Inefficient cationic lipid-mediated siRNA and antisense oligonucleotide transfer to airway epithelial cells in vivo

详细信息    查看全文

• 作者：Uta Griesenbach (1) (10)
  Chris Kitson (2)
  Escudero Sara Garcia (1) (10)
  Raymond Farley (1) (10)
  Charanjit Singh (1) (10)
  Luci Somerton (1) (10)
  Hazel Painter (3)
  Rbecca L Smith (3)
  Deborah R Gill (3)
  Stephen C Hyde (3)
  Yu-Hua Chow (4)
  Jim Hu (4)
  Mike Gray (5)
  Mark Edbrooke (2)
  Varrie Ogilvie (6)
  Gordon MacGregor (6)
  Ronald K Scheule (7)
  Seng H Cheng (7)
  Natasha J Caplen (8) (9)
  Eric WFW Alton (1) (10)
  
• 刊名：Respiratory Research
• 出版年：2006
• 出版时间：December 2006
• 年：2006
• 卷：7
• 期：1
• 全文大小：1658KB
• 参考文献：1. [http://www.isispharm.com]
  2. [http://www.isispharm.com]
  3. McManaway ME, Neckers LM, Loke SL, al Nasser AA, Redner RL, Shiramizu BT, Goldschmidts WL, Huber BE, Bhatia K, Magrath IT: Tumour-specific inhibition of lymphoma growth by an antisense oligodeoxynucleotide. / Lancet 1990, 335:808-11. CrossRef
  4. Kulka M, Smith CC, Aurelian L, Fishelevich R, Meade K, Miller P, Ts'o PO: Site specificity of the inhibitory effects of oligo(nucleoside methylphosphonate)s complementary to the acceptor splice junction of herpes simplex virus type 1 immediate early mRNA 4. / Proc Natl Acad Sci USA 1989, 86:6868-872. CrossRef
  5. Caplen NJ, Parrish S, Imani F, Fire A, Morgan RA: Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. / Proc Natl Acad Sci USA 2001, 98:9742-747. CrossRef
  6. Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T: Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. / Nature 2001, 411:494-98. CrossRef
  7. Schildow DV, Friel SB: Cystic Fibrosis. In / Clinical Respiratory Medicine. Mosby USA; 2004:19-9.
  8. Rochelle LG, Li DC, Ye H, Lee E, Talbot CR, Boucher RC: Distribution of ion transport mRNAs throughout murine nose and lung. / Am J Physiol Lung Cell Mol Physiol 2000, 279:L14-L24.
  9. Reddy MM, Light MJ, Quinton PM: Activation of the epithelial Na+ channel (ENaC) requires CFTR Cl- channel function. / Nature 1999, 402:301-04. CrossRef
  10. Konig J, Schreiber R, Voelcker T, Mall M, Kunzelmann K: The cystic fibrosis transmembrane conductance regulator (CFTR) inhibits ENaC through an increase in the intracellular Cl- concentration. / EMBO Rep 2001, 2:1047-051. kve232">CrossRef
  11. Hofmann T, Stutts MJ, Ziersch A, Ruckes C, Weber WM, Knowles MR, Lindemann H, Boucher RC: Effects of topically delivered benzamil and amiloride on nasal potential difference in cystic fibrosis. / Am J Respir Crit Care Med 1998, 157:1844-849.
  12. Jain L, Chen XJ, Malik B, Al Khalili O, Eaton DC: Antisense oligonucleotides against the alpha-subunit of ENaC decrease lung epithelial cation-channel activity. / Am J Physiol 1999, 276:L1046-L1051.
  13. Ferrari S, Geddes DM, Alton EW: Barriers to and new approaches for gene therapy and gene delivery in cystic fibrosis. / Adv Drug Deliv Rev 2002, 54:1373-393. CrossRef
  14. Crooke ST, Lemonidis KM, Neilson L, Griffey R, Lesnik EA, Monia BP: Kinetic characteristics of Escherichia coli RNase H1: cleavage of various antisense oligonucleotide-RNA duplexes. / Biochem J 1995, 312:599-08.
  15. Lee ER, Marshall J, Siegel CS, Jiang C, Yew NS, Nichols MR, Nietupski JB, Ziegler RJ, Lane MB, Wang KX, Wan NC, Scheule RK, Harris DJ, Smith AE, Cheng SH: Detailed analysis of structures and formulations of cationic lipids for efficient gene transfer to the lung. / Hum Gene Ther 1996, 7:1701-717. CrossRef
  16. Smith SN, Middleton PG, Chadwick S, Jaffe A, Bush KA, Rolleston S, Farley R, Delaney SJ, Wainwright B, Geddes DM, Alton EW: The in vivo effects of milrinone on the airways of cystic fibrosis mice and human subjects. / Am J Respir Cell Mol Biol 1999, 20:129-34.
  17. Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. / Methods 2001, 25:402-08. CrossRef
  18. Eastman SJ, Lukason MJ, Tousignant JD, Murray H, Lane MD, St George JA, Akita GY, Cherry M, Cheng SH, Scheule RK: A concentrated and stable aerosol formulation of cationic lipid:DNA complexes giving high-level gene expression in mouse lung. / Hum Gene Ther 1997, 8:765-73. CrossRef
  19. Howard DP, Cuffe JE, Boyd CA, Korbmacher C: L-arginine effects on Na+ transport in M-1 mouse cortical collecting duct cells -a cationic amino acid absorbing epithelium. / J Membr Biol 2001, 180:111-21. CrossRef
  20. Alton EW, Stern M, Farley R, Jaffe A, Chadwick SL, Phillips J, Davies J, Smith SN, Browning J, Davies MG, Hodson ME, Durham SR, Li D, Jeffery PK, Scallan M, Balfour R, Eastman SJ, Cheng SH, Smith AE, Meeker D, Geddes DM: Cationic lipid-mediated CFTR gene transfer to the lungs and nose of patients with cystic fibrosis: a double-blind placebo-controlled trial. / Lancet 1999, 353:947-54. CrossRef
  21. Ruiz FE, Clancy JP, Perricone MA, Bebok Z, Hong JS, Cheng SH, Meeker DP, Young KR, Schoumacher RA, Weatherly MR, Wing L, Morris JE, Sindel L, Rosenberg M, van Ginkel FW, McGhee JR, Kelly D, Lyrene RK, Sorscher EJ: A clinical inflammatory syndrome attributable to aerosolized lipid-DNA administration in cystic fibrosis. / Hum Gene Ther 2001, 12:751-61. CrossRef
  22. Chow YH, O'Brodovich H, Plumb J, Wen Y, Sohn KJ, Lu Z, Zhang F, Lukacs GL, Tanswell AK, Hui CC, Buchwald M, Hu J: Development of an epithelium-specific expression cassette with human DNA regulatory elements for transgene expression in lung airways. / Proc Natl Acad Sci USA 1997, 94:14695-4700. CrossRef
  23. Vickers TA, Koo S, Bennett CF, Crooke ST, Dean NM, Baker BF: Efficient reduction of target RNAs by small interfering RNA and RNase H-dependent antisense agents. A comparative analysis. / J Biol Chem 2003, 278:7108-118. CrossRef
  24. Zhang X, Shan P, Jiang D, Noble PW, Abraham NG, Kappas A, Lee PJ: Small interfering RNA targeting heme oxygenase-1 enhances ischemia-reperfusion-induced lung apoptosis. / J Biol Chem 2004, 279:10677-0684. CrossRef
  25. Ge Q, McManus MT, Nguyen T, Shen CH, Sharp PA, Eisen HN, Chen J: RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription. / Proc Natl Acad Sci USA 2003, 100:2718-723. CrossRef
  26. Tompkins SM, Lo CY, Tumpey TM, Epstein SL: Protection against lethal influenza virus challenge by RNA interference in vivo. / Proc Natl Acad Sci USA 2004, 101:8682-686. CrossRef
  27. Zhang W, Yang H, Kong X, Mohapatra S, Juan-Vergara H, Hellermann G, Behera S, Singam R, Lockey RF, Mohapatra SS: Inhibition of respiratory syncytial virus infection with intranasal siRNA nanoparticles targeting the viral NS1 gene. / Nat Med 2005, 11:56-2. CrossRef
  28. Staub O, Gautschi I, Ishikawa T, Breitschopf K, Ciechanover A, Schild L, Rotin D: Regulation of stability and function of the epithelial Na+ channel (ENaC) by ubiquitination. / EMBO J 1997, 16:6325-36. CrossRef
  29. Valentijn JA, Fyfe GK, Canessa CM: Biosynthesis and processing of epithelial sodium channels in Xenopus oocytes. / J Biol Chem 1998, 273:30344-0351. CrossRef
  30. Templin MV, Levin AA, Graham MJ, Aberg PM, Axelsson BI, Butler M, Geary RS, Bennett CF: Pharmacokinetic and toxicity profile of a phosphorothioate oligonucleotide following inhalation delivery to lung in mice. / Antisense Nucleic Acid Drug Dev 2000, 10:359-68.
  31. Singh PK, Tack BF, McCray PB Jr, Welsh MJ: Synergistic and additive killing by antimicrobial factors found in human airway surface liquid. / Am J Physiol Lung Cell Mol Physiol 2000, 279:L799-L805.
  32. Tate S, MacGregor G, Davis M, Innes JA, Greening AP: Airways in cystic fibrosis are acidified: detection by exhaled breath condensate. / Thorax 2002, 57:926-29. CrossRef
  33. Effros RM, Biller J, Foss B, Hoagland K, Dunning MB, Castillo D, Bosbous M, Sun F, Shaker R: A simple method for estimating respiratory solute dilution in exhaled breath condensates. / Am J Respir Crit Care Med 2003, 168:1500-505. CrossRef
  34. Eastman SJ, Lukason MJ, Tousignant JD, Murray H, Lane MD, St George JA, Akita GY, Cherry M, Cheng SH, Scheule RK: A concentrated and stable aerosol formulation of cationic lipid:DNA complexes giving high-level gene expression in mouse lung. / Hum Gene Ther 1997, 8:765-73. CrossRef
  
• 作者单位：Uta Griesenbach (1) (10)
  Chris Kitson (2)
  Escudero Sara Garcia (1) (10)
  Raymond Farley (1) (10)
  Charanjit Singh (1) (10)
  Luci Somerton (1) (10)
  Hazel Painter (3)
  Rbecca L Smith (3)
  Deborah R Gill (3)
  Stephen C Hyde (3)
  Yu-Hua Chow (4)
  Jim Hu (4)
  Mike Gray (5)
  Mark Edbrooke (2)
  Varrie Ogilvie (6)
  Gordon MacGregor (6)
  Ronald K Scheule (7)
  Seng H Cheng (7)
  Natasha J Caplen (8) (9)
  Eric WFW Alton (1) (10)
  
  1. Department of Gene Therapy, Faculty of Medicine at the National Heart and Lung Institute, Imperial College, London, UK
  10. UK Cystic Fibrosis Gene Therapy Consortium, Bonner Road, London, E2 9JX, UK
  2. GlaxoSmithKline, eat West Road, Brentford, Middlesex, TW8 9GS, United Kingdom
  3. Gene Medicine Research Group, Nuffield Department of Clinical Laboratory Sciences, John Radcliffe Hospital, University of Oxford, UK
  4. Programme in Lung Biology Research, Hospital for Sick Children and Department of Laboratory Medicine and Pathobiology, University of Toronto, 3359, Mississauga, ON L5L 1C6
  5. Institute for Cell and Molecular Biosciences, University Medical School, Newcastle, UK
  6. Medical Genetics Section, University of Edinburgh, Edinburgh, UK
  7. Genzyme Corporation, USA
  8. Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892
  9. Gene Silencing Section, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892
  

文摘

Background The cationic lipid Genzyme lipid (GL) 67 is the current "gold-standard" for in vivo lung gene transfer. Here, we assessed, if GL67 mediated uptake of siRNAs and asODNs into airway epithelium in vivo. Methods Anti-lacZ and ENaC (epithelial sodium channel) siRNA and asODN were complexed to GL67 and administered to the mouse airway epithelium in vivo Transfection efficiency and efficacy were assessed using real-time RT-PCR as well as through protein expression and functional studies. In parallel in vitro experiments were carried out to select the most efficient oligonucleotides. Results In vitro, GL67 efficiently complexed asODNs and siRNAs, and both were stable in exhaled breath condensate. Importantly, during in vitro selection of functional siRNA and asODN we noted that asODNs accumulated rapidly in the nuclei of transfected cells, whereas siRNAs remained in the cytoplasm, a pattern consistent with their presumed site of action. Following in vivo lung transfection siRNAs were only visible in alveolar macrophages, whereas asODN also transfected alveolar epithelial cells, but no significant uptake into conducting airway epithelial cells was seen. SiRNAs and asODNs targeted to β-galactosidase reduced βgal mRNA levels in the airway epithelium of K18-lacZ mice by 30% and 60%, respectively. However, this was insufficient to reduce protein expression. In an attempt to increase transfection efficiency of the airway epithelium, we increased contact time of siRNA and asODN using the in vivo mouse nose model. Although highly variable and inefficient, transfection of airway epithelium with asODN, but not siRNA, was now seen. As asODNs more effectively transfected nasal airway epithelial cells, we assessed the effect of asODN against ENaC, a potential therapeutic target in cystic fibrosis; no decrease in ENaC mRNA levels or function was detected. Conclusion This study suggests that although siRNAs and asODNs can be developed to inhibit gene expression in culture systems and certain organs in vivo, barriers to nucleic acid transfer in airway epithelial cells seen with large DNA molecules may also affect the efficiency of in vivo uptake of small nucleic acid molecules.