The role of doxorubicin in non-viral gene transfer in the lung |
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| Authors: | Uta Griesenbach Cuixiang Meng Raymond Farley Aaron Gardner Maresa A. Brake Gad M. Frankel Dieter C. Gruenert Seng H. Cheng Ronald K. Scheule Eric W.F.W. Alton |
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| Affiliation: | 1. Department of Gene Therapy, Imperial College at the National Heart and Lung Institute, Manresa Road, London SW3 6LR, UK;2. UK Cystic Fibrosis Gene Therapy Consortium, London, UK;3. Division of Cell and Molecular Biology, Imperial College London, London SW7 2AZ, UK;4. California Pacific Medical Center Research Institute, San Francisco, CA 94107, USA;5. Department of Laboratory Medicine, University of California, San Francisco, San Francisco, CA 94143, USA;6. Department of Medicine, University of Vermont School of Medicine, Burlington, VT 05405, USA;7. Genzyme Ltd, Haverhill, UK;1. Laboratory of Cellular Biology, Department of Biology, ICB, Federal University of Juiz de Fora, UFJF, Rua José Lourenço Kelmer, Juiz de Fora, MG 36036-900, Brazil;2. Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, CLS 943, Boston, MA 02215, USA;1. Translational Research Unit, Research and Development Division, Kyowa Hakko Kirin, Co., Ltd., Japan;2. Research Functions Unit, Research and Development Division, Kyowa Hakko Kirin, Co., Ltd., Japan;3. Veterinary Pathology, Osaka Prefecture University, Japan;1. Department of Pediatrics, Division of Cellular and Molecular Therapy, University of Florida, Gainesville, FL;2. Center of Molecular and Cellular Therapeutics, The Children''s Hospital of Philadelphia, Philadelphia, PA;3. Department of Pathology, Immunology and Laboratory Medicine, University of Florida, Gainesville, FL;4. Division of Immunology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA;1. Departments of Respiratory Medicine, The First Affiliated Hospital of Dalian Medical University, 222# Zhongshan Road, Dalian, Liaoning 116011, PR China;2. Departments of Nephrology, The First Affiliated Hospital of Dalian Medical University, 222# Zhongshan Road, Dalian, Liaoning 116011, PR China;1. Department of Cardiology, Shanghai East hospital, Tongji University, Shanghai, China;2. Department of Cardiac Surgery, Shanghai East hospital, Tongji University, Shanghai, China;1. Department Pediatrics, University of Florida, Gainesville, FL, USA;2. Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN, USA;3. Department of Pathology, Immunology and Laboratory Medicine, University of Florida, Gainesville, FL, USA;4. Division of Immunology, Beth Israel Deaconess Medical Center (BIDMC), Harvard Medical School, Boston, MA, USA |
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| 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. |
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