Fully human monoclonal antibodies to hepatocyte growth factor with therapeutic potential against hepatocyte growth factor/c-Met-dependent human tumors.

c-Met is a well-characterized receptor tyrosine kinase for hepatocyte growth factor (HGF). Compelling evidence from studies in human tumors and both cellular and animal tumor models indicates that signaling through the HGF/c-Met pathway mediates a plethora of normal cellular activities, including proliferation, survival, migration, and invasion, that are at the root of cancer cell dysregulation, tumorigenesis, and tumor metastasis. Inhibiting HGF-mediated signaling may provide a novel therapeutic approach for treating patients with a broad spectrum of human tumors. Toward this goal, we generated and characterized five different fully human monoclonal antibodies that bound to and neutralized human HGF. Antibodies with subnanomolar affinities for HGF blocked binding of human HGF to c-Met and inhibited HGF-mediated c-Met phosphorylation, cell proliferation, survival, and invasion. Using a series of human-mouse chimeric HGF proteins, we showed that the neutralizing antibodies bind to a unique epitope in the beta-chain of human HGF. Importantly, these antibodies inhibited HGF-dependent autocrine-driven tumor growth and caused significant regression of established U-87 MG tumor xenografts. Treatment with anti-HGF antibody rapidly inhibited tumor cell proliferation and significantly increased the proportion of apoptotic U-87 MG tumor cells in vivo. These results suggest that an antibody to an epitope in the beta-chain of HGF has potential as a novel therapeutic agent for treating patients with HGF-dependent tumors.

[1]  D. Bottaro,et al.  Hereditary papillary renal carcinoma type I. , 2004, Current molecular medicine.

[2]  L. Trusolino,et al.  Scatter-factor and semaphorin receptors: cell signalling for invasive growth , 2002, Nature Reviews Cancer.

[3]  R. Ondarza,et al.  Hepatocyte growth factor is associated with poor prognosis of malignant gliomas and is a predictor for recurrence of meningioma , 2002, Cancer.

[4]  P. Godowski,et al.  Generation and characterization of a competitive antagonist of human hepatocyte growth factor, HGF/NK1. , 1993, The Journal of biological chemistry.

[5]  Toshikazu Nakamura,et al.  NK4 (HGF‐antagonist/angiogenesis inhibitor) in cancer biology and therapeutics , 2003, Cancer science.

[6]  K. Matsumoto,et al.  Elevation of serum hepatocyte growth factor concentration in patients with gastric cancer is mediated by production from tumor tissue. , 2000, Anticancer research.

[7]  A. Bardelli,et al.  Mutant Met-mediated transformation is ligand-dependent and can be inhibited by HGF antagonists , 1999, Oncogene.

[8]  J. Rubin,et al.  A broad-spectrum human lung fibroblast-derived mitogen is a variant of hepatocyte growth factor. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[9]  A. Waage,et al.  Hepatocyte growth factor in myeloma patients treated with high‐dose chemotherapy , 2002, British journal of haematology.

[10]  E. Rosen,et al.  Regulation of angiogenesis by scatter factor. , 1997, EXS.

[11]  E. Hudson,et al.  Met and hepatocyte growth factor/scatter factor expression in human gliomas. , 1997, Cancer research.

[12]  Thomas Hartmann,et al.  Suppression of angiogenesis and tumor growth by selective inhibition of angiopoietin-2. , 2004, Cancer cell.

[13]  W. Birchmeier,et al.  Met, metastasis, motility and more , 2003, Nature Reviews Molecular Cell Biology.

[14]  B. Elliott,et al.  Coexpression of hepatocyte growth factor and receptor (Met) in human breast carcinoma. , 1996, The American journal of pathology.

[15]  A. Bell,et al.  The five amino acid-deleted isoform of hepatocyte growth factor promotes carcinogenesis in transgenic mice , 1999, Oncogene.

[16]  J. Rubin,et al.  Hepatocyte growth factor/scatter factor overexpression induces growth, abnormal development, and tumor formation in transgenic mouse livers. , 1996, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[17]  P. Godowski,et al.  Structure‐function analysis of hepatocyte growth factor: identification of variants that lack mitogenic activity yet retain high affinity receptor binding. , 1992, The EMBO journal.

[18]  G. Woude,et al.  HGF/SF-Met signaling in tumor progression , 2005, Cell Research.

[19]  M. Fiscella,et al.  The mutationally activated Met receptor mediates motility and metastasis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Christian Wiesmann,et al.  Crystal structure of the HGF β‐chain in complex with the Sema domain of the Met receptor , 2004, The EMBO journal.

[21]  L. Presta,et al.  Mutational analysis and molecular modeling of the N-terminal kringle-containing domain of hepatocyte growth factor identifies amino acid side chains important for interaction with the c-Met receptor. , 1994, Protein engineering.

[22]  J. Rubin,et al.  Identification of a competitive HGF antagonist encoded by an alternative transcript. , 1991, Science.

[23]  G. V. Vande Woude,et al.  Neutralizing monoclonal antibodies to hepatocyte growth factor/scatter factor (HGF/SF) display antitumor activity in animal models , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[24]  W. K. Alfred Yung,et al.  Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers , 1997, Nature Genetics.

[25]  G. Woude,et al.  Enhanced growth of human met-expressing xenografts in a new strain of immunocompromised mice transgenic for human hepatocyte growth factor/scatter factor , 2005, Oncogene.

[26]  J. Beckstead,et al.  A simple technique for preservation of fixation-sensitive antigens in paraffin-embedded tissues. , 1994, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[27]  D. Kirchhofer,et al.  Structural and Functional Basis of the Serine Protease-like Hepatocyte Growth Factor β-Chain in Met Binding and Signaling* , 2004, Journal of Biological Chemistry.

[28]  J. Thiery.  Epithelial–mesenchymal transitions in tumour progression , 2002, Nature Reviews Cancer.

[29]  Larry L. Green,et al.  Functional transplant of megabase human immunoglobulin loci recapitulates human antibody response in mice , 1997, Nature Genetics.

[30]  T. Blundell,et al.  Molecular evolution and domain structure of plasminogen‐related growth factors (HGF/SF and HGF1/MSP) , 1994, Protein science : a publication of the Protein Society.

[31]  K. Miyazaki,et al.  Expression of the hepatocyte growth factor/c‐Met pathway is increased at the cancer front in breast carcinoma , 2001, Pathology international.

[32]  R. Sharp,et al.  Diverse tumorigenesis associated with aberrant development in mice overexpressing hepatocyte growth factor/scatter factor. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[33]  L. Naldini,et al.  Hepatocyte growth factor is a potent angiogenic factor which stimulates endothelial cell motility and growth , 1992, The Journal of cell biology.

[34]  M. Fiscella,et al.  Activating Met mutations produce unique tumor profiles in mice with selective duplication of the mutant allele. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[35]  Y. Cho,et al.  Significant Correlation between Serum Level of Hepatocyte Growth Factor and Progression of Gastric Carcinoma , 1999, World Journal of Surgery.

[36]  G. Merlino,et al.  NK1, a Natural Splice Variant of Hepatocyte Growth Factor/Scatter Factor, Is a Partial Agonist In Vivo , 1998, Molecular and Cellular Biology.

[37]  N. Ferrara,et al.  The biology of VEGF and its receptors , 2003, Nature Medicine.