The role of doxorubicin in non-viral gene transfer in the lung

• Published in: Biomaterials 2009-04, Vol.30 (10), p.1971-1977
• Main Authors: Griesenbach, Uta, Meng, Cuixiang, Farley, Raymond, Gardner, Aaron, Brake, Maresa A, Frankel, Gad M, Gruenert, Dieter C, Cheng, Seng H, Scheule, Ronald K, Alton, Eric W.F.W
• Format: Article
• Language: English
• Subjects: Adeno-associated virus; Advanced Basic Science; Animals; Cell Line; Cystic Fibrosis - genetics; Cystic Fibrosis - therapy; Cytomegalovirus; Dentistry; Doxorubicin - pharmacology; Female; Gene transfer; Gene Transfer Techniques; Genetic Vectors - administration & dosage; Genetic Vectors - chemistry; Humans; In vitro test; In vivo test; Lipid; Lung; Lung - drug effects; Lung - metabolism; Lung - pathology; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Peptide Elongation Factor 1 - genetics; Promoter Regions, Genetic - genetics
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2008.12.037

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 × 104 ± 1.5 × 103 , Dox: 1.6 × 106 ± 2.6 × 105 , LLnL: 1.9 × 106 ± 3.2 × 105 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 1α 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.