Pharmaceutical Research

, Volume 28, Issue 4, pp 694–701 | Cite as

Hydrodynamic Gene Delivery and Its Applications in Pharmaceutical Research

  • Barbara Bonamassa
  • Li Hai
  • Dexi Liu
Expert Review


Hydrodynamic delivery has emerged as the simplest and most effective method for intracellular delivery of membrane-impermeable substances in rodents. The system employs a physical force generated by a rapid injection of large volume of solution into a blood vessel to enhance the permeability of endothelium and the plasma membrane of the parenchyma cells to allow delivery of substance into cells. The procedure was initially established for gene delivery in mice, and its applications have been extended to the delivery of proteins, oligo nucleotides, genomic DNA and RNA sequences, and small molecules. The focus of this review is on applications of hydrodynamic delivery in pharmaceutical research. Examples are provided to highlight the use of hydrodynamic delivery for study of transcriptional regulation of CYP enzymes, for establishment of animal model for viral infections, and for gene drug discovery and gene function analysis.


gene therapy hydrodynamic delivery nonviral gene delivery oligo nucleotides protein drug discovery siRNA 



This work was supported in part by NIH grants RO1EB007357 and RO1HL098295. B. Bonamassa is recipient of the Rotary Foundation Ambassadorial Scholarship for the 2009/2010 academic year.


  1. 1.
    Gao X, Kim KS, Liu D. Nonviral gene delivery: what we know and what is next. AAPS J. 2007;9:E92–104.PubMedCrossRefGoogle Scholar
  2. 2.
    Herweijer H, Wolff JA. Gene therapy progress and prospects: hydrodynamic gene delivery. Gene Ther. 2007;14:99–107.PubMedGoogle Scholar
  3. 3.
    Kobayashi N, Nishikawa M, Takakura Y. The hydrodynamics-based procedure for controlling the pharmacokinetics of gene medicines at whole body, organ and cellular levels. Adv Drug Deliv Rev. 2005;57:713–31.PubMedCrossRefGoogle Scholar
  4. 4.
    Budker V, Zhang G, Knechtle S, Wolff JA. Naked DNA delivered intraportally expresses efficiently in hepatocytes. Gene Ther. 1996;3:593–8.PubMedGoogle Scholar
  5. 5.
    Zhang G, Vargo D, Budker V, Armstrong N, Knechtle S, Wolff JA. Expression of naked plasmid DNA injected into the afferent and efferent vessels of rodent and dog livers. Hum Gene Ther. 1997;8:1763–72.PubMedCrossRefGoogle Scholar
  6. 6.
    Liu F, Song Y, Liu D. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther. 1999;6:1258–66.PubMedCrossRefGoogle Scholar
  7. 7.
    Zhang G, Budker V, Wolff JA. High levels of foreign gene expression in hepatocytes after tail vein injections of naked plasmid DNA. Hum Gene Ther. 1999;10:1735–7.PubMedCrossRefGoogle Scholar
  8. 8.
    Hodges BL, Scheule RK. Hydrodynamic delivery of DNA. Expert Opin Biol Ther. 2003;3:911–8.PubMedCrossRefGoogle Scholar
  9. 9.
    Zhang G, Gao X, Song YK, Vollmer R, Stolz DB, Gasiorowski JZ, et al. Hydroporation as the mechanism of hydrodynamic delivery. Gene Ther. 2004;11:675–82.PubMedCrossRefGoogle Scholar
  10. 10.
    Suda T, Gao X, Stolz DB, Liu D. Structural impact of hydrodynamic injection on mouse liver. Gene Ther. 2007;14:129–37.PubMedGoogle Scholar
  11. 11.
    Kobayashi N, Nishikawa M, Hirata K, Takakura Y. Hydrodynamics-based procedure involves transient hyperpermeability in the hepatic cellular membrane: implication of a nonspecific process in efficient intracellular gene delivery. J Gene Med. 2004;6:584–92.PubMedCrossRefGoogle Scholar
  12. 12.
    Hagstrom JE, Hegge J, Zhang G, Noble M, Budker V, Lewis DL, et al. A facile nonviral method for delivering genes and siRNAs to skeletal muscle of mammalian limbs. Mol Ther. 2004;10:386–98.PubMedCrossRefGoogle Scholar
  13. 13.
    Zhang G, Budker V, Williams P, Subbotin V, Wolff JA. Efficient expression of naked DNA delivered intraarterially to limb muscles of nonhuman primates. Hum Gene Ther. 2001;12:427–38.PubMedCrossRefGoogle Scholar
  14. 14.
    Maruyama H, Higuchi N, Nishikawa Y, Kameda S, Iino N, Kazama JJ, et al. High-level expression of naked DNA delivered to rat liver via tail vein injection. J Gene Med. 2002;4:333–41.PubMedCrossRefGoogle Scholar
  15. 15.
    Eastman SJ, Baskin KM, Hodges BL, Chu G, Gates A, Dreusicke R, et al. Development of catheter-based procedures for transducing the isolated rabbit liver with plasmid DNA. Hum Gene Ther. 2002;13:2065–77.PubMedCrossRefGoogle Scholar
  16. 16.
    McCaffrey AP, Ohashi K, Meuse L, Shen S, Lancaster AM, Lukavsky PJ, et al. Determinants of hepatitis C translational initiation in vitro, in cultured cells and mice. Mol Ther. 2002;5:676–84.PubMedCrossRefGoogle Scholar
  17. 17.
    Zhang G, Song YK, Liu D. Long-term expression of human alpha1-antitrypsin gene in mouse liver achieved by intravenous administration of plasmid DNA using a hydrodynamics-based procedure. Gene Ther. 2000;7:1344–9.PubMedCrossRefGoogle Scholar
  18. 18.
    Stoll SM, Sclimenti CR, Baba EJ, Meuse L, Kay MA, Calos MP. Epstein-Barr virus/human vector provides high-level, long-term expression of alpha1-antitrypsin in mice. Mol Ther. 2001;4:122–9.PubMedCrossRefGoogle Scholar
  19. 19.
    Alino SF, Crespo A, Dasi F. Long-term therapeutic levels of human alpha-1 antitrypsin in plasma after hydrodynamic injection of nonviral DNA. Gene Ther. 2003;10:1672–9.PubMedCrossRefGoogle Scholar
  20. 20.
    Miao CH, Thompson AR, Loeb K, Ye X. Long-term and therapeutic-level hepatic gene expression of human factor IX after naked plasmid transfer in vivo. Mol Ther. 2001;3:947–57.PubMedCrossRefGoogle Scholar
  21. 21.
    Suda T, Liu D. Hydrodynamic gene delivery: its principles and applications. Mol Ther. 2007;15:2063–9.PubMedCrossRefGoogle Scholar
  22. 22.
    Magin-Lachmann C, Kotzamanis G, D’Aiuto L, Cooke H, Huxley C, Wagner E. In vitro and in vivo delivery of intact BAC DNA—comparison of different methods. J Gene Med. 2004;6:195–209.PubMedCrossRefGoogle Scholar
  23. 23.
    Chang J, Sigal LJ, Lerro A, Taylor J. Replication of the human hepatitis delta virus genome is initiated in mouse hepatocytes following intravenous injection of naked DNA or RNA sequences. J Virol. 2001;75:3469–73.PubMedCrossRefGoogle Scholar
  24. 24.
    Giladi H, Ketzinel-Gilad M, Rivkin L, Felig Y, Nussbaum O, Galun E. Small interfering RNA inhibits hepatitis B virus replication in mice. Mol Ther. 2003;8:769–76.PubMedCrossRefGoogle Scholar
  25. 25.
    Kobayashi N, Matsui Y, Kawase A, Hirata K, Miyagishi M, Taira K, et al. Vector-based in vivo RNA interference: dose- and time-dependent suppression of transgene expression. J Pharmacol Exp Ther. 2004;308:688–93.PubMedCrossRefGoogle Scholar
  26. 26.
    Layzer JM, McCaffrey AP, Tanner AK, Huang Z, Kay MA, Sullenger BA. In vivo activity of nuclease-resistant siRNAs. RNA. 2004;10:766–71.PubMedCrossRefGoogle Scholar
  27. 27.
    McCaffrey AP, Meuse L, Pham TT, Conklin DS, Hannon GJ, Kay MA. RNA interference in adult mice. Nature. 2002;418:38–9.PubMedCrossRefGoogle Scholar
  28. 28.
    McCaffrey AP, Meuse L, Karimi M, Contag CH, Kay MA. A potent and specific morpholino antisense inhibitor of hepatitis C translation in mice. Hepatology. 2003;38:503–8.PubMedCrossRefGoogle Scholar
  29. 29.
    Kobayashi N, Kuramoto T, Yamaoka K, Hashida M, Takakura Y. Hepatic uptake and gene expression mechanisms following intravenous administration of plasmid DNA by conventional and hydrodynamics-based procedures. J Pharmacol Exp Ther. 2001;297:853–60.PubMedGoogle Scholar
  30. 30.
    Nakata K, Tanaka Y, Nakano T, Adachi T, Tanaka H, Kaminuma T, et al. Nuclear receptor-mediated transcriptional regulation in Phase I, II, and III xenobiotic metabolizing systems. Drug Metab Pharmacokinet. 2006;21:437–57.PubMedCrossRefGoogle Scholar
  31. 31.
    Donato MT, Lahoz A, Castell JV, Gómez-Lechón MJ. Cell lines: a tool for in vitro drug metabolism studies. Curr Drug Metab. 2008;9:1–11.PubMedCrossRefGoogle Scholar
  32. 32.
    Al-Dosari MS, Knapp JE, Liu D. Activation of human CYP2C9 promoter and regulation by CAR and PXR in mouse liver. Mol Pharm. 2006;3:322–8.PubMedCrossRefGoogle Scholar
  33. 33.
    Schuetz E, Lan L, Yasuda K, Kim R, Kocarek TA, Schuetz J, et al. Development of a real-time in vivo transcription assay: application reveals pregnane X receptor-mediated induction of CYP3A4 by cancer chemotherapeutic agents. Mol Pharmacol. 2002;62:439–45.PubMedCrossRefGoogle Scholar
  34. 34.
    Tirona RG, Lee W, Leake BF, Lan LB, Cline CB, Lamba V, et al. The orphan nuclear receptor HNF4alpha determines PXR- and CAR-mediated xenobiotic induction of CYP3A4. Nat Med. 2003;9:220–4.PubMedCrossRefGoogle Scholar
  35. 35.
    Zhang W, Purchio A, Chen K, Burns SM, Contag CH, Contag PR. In vivo activation of the human CYP3A4 promoter in mouse liver and regulation by pregnane X receptors. Biochem Pharmacol. 2003;65:1889–96.PubMedGoogle Scholar
  36. 36.
    Merrell MD, Jackson JP, Augustine LM, Fisher CD, Slitt AL, Maher JM, et al. The Nrf2 activator oltipraz also activates the constitutive androstane receptor. Drug Metab Dispos. 2008;36:1716–21.PubMedCrossRefGoogle Scholar
  37. 37.
    Zelko I, Sueyoshi T, Kawamoto T, Moore R, Negishi M. The peptide near the C terminus regulates receptor CAR nuclear translocation induced by xenochemicals in mouse liver. Mol Cell Biol. 2001;21:2838–46.PubMedCrossRefGoogle Scholar
  38. 38.
    Sueyoshi T, Moore R, Pascussi JM, Negishi M. Direct expression of fluorescent protein-tagged nuclear receptor CAR in mouse liver. Methods Enzymol. 2002;357:205–13.PubMedCrossRefGoogle Scholar
  39. 39.
    Sueyoshi T, Moore R, Sugatani J, Matsumura Y, Negishi M. PPP1R16A, the membrane subunit of protein phosphatase 1beta, signals nuclear translocation of the nuclear receptor constitutive active/androstane receptor. Mol Pharmacol. 2008;73:1113–21.PubMedCrossRefGoogle Scholar
  40. 40.
    Meng Z, Wang Y, Wang L, Jin W, Liu N, Pan H, et al. FXR regulates liver repair after CCl4-induced toxic injury. Mol Endocrinol. 2010;24:886–97.PubMedCrossRefGoogle Scholar
  41. 41.
    Shepard CW, Simard EP, Finelli L, Fiore AE, Bell BP. Hepatitis B virus infection: epidemiology and vaccination. Epidemiol Rev. 2006;28:112–25.PubMedCrossRefGoogle Scholar
  42. 42.
    Zanetti AR, Van Damme P, Shouval D. The global impact of vaccination against hepatitis B: A historical overview. Vaccine. 2008;26:6266–73.PubMedCrossRefGoogle Scholar
  43. 43.
    Bertoni R, Sette A, Sidney J, Guidotti LG, Shapiro M, Purcell R, et al. Human class I supertypes and CTL repertoires extend to chimpanzees. J Immunol. 1998;161:4447–55.PubMedGoogle Scholar
  44. 44.
    Will H, Cattaneo R, Koch HG, Darai G, Schaller H, Schellekens H, et al. Cloned HBV DNA causes hepatitis in chimpanzees. Nature. 1982;299:740–2.PubMedCrossRefGoogle Scholar
  45. 45.
    Yang PL, Althage A, Chung J, Chisari FV. Hydrodynamic injection of viral DNA: a mouse model of acute hepatitis B virus infection. Proc Natl Acad Sci USA. 2002;99:13825–30.PubMedCrossRefGoogle Scholar
  46. 46.
    Suzuki T, Takehara T, Ohkawa K, Ishida H, Jinushi M, Miyagi T, et al. Intravenous injection of naked plasmid DNA encoding hepatitis B virus (HBV) produces HBV and induces humoral immune response in mice. Biochem Biophys Res Commun. 2003;300:784–8.PubMedCrossRefGoogle Scholar
  47. 47.
    Huang LR, Wu HL, Chen PJ, Chen DS. An immunocompetent mouse model for the tolerance of human chronic hepatitis B virus infection. Proc Natl Acad Sci USA. 2006;103:17862–7.PubMedCrossRefGoogle Scholar
  48. 48.
    Wang Q, Contag CH, Ilves H, Johnston BH, Kaspar RL. Small hairpin RNAs efficiently inhibit hepatitis C IRES–mediated gene expression in human tissue culture cells and a mouse model. Mol Ther. 2005;12:562–8.PubMedCrossRefGoogle Scholar
  49. 49.
    Kim SI, Shin D, Lee H, Ahn B, Yoon Y, Kim M. Targeted delivery of siRNA against hepatitis C virus by apolipoprotein A-I-bound cationic liposomes. J Hepat. 2009;50:479–88.CrossRefGoogle Scholar
  50. 50.
    Nakamura G, Maruyama H, Ishii S, Shimotori M, Kameda S, Kono T, et al. Naked plasmid DNA-based alpha-galactosidase A gene transfer partially reduces systemic accumulation of globotriaosylceramide in Fabry mice. Mol Biotechnol. 2008;38:109–19.PubMedCrossRefGoogle Scholar
  51. 51.
    Chen L, Woo SL. Complete and persistent phenotypic correction of phenylketonuria in mice by site-specific genome integration of murine phenylalanine hydroxylase cDNA. Proc Natl Acad Sci USA. 2005;102:15581–6.PubMedCrossRefGoogle Scholar
  52. 52.
    Sondergaard M, Dagnaes-Hansen F, Flyvbjerg A, Jensen TG. Normalization of growth in hypophysectomized mice using hydrodynamic transfer of the human growth hormone gene. Am J Physiol Endocrinol Metab. 2003;285:E427–32.PubMedGoogle Scholar
  53. 53.
    Takakusaki Y, Hisayasu S, Hirai Y, Shimada T. Coexpression of formylglycine-generating enzyme is essential for synthesis and secretion of functional arylsulfatase A in a mouse model of metachromatic leukodystrophy. Hum Gene Ther. 2005;16:929–36.PubMedCrossRefGoogle Scholar
  54. 54.
    Camassola M, Braga LM, Delgado-Canedo A, Dalberto TP, Matte U, Burin M. Nonviral in vivo gene transfer in the mucopolysaccharidosis I murine model. J Inherit Metab Dis. 2005;28:1035–43.PubMedCrossRefGoogle Scholar
  55. 55.
    Holm DA, Dagnaes-Hansen F, Simonsen H, Gregersen N, Bolund L, Jensen TG. Expression of short-chain acyl-CoA dehydrogenase (SCAD) proteins in the liver of SCAD deficient mice after hydrodynamic gene transfer. Mol Genet Metab. 2003;78:250–8.PubMedCrossRefGoogle Scholar
  56. 56.
    He CX, Shi D, Wu WJ, Ding YF, Feng DM, Lu B. Insulin expression in livers of diabetic mice mediated by hydrodynamics-based administration. World J Gastroenterol. 2004;10:567–72.PubMedGoogle Scholar
  57. 57.
    Dai C, Yang J, Bastacky S, Xia J, Li Y, Liu Y. Intravenous administration of hepatocyte growth factor gene ameliorates diabetic nephropathy in mice. J Am Soc Nephrol. 2004;15:2637–47.PubMedCrossRefGoogle Scholar
  58. 58.
    Gonzalez Muniesa P, Milagro FI, Campion J, Martinez JA. Reduction in energy efficiency induced by expression of the uncoupling protein, UCP1, in mouse liver mitochondria. Int J Mol Med. 2006;17:591–7.PubMedGoogle Scholar
  59. 59.
    Jiang J, Yamato E, Miyazaki J. Long-term control of food intake and body weight by hydrodynamics-based delivery of plasmid DNA encoding leptin or CNTF. J Gene Med. 2003;5:977–83.PubMedCrossRefGoogle Scholar
  60. 60.
    Chang H, Hanawa H, Liu H, Yoshida T, Hayashi M, Watanabe R. Hydrodynamic-based delivery of an interleukin-22-Ig fusion gene ameliorates experimental autoimmune myocarditis in rats. J Immunol. 2006;177:3635–43.PubMedGoogle Scholar
  61. 61.
    Elnaggar R, Hanawa H, Liu H, Yoshida T, Hayashi M, Watanabe R. The effect of hydrodynamics-based delivery of an IL-13-Ig fusion gene for experimental autoimmune myocarditis in rats and its possible mechanism. Eur J Immunol. 2005;35:1995–2005.PubMedCrossRefGoogle Scholar
  62. 62.
    Liu H, Hanawa H, Yoshida T, Elnaggar R, Hayashi M, Watanabe R. Effect of hydrodynamics-based gene delivery of plasmid DNA encoding interleukin-1 receptor antagonist-Ig for treatment of rat autoimmune myocarditis: possible mechanism for lymphocytes and noncardiac cells. Circulation. 2005;111:1593–600.PubMedCrossRefGoogle Scholar
  63. 63.
    Higuchi N, Maruyama H, Kuroda T, Kameda S, Iino N, Kawachi H. Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar-Kyoto rats. Gene Ther. 2003;10:1297–310.PubMedCrossRefGoogle Scholar
  64. 64.
    Fu AL, Wang YX, Sun MJ. Naked DNA prevents soman intoxication. Biochem Biophys Res Commun. 2005;328:901–5.PubMedCrossRefGoogle Scholar
  65. 65.
    Miki Y, Maruyama S, Liu D, Kobayashi T, Sato F, Shimizu H. In vivo gene transfer of endo-beta-galactosidase C removes alphaGal antigen on erythrocytes and endothelial cells of the organs. Xenotransplantation. 2004;11:444–51.PubMedCrossRefGoogle Scholar
  66. 66.
    Inoue S, Hakamata Y, Kaneko M, Kobayashi E. Gene therapy for organ grafts using rapid injection of naked DNA: application to the rat liver. Transplantation. 2004;77:997–1003.PubMedCrossRefGoogle Scholar
  67. 67.
    Wang CH, Liang CL, Huang LT, Liu JK, Hung PH, Sun A. Single intravenous injection of naked plasmid DNA encoding erythropoietin provides neuroprotection in hypoxia-ischemia rats. Biochem Biophys Res Commun. 2004;314:1064–71.PubMedCrossRefGoogle Scholar
  68. 68.
    Zeini M, Hortelano S, Traves PG, Gomez Valades AG, Pujol A, Perales JC. Assessment of a dual regulatory role for NO in liver regeneration after partial hepatectomy: protection against apoptosis and retardation of hepatocyte proliferation. FASEB J. 2005;19:995–7.PubMedGoogle Scholar
  69. 69.
    Barnett FH, Scharer-Schuksz M, Wood M, Yu X, Wagner TE, Friedlander M. Intra-arterial delivery of endostatin gene to brain tumors prolongs survival and alters tumor vessel ultrastructure. Gene Ther. 2004;11:1283–9.PubMedCrossRefGoogle Scholar
  70. 70.
    Tada M, Hatano E, Taura K, Nitta T, Koizumi N, Ikai I. High volume hydrodynamic injection of plasmid DNA via the hepatic artery results in a high level of gene expression in rat hepatocellular carcinoma induced by diethylnitrosamine. J Gene Med. 2006;8:1018–26.PubMedCrossRefGoogle Scholar
  71. 71.
    Kobayashi N, Kuramoto T, Chen S, Watanabe Y, Takakura Y. Therapeutic effect of intravenous interferon gene delivery with naked plasmid DNA in murine metastasis models. Mol Ther. 2002;6:737–44.PubMedCrossRefGoogle Scholar
  72. 72.
    Yazawa H, Murakami T, Li HM, Back T, Kurosaka K, Suzuki Y. Hydrodynamics-based gene delivery of naked DNA encoding fetal liver kinase-1 gene effectively suppresses the growth of pre-existing tumors. Cancer Gene Ther. 2006;13:993–1001.PubMedCrossRefGoogle Scholar
  73. 73.
    Sato A, Ohtsuki M, Hata M, Kobayashi E, Murakami T. Antitumor activity of IFN-lambda in murine tumor models. J Immunol. 2006;176:7686–94.PubMedGoogle Scholar
  74. 74.
    Kim KS, Park YS. Antitumor effects of angiostatin K1-3 and endostatin genes coadministered by the hydrodynamics-based transfection method. Oncol Res. 2005;15:343–50.PubMedGoogle Scholar
  75. 75.
    Takehara T, Uemura A, Tatsumi T, Suzuki T, Kimura R, Shiotani A. Natural killer cell-mediated ablation of metastatic liver tumors by hydrodynamic injection of IFNalpha gene to mice. Int J Cancer. 2007;120:1252–60.PubMedCrossRefGoogle Scholar
  76. 76.
    Yonenaga Y, Mori A, Fujimoto A, Nagayama S, Tachibana T, Onodera H. The administration of naked plasmid DNA into the liver induces antitumor innate immunity in a murine liver metastasis model. J Gene Med. 2007;9:299–307.PubMedCrossRefGoogle Scholar
  77. 77.
    Kitajima M, Tsuyama Y, Miyano-Kurosaki N, Takaku H. Anti-tumor effect of intravenous TNFalpha gene delivery naked plasmid DNA using a hydrodynamics-based procedure. Nucleosides Nucleotides Nucleic Acids. 2005;24:647–50.PubMedCrossRefGoogle Scholar
  78. 78.
    Chen HW, Lee YP, Chung YF, Shih YV, Tsai JP, Tao MH. Inducing long-term survival with lasting anti-tumor immunity in treating B cell lymphoma by a combined dendritic cell-based and hydrodynamic plasmid-encoding IL-12 gene therapy. Int Immunol. 2003;15:427–35.PubMedCrossRefGoogle Scholar
  79. 79.
    Wang G, Tschoi M, Spolski R, Lou Y, Ozaki K, Feng C. In vivo antitumor activity of interleukin 21 mediated by natural killer cells. Cancer Res. 2003;63:9016–22.PubMedGoogle Scholar
  80. 80.
    Wen J, Matsumoto K, Taniura N, Tomioka D, Nakamura T. Hepatic gene expression of NK4, an HGF-antagonist/angiogenesis inhibitor, suppresses liver metastasis and invasive growth of colon cancer in mice. Cancer Gene Ther. 2004;11:419–30.PubMedCrossRefGoogle Scholar
  81. 81.
    Ortaldo JR, Winkler-Pickett RT, Bere EW, Watanabe Jr M, Murphy MJ, Wiltrout RH. In vivo hydrodynamic delivery of cDNA encoding IL-2: rapid, sustained redistribution, activation of mouse NK cells, and therapeutic potential in the absence of NKT cells. J Immunol. 2005;175:693–9.PubMedGoogle Scholar
  82. 82.
    Neal ZC, Bates MK, Albertini MR, Herweijer H. Hydrodynamic limb vein delivery of a xenogeneic DNA cancer vaccine effectively induces antitumor immunity. Mol Ther. 2007;15:422–30.PubMedCrossRefGoogle Scholar
  83. 83.
    Tward AD, Jones KD, Yant S, Cheung ST, Fan ST, Chen X, et al. Distinct pathways of genomic progression to benign and malignant tumors of the liver. Proc Natl Acad Sci USA. 2007;104:14771–6.PubMedCrossRefGoogle Scholar
  84. 84.
    Collins MA, Shaw I, Billington DC. Driving drug discovery and patient therapy via the encapsulation and fusion of knowledge. Drug Des Discov. 1999;16:181–94.PubMedGoogle Scholar
  85. 85.
    Suda T, Suda K, Liu D. Computer-assisted hydrodynamic gene delivery. Mol Ther. 2008;16:1098–104.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of Pharmaceutical SciencesUniversity of Pittsburgh School of PharmacyPittsburghUSA

Personalised recommendations