title: Gene delivery by dendrimers operates via different pathways in different cells, but is enhanced by the presence of caveolin.
abstract: In order to optimise and improve the efficacy of transfection mediated by dendrimers, it is essential to fully understand the mechanisms of cell entry and intracellular trafficking by these complexes. Previously, we have shown that gene delivery by dendrimers is dependent from cholesterol and membrane rafts. The inhibition of transfection by treatment with filipin III suggested that gene delivery might be occurring by a caveolin-dependent pathway. We therefore investigated the internalisation and transfection properties of dendriplexes using cell lines (HeLa and HepG2) that express few caveolae. We show that, in contrast to other cells, cholesterol depletion does not affect the ability of dendriplexes to transfect these cells. Inhibition of clathrin-independent, phagocytic and macropinocytic pathways also failed to inhibit transfection of these cells and endothelial cells. However, overexpression of caveolin 1 resulted in an increased rate of dendriplex uptake into HeLa, HepG2 and endothelial cells, and increased transfection efficiency. Furthermore, in endothelial cells, confocal microscopy demonstrated colocalisation of dendriplexes and caveolin 1. These data highlight that dendriplexes may use different internalisation pathways in different cells, and that caveolae form a preferential route for gene delivery by this non-viral vector.
authors.affiliation: Department of Immunology, Division of Medicine, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London, W12 0NN, United Kingdom.
authors.forename: Maria
authors.initials: M
authors.lastname: Manunta
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Nichols
authors.forename: Peng Hong
authors.initials: PH
authors.lastname: Tan
authors.forename: Pervinder
authors.initials: P
authors.lastname: Sagoo
authors.forename: Jennifer
authors.initials: J
authors.lastname: Harper
authors.forename: Andrew J T
authors.initials: AJ
authors.lastname: George
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D051242:Caveolin 1 / Q000378:metabolism* / Q000502:physiology; D002460:Cell Line; D045744:Cell Line, Tumor; D002746:Chlorpromazine / Q000494:pharmacology; D002784:Cholesterol / Q000737:chemistry; D002867:Chromones / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D050091:Dendrimers / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D042783:Endothelial Cells; D006367:HeLa Cells; D006801:Humans; D009025:Morpholines / Q000494:pharmacology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D014162:Transfection / Q000379:methods*; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D051242:Caveolin 1; D002867:Chromones; D050091:Dendrimers; D009025:Morpholines; D019384:Nucleic Acid Synthesis Inhibitors; D015638:Cytochalasin D; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D002784:Cholesterol; D002746:Chlorpromazine; D000077191:Wortmannin