Abstract
Background
Hepatocyte growth factor/scatter factor (HGF/SF) is a pleiotropic cytokine with mitogenic, motogenic and morphogenic effects for a wide variety of cells. Previous studies have reported that the in vivo infusion in normal, untreated mice of recombinant HGF results in low levels of DNA synthesis and liver proliferation. In this study, we examined whether liver regeneration could be obtained by the in vivo injection of a recombinant adenoviral vector encoding human HGF (Ad.CMV.rhHGF) in normal, intact mice.
Materials and Methods
C57BL/6 mice were infused intravenously with doses increasing from 1 to 4 × 1011 particles of the recombinant human HGF (rhHGF) adenoviral vector or with a control virus encoding Escherichia coli β-galactosidase (Ad.CMV.lacZ). At day 5, mice were sacrificed and evaluated for the presence of hepatocyte mitogenesis and liver regeneration (5-bromo-2′-deoxyuridine (BrdU) assays and liver weight determination) and for the presence of liver damage (serum alanine amino-transferase (ALT) measurements and TUNEL assays).
Results
In vivo administration of rhHGF stimulated DNA synthesis of hepatocytes and liver weight in a dose-dependent fashion. The maximal effect was seen after the infusion of 3 × 1011 particles which resulted at day 5 in >130% increase in relative liver mass with little cytopathic effect. In contrast, administration of the lacZ adenoviral vector caused little hepatocyte replication, but induced high levels of serum ALT (∼3 times higher than the rhHGF vector) and significant apoptotic cell death.
Conclusions
This study shows that a single injection of Ad.CMV.rhHGF alone is able to induce in vivo and in a very short period of time, robust DNA synthesis and liver proliferation in normal mice without liver injury or partial hepatectomy. This recombinant adenoviral vector has a lower toxicity than the control lacZ adenovirus. This suggests that HGF may have a protective effect against adenovirus-induced pathology.
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References
Matsumoto K, Nakamura N. (1996) Emerging multipotent aspects of hepatocyte growth factor. J Biochem. (Tokyo) 119: 591–600.
Stoker M, Gherardi E, Perryman M, Gray J. (1987) Scatter factor is a fibroblast-derived modulator of epithelial cell mobility. Nature 327: 239–242.
Naldini L, Weidner KM, Vigna E, et al. (1991) Scatter factor and hepatocyte growth factor are indistinguishable ligands for the MET receptor. EMBO J. 10: 2867–2878.
Nakamura T, Nawa K, Ichihara A, Kaise N, Nishino T. (1987) Purification and subunit structure of hepatocyte growth factor from rat platelets. FEBS Lett. 224: 311–316.
Rubin JS, Bottaro DP, Aaronson SA. (1993) Hepatocyte growth factor/scatter factor and its receptor, the c-met proto-oncogene product. Biochim. Biophys. Acta. 1155: 357–371.
Gohda E, Tsubouchi H, Nakayama H, et al. (1988) Purification and partial characterization of hepatocyte growth factor from plasma of a patient with fulminant hepatic failure. J. Clin. Invest. 81: 414–419.
Asami O, Ihara I, Shimidzu N, Shimizu S, Tomita Y, Ichihara A, Nakamura T. (1991) Purification and characterization of hepatocyte growth factor from injured liver of carbon tetrachloridetreated rats. J. Biochem. (Tokyo) 109: 8–13.
Selden C, Johnstone R, Darby H, Gupta S, Hodgson HJ. (1986) Human serum does contain a high molecular weight hepatocyte growth factor: studies pre- and post-hepatic resection. Biochem. Biophys. Res. Commun. 139: 361–366.
Nakamura T, Nishizawa T, Hagiya M, et al. (1989) Molecular cloning and expression of human hepatocyte growth factor. Nature 342: 440–443.
Miyazawa K, Tsubouchi H, Naka D, et al. (1989) Molecular cloning and sequence analysis of cDNA for human hepatocyte growth factor. Biochem. Biophys. Res. Commun. 163: 967–973.
Tashiro K, Hagiya M, Nishizawa T, et al. (1990) Deduced primary structure of rat hepatocyte growth factor and expression of the mRNA in rat tissues. Proc. Natl. Acad. Sci. U.S.A. 87: 3200–3204.
Okajima A, Miyazawa K, Kitamura N. (1990) Primary structure of rat hepatocyte growth factor and induction of its mRNA during liver regeneration following hepatic injury. Eur. J. Biochem. 193: 375–381.
Liu Y, Michalopoulos GK, Zarnegar R. (1993) Molecular cloning and characterization of cDNA encoding mouse hepatocyte growth factor. Biochim. Biophys. Acta. 1216: 299–303.
Patijn GA, Lieber A, Schowalter DB, Schwall R, Kay MA. (1998) Hepatocyte growth factor induces hepatocyte proliferation in vivo and allows for efficient retroviral-mediated gene transfer in mice. Hepatology 28: 707–716.
Liu M-L, Mars WM, Zarnegar R, Michalopoulos GK. (1994) Collagenase pretreatment and the mitogenic effects of hepatocyte growth factor and transforming growth factor-α in adult rat liver. Hepatology 19: 1521–1527.
Ye X, Robinson MB, Batshaw ML, Furth EE, Smith I, Wilson JM. (1996) Prolonged metabolic correction in adult ornithine transcarbamylase-deficient mice with adenoviral vectors. J. Biol. Chem. 1996 271: 3639–3646.
Kozarsky KF, McKinley DR, Austin LL, Raper SE, Stratford-Perricaudet LD, Wilson JM. (1994) In vivo correction of low density lipoprotein receptor deficiency in the Watanabe heritable hyperlipidemic rabbit with recombinant adenoviruses. J. Biol. Chem. 269: 13695–13702.
Yang Y, Ertl HC, Wilson JM. (1994) MHC class I-restricted cytotoxic T lymphocytes to viral antigens destroy hepatocytes in mice infected with E1-deleted recombinant adenoviruses. Immunity 1: 433–442.
Yang Y, Nunes FA, Berencsi K, Furth EE, Gonczol E, Wilson JM. (1994) Cellular immunity to viral antigens limits El-deleted adenoviruses for gene therapy. Proc. Natl. Acad. Sci. U.S.A. 91: 4407–4411.
Jooss K, Yang Y, Wilson JM. (1996). Cyclophosphamide diminishes inflammation and prolongs transgene expression following delivery of adenoviral vectors to mouse liver and lung. Hum. Gene Ther. 7: 1555–1566.
Zarnegar R, Michalopoulos G. (1989) Purification and biological characterization of human hepatopoietin A, a polypeptide growth factor for hepatocytes. Cancer Res. 49: 3314–3320.
Nakamura T, Nawa K, Ichihara A. (1984) Partial purification and characterization of hepatocyte growth factor from serum of hepatectomized rats. Biochem. Biophys. Res. Commun. 122: 1450–1459.
Lindroos PM, Zarnegar R, Michalopoulos GK. (1991) Hepatocyte growth factor (hepatopoietin A) rapidly increases in plasma before DNA synthesis and liver regeneration stimulated by partial hepatectomy and carbon tetrachloride administration. Hepatology 13: 743–750.
Kinoshita T, Tashiro K, Nakamura T. (1989) Marked increase of HGF mRNA in non-parenchymal liver cells of rats treated with hepatotoxins. Biochem. Biophys. Res. Commun. 165: 1229–1234.
Shiota G, Wang TC, Nakamura T, Schmidt EV. (1994) Hepatocyte growth factor in transgenic mice: effects on hepatocyte growth, liver regeneration and gene expression. Hepatology 19: 962–972.
Webber EM, Godowski PJ, Fausto N. (1994) In vivo response of hepatocytes to growth factors requires an initial priming stimulus. Hepatology 19: 489–497.
Ishii T, Sato M, Sudo K, et al. (1995) Hepatocyte growth factor stimulates liver regeneration and elevates blood protein level in normal and partially hepatectomized rats. J. Biochem. (Tokyo) 117: 1105–1112.
Fujiwara K, Nagoshi S, Ohno A, et al. (1993) Stimulation of liver growth by exogenous human hepatocyte growth factor in normal and partially hepatectomized rats. Hepatology 18: 1443–1449.
Zioncheck TF, Richardson L, DeGuzman GG, Modi NB, Hansen SE, Godowski PJ. (1994) The pharmacokinetics, tissue localization, and metabolic processing of recombinant human hepatocyte growth factor after intravenous administration in rats. Endocrinology 134: 1879–1887.
Liu KX, Kato Y, Narukawa M, et al. (1992). Importance of the liver in plasma clearance of hepatocyte growth factors in rats. Am. J. Physiol. 263: G642–G649.
Kobayashi Y, Hamanoue M, Ueno S, et al. (1996) Induction of hepatocyte growth by intraportal infusion of HGF into beagle dogs. Biochem. Biophys. Res. Commun. 220: 7–12.
Kaido T, Yamaoka S, Tanaka J, et al. (1996) Continuous HGF supply from HGF-expressing fibroblasts transplanted into spleen prevents CCl4-induced acute liver injury in rats. Biochem. Biophys. Res. Commun. 218: 1–5.
Kaido T, Yamaoka S, Seto S, et al. (1997) Continuous hepatocyte growth factor supply prevents lipopolysaccharide-induced liver injury in rats. FEBS Lett. 411: 378–382.
Ueki T, Kaneda Y, Tsutsui H, et al. (1999) Hepatocyte growth factor gene therapy of liver cirrhosis in rats. Nat. Med. 5: 226–230.
Fan S, Wang JA, Yuan RQ, Rockwell S, Andres J, Zlatapolskiy A, Goldberg ID, Rosen EM. (1998) Scatter factor protects epithelial and carcinoma cells against apoptosis in-duced by DNA-damaging agents. Oncogene 17: 131–141.
Morita M, Watanabe Y, Akaike T. (1995) Protective effect of hepatocyte growth factor on interferon-gamma-induced cytotoxicity in mouse hepatocytes. Hepatology 21: 1585–1593.
Yo Y, Morishita R, Nakamura S, et al. (1998) Potential role of hepatocyte growth factor in the maintenance of renal structure: anti-apoptotic action of HGF on epithelial cells. Kidney Int. 54: 1128–1138.
Liu Y, Sun AM, Dworkin LD. (1998) Hepatocyte growth factor protects renal epithelial cells from apoptotic cell death. Biochem. Biophys. Res. Commun. 246: 821–826.
Gao C, Jokerst R, Gondipalli P, Cai SR, Kennedy S, Ponder KP. (1999) Intramuscular injection of an adenoviral vector expressing hepatocyte growth factor facilitates hepatic transduction with a retroviral vector in mice. Hum. Gene Ther. 10: 911–922.
Acknowledgments
Support from the Clinical Pathology Core of the Institute for Human Gene Therapy was appreciated. This work was supported by grants from the NIH (NICHD P01-AD32649-05, NIDDK P30-DK47757-06, and NHLBI P01-HL59407-01) and Genovo, Inc., a biotechnology company founded by Dr. Wilson.
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Phaneuf, D., Chen, SJ. & Wilson, J.M. Intravenous Injection of an Adenovirus Encoding Hepatocyte Growth Factor Results in Liver Growth and Has a Protective Effect Against Apoptosis. Mol Med 6, 96–103 (2000). https://doi.org/10.1007/BF03401777
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DOI: https://doi.org/10.1007/BF03401777