stable PAMAM G1-grafted polyrotaxane (PPG1) supermolecules in which PAMAM G1-grafted α-cyclodextrins are threaded onto a PEG chain capped with hydrophobic adamantanamine. The PPG1/pDNA polyplex (approximate 100 nm in diameter) was very stable and kept its initial particle size and a uniform size distribution at ultrahigh dilution, whereas DNA/PEI 25K polyplex was above three times bigger at a 16-fold dilution than the initial size and their particle size distribution indicated multiple peaks mainly due to forming loose and noncompacted aggregates. PPG1 supermolecules showed significantly superior transfection efficiencies compared to either PEI 25K or Lipofectamine 2000 in most cell lines tested including normal cells (HEK293A) and cancer cells (Bel7402, HepG2, and HeLa). Furthermore, we found that the PPG1 supermolecules delivered DNA into HEK293A through a caveolae-dependent pathway but not a clathrin-dependent pathway as PEI 25K did. These findings raised the intriguing possibility that the caveolae-dependent pathway of PPG1 supermolecule/pDNA polyplex avoiding lysosomal degradation was attributed to their high transfection efficiency. The dilution-stable PPG1 supermolecule polyplex facilitating caveolae-dependent internalization has potential applications to surmount the challenges of high dilutions in the body and lysosomal degradation faced by most gene therapy clinical trials.

• Authors: Huan Huang, Duanwen Cao, Linghao Qin, Shouqin Tian, Yang Liang, Shirong Pan, Min Feng
• Journal: Molecular pharmaceutics (Mol Pharm) Vol. 11 Issue 7 Pg. 2323-33 (Jul 07 2014) ISSN: 1543-8392 [Electronic] United States
• PMID: 24957192 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Clathrin
• Cyclodextrins
• Dendrimers
• PAMAM Starburst
• Polyamines
• Polyelectrolytes
• Rotaxanes
• alpha-Cyclodextrins
• polycations
• polyrotaxane
• Poloxamer
• Polyethylene Glycols
• DNA

Topics

• Caveolae (chemistry)
• Cell Line
• Cell Line, Tumor
• Clathrin (chemistry)
• Cyclodextrins (chemistry)
• DNA (chemistry)
• Dendrimers (chemistry)
• Gene Transfer Techniques
• HEK293 Cells
• HeLa Cells
• Hep G2 Cells
• Humans
• Particle Size
• Poloxamer (chemistry)
• Polyamines (chemistry)
• Polyelectrolytes
• Polyethylene Glycols (chemistry)
• Rotaxanes (chemistry)
• Transfection (methods)
• alpha-Cyclodextrins (chemistry)