title: The role of doxorubicin in non-viral gene transfer in the lung.
abstract: Proteasome inhibitors have been shown to increase adeno-associated virus (AAV)-mediated transduction in vitro and in vivo. To assess if proteasome inhibitors also increase lipid-mediated gene transfer with relevance to cystic fibrosis (CF), we first assessed the effects of doxorubicin and N-acetyl-l-leucinyl-l-leucinal-l-norleucinal in non-CF (A549) and CF (CFTE29o-) airway epithelial cell lines. CFTE29o- cells did not show a response to Dox or LLnL; however, gene transfer in A549 cells increased in a dose-related fashion (p < 0.05), up to approximately 20-fold respectively at the optimal dose (no treatment: 9.3 x 10(4) +/- 1.5 x 10(3), Dox: 1.6 x 10(6)+/-2.6 x 10(5), LLnL: 1.9 x 10(6) +/- 3.2 x 10(5)RLU/mg protein). As Dox is used clinically in cancer chemotherapy we next assessed the effect of this drug on non-viral lung gene transfer in vivo. CF knockout mice were injected intraperitoneally (IP) with Dox (25-100 mg/kg) immediately before nebulisation with plasmid DNA carrying a luciferase reporter gene under the control of a CMV promoter/enhancer (pCIKLux) complexed to the cationic lipid GL67A. Dox also significantly (p < 0.05) increased expression of a plasmid regulated by an elongation factor 1alpha promoter (hCEFI) approximately 8-fold. Although administration of Dox before lung gene transfer may not be a clinically viable option, understanding how Dox increases lung gene expression may help to shed light on intracellular bottle-necks to gene transfer, and may help to identify other adjuncts that may be more appropriate for use in man.
authors.affiliation: Department of Gene Therapy, Imperial College at the National Heart and Lung Institute, Manresa Road, London SW36LR, UK. u.griesenbach@imperial.ac.uk
authors.forename: Uta
authors.initials: U
authors.lastname: Griesenbach
authors.forename: Cuixiang
authors.initials: C
authors.lastname: Meng
authors.forename: Raymond
authors.initials: R
authors.lastname: Farley
authors.forename: Aaron
authors.initials: A
authors.lastname: Gardner
authors.forename: Maresa A
authors.initials: MA
authors.lastname: Brake
authors.forename: Gad M
authors.initials: GM
authors.lastname: Frankel
authors.forename: Dieter C
authors.initials: DC
authors.lastname: Gruenert
authors.forename: Seng H
authors.initials: SH
authors.lastname: Cheng
authors.forename: Ronald K
authors.initials: RK
authors.lastname: Scheule
authors.forename: Eric W F W
authors.initials: EW
authors.lastname: Alton
mesh_terms: D000818:Animals; D002460:Cell Line; D003550:Cystic Fibrosis / Q000235:genetics* / Q000628:therapy*; D004317:Doxorubicin / Q000494:pharmacology*; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000737:chemistry; D006801:Humans; D008168:Lung / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics
chemical_list: D020648:Peptide Elongation Factor 1; D004317:Doxorubicin