Low-dose simultaneous delivery of adenovirus encoding hepatocyte growth factor and vascular endothelial growth factor in dogs enhances liver proliferation without systemic growth factor elevation
Hussein M. Atta,
Ayman Al-Hendy,
Salama A. Salama,
Olfat G. Shaker,
Olfat A. Hammam,
Hussein M. Atta
Department of Surgery, Faculty of Medicine, Minia University, Minia, Egypt
Search for more papers by this authorAyman Al-Hendy
Department of Obstetric and Gynecology, Meharry Medical College, Nashville, TN, USA
Search for more papers by this authorSalama A. Salama
Department of Obstetrics and Gynecology, University of Texas Medical Branch at Galveston, Galveston, TX, USA
Search for more papers by this authorOlfat G. Shaker
Department of Medical Biochemistry, Cairo University, Cairo, Egypt
Search for more papers by this authorOlfat A. Hammam
Department of Pathology, Theodor Bilharz Research Institute, Giza, Egypt
Search for more papers by this authorHussein M. Atta,
Ayman Al-Hendy,
Salama A. Salama,
Olfat G. Shaker,
Olfat A. Hammam,
Hussein M. Atta
Department of Surgery, Faculty of Medicine, Minia University, Minia, Egypt
Search for more papers by this authorAyman Al-Hendy
Department of Obstetric and Gynecology, Meharry Medical College, Nashville, TN, USA
Search for more papers by this authorSalama A. Salama
Department of Obstetrics and Gynecology, University of Texas Medical Branch at Galveston, Galveston, TX, USA
Search for more papers by this authorOlfat G. Shaker
Department of Medical Biochemistry, Cairo University, Cairo, Egypt
Search for more papers by this authorOlfat A. Hammam
Department of Pathology, Theodor Bilharz Research Institute, Giza, Egypt
Search for more papers by this authorCorrespondence
Hussein M. Atta, MD, PhD, Department of Surgery, Faculty of Medicine, Minia University, Misr-Aswan Road, El-Minia 61519, EgyptTel: +202 22917077Fax: +2068 2342502e-mail: [email protected]
Abstract
Background: Hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) gene transfer proved to enhance liver regeneration. However, elevation of their plasma levels may induce potentially serious distant effects such as tumorigenesis or proliferative retinopathy.
Aims: This study was performed to examine whether simultaneous administration of low-dose adenovirus encoding HGF and VEGF genes in dogs will stimulate liver proliferation but without inducing liver toxicity or systemic elevation of HGF and VEGF levels.
Methods: Adult dogs received an intravenous injection of low-dose adenoviral vectors encoding human HGF and VEGF (HGF/VEGF), β-galactosidase (lacZ) or phosphate-buffered saline (PBS). Liver proliferation was measured using the proliferating cell nuclear antigen (PCNA) immunostaining labelling index. HGF and VEGF plasma concentrations and transaminases were repeatedly measured. Transgene expression was evaluated using reverse-transcription polymerase chain reaction.
Results: Human HGF and VEGF expressions were detected only in the liver of HGF/VEGF dogs at day 2 after injection but declined at sacrifice (day 7). No expression was detected in the liver of the lacZ or PBS groups. Plasma levels of HGF and VEGF were not statistically different from those in the lacZ group (P=0.81, P=0.22 respectively). The PCNA labelling index was five-fold higher in the HGF/VEGF group compared with the lacZ group (P<0.01). No immunostaining was detected in the PBS group. Transaminases were only elevated (P<0.01) in the lacZ group compared with the other groups.
Conclusions: We showed that simultaneous administration of low-dose adenoviral vectors encoding human HGF and VEGF genes can induce transgene expression and liver proliferation without liver toxicity or systemic growth factor elevation.
References
- 1 Phaneuf D, Chen SJ, Wilson JM. Intravenous injection of an adenovirus encoding hepatocyte growth factor results in liver growth and has a protective effect against apoptosis. Mol Med 2000; 6: 96–103.
- 2 Patijn GA, Lieber A, Schowalter DB, et al. Hepatocyte growth factor induces hepatocyte proliferation in vivo and allows for efficient retroviral-mediated gene transfer in mice. Hepatology 1998; 28: 707–16.
- 3 Shiota G, Kunisada T, Oyama K, et al. In vivo transfer of hepatocyte growth factor gene accelerates proliferation of hepatic oval cells in a 2-acetylaminofluorene/partial hepatectomy model in rats. FEBS Lett 2000; 470: 325–30.
- 4 Nomi T, Shiota G, Isono M, et al. Adenovirus-mediated hepatocyte growth factor gene transfer prevents lethal liver failure in rats. Biochem Biophys Res Commun 2000; 278: 338–43.
- 5 Ueki T, Kaneda Y, Tsutsui H, et al. Hepatocyte growth factor gene therapy of liver cirrhosis in rats. Nat Med 1999; 5: 226–30.
- 6 Yu Q, Que LG, Rockey DC. Adenovirus-mediated gene transfer to nonparenchymal cells in normal and injured liver. Am J Physiol Gastrointest Liver Physiol 2002; 282: G565–72.
- 7 Redaelli CA, Semela D, Carrick FE, et al. Effect of vascular endothelial growth factor on functional recovery after hepatectomy in lean and obese mice. J Hepatol 2004; 40: 305–12.
- 8 Fausto N, Riehle KJ. Mechanisms of liver regeneration and their clinical implications. J Hepatobiliary Pancreat Surg 2005; 12: 181–9.
- 9 Michalopoulos GK, DeFrances MC. Liver regeneration. Science 1997; 276: 60–6.
- 10 Tomiya T, Ogata I, Yamaoka M, et al. The mitogenic activity of hepatocyte growth factor on rat hepatocytes is dependent upon endogenous transforming growth factor-α. Am J Pathol 2000; 157: 1693–701.
- 11 Harrison P, Gove C, Bomford A. Hepatic expression of hepatocyte growth factor gene mRNA in acute liver failure. Dig Dis Sci 2000; 45: 1913–20.
- 12 Kosai K, Matsumoto K, Funakoshi H, Nakamura T. Hepatocyte growth factor prevents endotoxin-induced lethal hepatic failure in mice. Hepatology 1999; 30: 151–9.
- 13 Seto S, Kaido T, Yamaoka S, et al. Hepatocyte growth factor prevents lipopolysaccharide-induced hepatic sinusoidal endothelial cell injury and intrasinusoidal fibrin deposition in rats. J Surg Res 1998; 80: 194–9.
- 14 Kaido T, Yamaoka S, Seto S, et al. Continuous hepatocyte growth factor supply prevents lipopolysaccharide-induced liver injury in rats. FEBS Lett 1997; 411: 378–82.
- 15 Shiota G, Wang TC, Nakamura T, Schmidt EV. Hepatocyte growth factor in transgenic mice: effect on hepatocyte growth, liver regeneration and gene expression. Hepatology 1994; 19: 962–72.
- 16 Kaido T, Yoshikawa A, Seto S, et al. Hepatocyte growth factor supply accelerates compensatory hypertrophy caused by portal branch ligation in normal and jaundiced rats. J Surg Res 1999; 85: 115–9.
- 17 Fausto N. Liver regeneration. J Hepatol 2000; 32 (Suppl.): 19–31.
- 18 LeCouter J, Moritz DR, Li B, et al. Angiogenesis-independent endothelial protection of liver: role of VRGFR-1. Science 2003; 299: 890–3.
- 19 Ramadori G, Neubauer K, Odenthal M, et al. The gene of hepatocyte growth factor is expressed in fat-storing cells of rat liver and is downregulated during cell growth and by transforming growth factor-beta. Biochem Biophys Res Commun 1992; 183: 739–42.
- 20 Borowiak M, Garratt AN, Wustefeld T, et al. Met provides essential signals for liver regeneration. Proc Natl Acad Sci USA 2004; 101: 10608–13.
- 21 Bockhorn M, Goralski M, Prokofiev D, et al. VEGF is important for early liver regeneration after partial hepatectomy. J Surg Res 2007; 138: 291–9.
- 22 Marino G, Piazzese E, Gruttadauria S, et al. New model of liver regeneration induced through use of vascular endothelial growth factor. Transpl Proc 2006; 38: 1193–4.
- 23 Furnus CC, Inda AM, Andrini LB, et al. Chronobiology of the proliferative events related to angiogenesis in mice liver regeneration after partial hepatectomy. Cell Biol Int 2003; 27: 383–6.
- 24 Kraizer Y, Mawasi N, Seagal J, et al. Vascular endothelial growth factor and angiopoietin in liver regeneration. Biochem Biophys Res Commun 2001; 287: 209–15.
- 25 Assy N, Spira G, Paizi M, et al. Effect of vascular endothelial growth factor on hepatic regenerative activity following partial hepatectomy in rats. J Hepatol 1999; 30: 911–5.
- 26 Ferrara N, Davis-Smyth T. The biology of vascular endothelial growth factor. Endocrine Rev 1997; 18: 4–25.
- 27 Oe H, Kaido T, Furuyama H, et al. Simultaneous transfer of vascular endothelial growth factor and hepatocytes growth factor genes effectively promotes liver regeneration after hepatectomy in cirrhotic rats. Hepatogastroenterology 2004; 51: 1641–7.
- 28 Sato T, El-Assal ON, Ono T, et al. Sinusoidal endothelial cell proliferation and expression of angiopoietin/Tie family in regenerating rat liver. J Hepatol 2001; 34: 690–8.
- 29 Ross MA, Sander CM, Kleeb TB, et al. Spatiotemporal expression of angiogenesis growth factor receptors during the revascularization of regenerating rat liver. Hepatology 2001; 34: 1135–48.
- 30 Taniguchi E, Sakisaka S, Matsuo K, et al. Expression and role of vascular endothelial growth factor in liver regeneration after partial hepatectomy in rats. J Histochem Cytochem 2001; 49: 121–9.
- 31 Shimizu H, Miyazaki M, Wakabayashi Y, et al. Vascular endothelial growth factor secreted by replicating hepatocytes induces sinusoidal endothelial cell proliferation during regeneration after partial hepatectomy in rats. J Hepatol 2001; 34: 683–9.
- 32 Davidson AJ, Zon LI. Love, honor, and protect (your liver). Science 2003; 299: 835–7.
- 33 Shams N, Ianchulev T. Role of vascular endothelial growth factor in ocular angiogenesis. Ophthalmol Clin North Am 2006; 19: 335–44.
- 34 Wu F, Wu L, Zheng S, et al. The clinical value of hepatocyte growth factor and its receptor-c-met for liver cancer patients with hepatectomy. Dig Liver Dis 2006; 38: 490–7.
- 35 Morral N, O'Neal WK, Rice K, et al. Lethal toxicity, severe endothelial injury, and a threshold effect with high doses of an adenoviral vector in baboons. Hum Gene Ther 2002; 13: 143–54.
- 36 Nunes FA, Furth EE, Wilson JM, Raper SE. Gene transfer into the liver of nonhuman primates with E1-deleted recombinant adenoviral vectors: safety of readministration. Hum Gene Ther 1999; 10: 2515–26.
- 37 Tao N, Gao GP, Parr M, et al. Sequestration of adenoviral vector by Kupffer cells leads to a nonlinear dose response of transduction in liver. Mol Ther 2001; 3: 28–35.
- 38 Ehrhardt A, Xu H, Dillow AM, et al. A gene-deleted adenoviral vector results in phenotypic correction of canine hemophilia B without liver toxicity or thrombocytopenia. Blood 2003; 102: 2403–11.
- 39 Chuah MK, Schiedner G, Thorrez L, et al. Therapeutic factor VIII levels and negligible toxicity in mouse and dog models of hemophilia A following gene therapy with high-capacity adenoviral vectors. Blood 2003; 101: 1734–43.
- 40 Olson H, Betton G, Robinson D, et al. Concordance of the toxicity of pharmaceuticals in humans and in animals. Regul Toxicol Pharmacol 2000; 32: 56–67.
- 41 Raja NU, Holman DH, Wang D, et al. Induction of bivalent immune responses by expression of dengue virus type 1 and type 2 antigens from a single complex adenoviral vector. Am J Trop Med Hyg 2007; 76: 743–51.
- 42 Miyake M, Saze K, Yaguchi T, et al. Canine hepatocyte growth factor: molecular cloning and characterization of the recombinant protein. Vet Immunol Immunopathol 2003; 95: 135–43.
- 43 Oe H, Kaido T, Mori A, et al. Hepatocyte growth factor as well as vascular endothelial growth factor gene induction effectively promotes liver regeneration after hepatectomy in Solt-Farber rats. Hepatogastroenterology 2005; 52: 1393–7.
- 44 Casal M, Haskins M. Large animal models and gene therapy. Eur J Hum Genet 2006; 14: 266–72.
- 45 Dor Y, Djonov V, Abramovitch R, et al. Conditional switching of VEGF provides new insights into adult neovascularization and pro-angiogenic therapy. EMBO J 2002; 21: 1939–47.
- 46 Kay MA, Landen CN, Rothenberg SR, et al. In vivo hepatic gene therapy: complete albeit transient correction of factor IX deficiency in hemophilia B dogs. Proc Natl Acad Sci USA 1994; 91: 2353–7.
- 47 Gallo-Penn AM, Shirley PS, Andrews JL, et al. Systemic delivery of an adenoviral vector encoding canine factor VIII results in short-term phenotypic correction, inhibitor development, and biphasic liver toxicity in hemophilia A dogs. Blood 2001; 97: 107–13.
- 48 Ferrara N, Gerber H, LeCouter J. The biology of VEGF and its receptors. Nat Med 2003; 9: 669–76.
- 49 Moumen A, Ieraci A, Patané S, et al. Met signals hepatocyte survival by preventing Fas-triggered FLIP degradation in a PI3k-Akt-dependent manner. Hepatology 2007; 45: 1210–7.
- 50 Zhang HG, Xie J, Xu L, et al. Hepatic DR5 induces apoptosis and limits adenovirus gene therapy product expression in the liver. J Virol 2002; 76: 5692–700.
- 51 Okuyama T, Li XK, Funeshima N, et al. Fas-mediated apoptosis is involved in the elimination of gene-transduced hepatocytes with E1/E3-deleted adenoviral vectors. J Gastroenterol Hepatol 1998; 13 (Suppl.): S113–8.
- 52 Kosai K, Matsumoto K, Nagata S, et al. Abrogation of Fas-induced fulminant hepatic failure in mice by hepatocyte growth factor. Biochem Biophys Res Commun 1998; 244: 683–90.
- 53 Tanaka Y, Sohda T, Matsuo K, et al. Vascular endothelial growth factor reduces Fas-mediated acute liver injury in mice. J Gastroenterol Hepatol 2008; 23 (Part 2): e207–11.
- 54 Lusky M, Christ M, Rittner K, et al. In vitro and in vivo biology of recombinant adenovirus vectors with E1, E1/E2A, or E1/E4 deleted. J Virol 1998; 72: 2022–32.
- 55 Michou AI, Santoro L, Christ M, et al. Adenovirus-mediated gene therapy: influence of transgene, mouse strain and type of immune response on persistence of transgene expression. Gene Ther 1997; 4: 473–82.
- 56 Morral N, O'Neal W, Zhou H, et al. Immune response to reporter proteins and high viral dose limit duration of expression with adenoviral vectors: comparison of E2A wild-type and E2Adeleted vectors. Hum Gene Ther 1997; 8: 1275–86.
- 57 Tripathy SK, Black HB, Goldwasser E, Leiden JM. Immune responses to transgene-encoded proteins limit the stability of gene expression after injection of replication-defective adenovirus vectors. Nat Med 1996; 2: 545–50.
- 58 Lieber A, He CY, Meuse L, et al. The role of Kupffer cell activation and viral gene expression in early liver toxicity after infusion of recombinant adenovirus vectors. J Virol 1997; 71: 8798–807.
- 59 Wilson C, Kay MA. Immunomodulation to enhance gene therapy. Nat Med 1995; 1: 887–93.
- 60 Yang Y, Li Q, Ertl HC, Wilson JM. Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses. J Virol 1995; 69: 2004–15.
- 61 Peng Y, Trevejo J, Zhou J, et al. Inhibition of tumor necrosis factor alpha by an adenovirus-encoded soluble fusion protein extends transgene expression in the liver and lung. J Virol 1999; 73: 5098–109.
- 62 Raper SE, Chirmule N, Lee FS, et al. Fatal systemic inflammatory response syndrome in a ornithine transcarbamylase deficient patient following adenoviral gene transfer. Mol Genet Metab 2003; 80: 148–58.
- 63 Raper SE, Haskal ZJ, Ye X, et al. Selective gene transfer into the liver of non-human primates with E-1deleted, E2A-defective, or E-E4 deleted recombinant adenoviruses. Hum Gene Ther 1998; 9: 671–9.
- 64 Tao N, Gao G-P, Parr M, et al. Sequestration of adenoviral vector by Kupffer cells leads to a nonlinear dose response of transduction in liver. Mol Ther 2001; 3: 28–35.
- 65 Lin Y, Xie WF, Chen YX, et al. Treatment of experimental hepatic fibrosis by combinational delivery of urokinase-type plasminogen activator and hepatocyte growth factor genes. Liver Int 2005; 25: 796–807.
- 66 Kim WH, Matsumoto K, Bessho K, Nakamura T. Growth inhibition and apoptosis in liver myofibroblasts promoted by hepatocyte growth factor leads to resolution from liver cirrhosis. Am J Pathol 2005; 166: 1017–28.
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