Differential regulation of tumor angiogenesis by distinct ErbB homo- and heterodimers.

Interactions between cancer cells and their microenvironment are critical for the development and progression of solid tumors. This study is the first to examine the role of all members of the ErbB tyrosine kinase receptors (epidermal growth factor receptor [EGFR], ErbB-2, ErbB-3, or ErbB-4), expressed singly or as paired receptor combinations, in the regulation of angiogenesis both in vitro and in vivo. Comparison of all receptor combinations reveals that EGFR/ErbB-2 and ErbB-2/ErbB-3 heterodimers are the most potent inducers of vascular endothelial growth factor (VEGF) mRNA expression compared with EGFR/ErbB-3, EGFR/ErbB-4, ErbB-2/ErbB-4, and ErbB-3/ErbB-4. Immunohistochemistry of tumor xenografts overexpressing these heterodimers shows increased VEGF expression and remarkably enhanced vascularity. Enhanced VEGF expression is associated with increased VEGF transcription. Deletional analysis reveals that ErbB-mediated transcriptional up-regulation of VEGF involves a hypoxia-inducible factor 1-independent responsive region located between nucleotides -88 to -66 of the VEGF promoter. Mutational analysis reveals that the Sp-1 and AP-2 transcription factor binding elements within this region are required for up-regulation of VEGF by heregulin beta1 and that this up-regulation is dependent on the activity of extracellular signal-related protein kinases. These results emphasize the biological implications of cell signaling diversity among members of the ErbB receptor family in regulation of the tumor microenvironment.

[1]  Y. Yarden,et al.  Untangling the ErbB signalling network , 2001, Nature Reviews Molecular Cell Biology.

[2]  N. Ferrara,et al.  The biology of vascular endothelial growth factor. , 1997, Endocrine reviews.

[3]  M. Makuuchi,et al.  Hypoxia response element of the human vascular endothelial growth factor gene mediates transcriptional regulation by nitric oxide: control of hypoxia-inducible factor-1 activity by nitric oxide. , 2000, Blood.

[4]  L. Cantley,et al.  Insect cell-expressed p180erbB3 possesses an impaired tyrosine kinase activity. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[5]  A. Thomason,et al.  Transformation of NIH 3T3 Cells by HER3 or HER4 Receptors Requires the Presence of HER1 or HER2 (*) , 1996, The Journal of Biological Chemistry.

[6]  Y. Yarden,et al.  Neu differentiation factor activation of ErbB-3 and ErbB-4 is cell specific and displays a differential requirement for ErbB-2 , 1995, Molecular and cellular biology.

[7]  N. Normanno,et al.  Epidermal growth factor-related peptides and their receptors in human malignancies. , 1995, Critical reviews in oncology/hematology.

[8]  T. van Raaij,et al.  The cellular response to neuregulins is governed by complex interactions of the erbB receptor family , 1995, Molecular and cellular biology.

[9]  M. Kraus,et al.  Oncogenic potential of erbB-2 in human mammary epithelial cells. , 1991, Oncogene.

[10]  C. Van Waes,et al.  Expression of proangiogenic chemokine Gro 1 in low and high metastatic variants of Pam murine squamous cell carcinoma is differentially regulated by IL‐1α, EGF and TGF‐β1 through NF‐κB dependent and independent mechanisms , 2001 .

[11]  K. Alitalo,et al.  Structure, expression and receptor-binding properties of novel vascular endothelial growth factors. , 1999, Current topics in microbiology and immunology.

[12]  F. Leenders,et al.  Mammary gland specific hEGF receptor transgene expression induces neoplasia and inhibits differentiation , 2000, Oncogene.

[13]  Goldman,et al.  Epidermal growth factor stimulates vascular endothelial growth factor production by human malignant glioma cells: a model of glioblastoma multiforme pathophysiology. , 1993, Molecular biology of the cell.

[14]  J. Folkman.  What is the evidence that tumors are angiogenesis dependent? , 1990, Journal of the National Cancer Institute.

[15]  Jessica Lo,et al.  HIF‐1α is required for solid tumor formation and embryonic vascularization , 1998 .

[16]  P. Carmeliet,et al.  Role of HIF-1α in hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis , 1998, Nature.

[17]  L. Cantley,et al.  A neu acquaintance for ErbB3 and ErbB4: A role for receptor heterodimerization in growth signaling , 1994, Cell.

[18]  L. Claesson‐Welsh,et al.  FGF and VEGF function in angiogenesis: signalling pathways, biological responses and therapeutic inhibition. , 2001, Trends in pharmacological sciences.

[19]  M. Greene,et al.  Intermolecular association of the p185 neu protein and EGF receptor modulates EGF receptor function , 1990, Cell.

[20]  D. Taverna,et al.  NDF/heregulin activates MAP kinase and p70/p85 S6 kinase during proliferation or differentiation of mammary epithelial cells. , 1995, Oncogene.

[21]  Y. Yarden,et al.  Diversification of Neu differentiation factor and epidermal growth factor signaling by combinatorial receptor interactions. , 1996, The EMBO journal.

[22]  B. Zetter,et al.  Angiogenesis and tumor metastasis. , 1998, Annual review of medicine.

[23]  Y. Yarden,et al.  A hierarchical network of interreceptor interactions determines signal transduction by Neu differentiation factor/neuregulin and epidermal growth factor , 1996, Molecular and cellular biology.

[24]  V. Brown,et al.  Synergistic interaction of p185c-neu and the EGF receptor leads to transformation of rodent fibroblasts , 1989, Cell.

[25]  R. Kerbel,et al.  Mutant ras oncogenes upregulate VEGF/VPF expression: implications for induction and inhibition of tumor angiogenesis. , 1995, Cancer research.

[26]  N. Hynes,et al.  Single-chain antibody-mediated intracellular retention of ErbB-2 impairs Neu differentiation factor and epidermal growth factor signaling , 1995, Molecular and cellular biology.

[27]  N. Hynes,et al.  Intracellular expression of a single-chain antibody directed to the EGFR leads to growth inhibition of tumor cells. , 1996, Oncogene.

[28]  D. Mukhopadhyay,et al.  Hypoxic induction of human vascular endothelial growth factor expression through c-Src activation , 1995, Nature.

[29]  K. Kuroi,et al.  Prognostic significance of co-expression of c-erbB-2 oncoprotein and epidermal growth factor receptor in breast cancer patients. , 1992, American journal of surgery.

[30]  N. Andrews,et al.  A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. , 1991, Nucleic acids research.

[31]  H. Phillips,et al.  c‐erbb growth‐factor‐receptor proteins in ovarian tumours , 1995, International journal of cancer.

[32]  Y. Yarden,et al.  ErbB‐2 is a common auxiliary subunit of NDF and EGF receptors: implications for breast cancer. , 1996, The EMBO journal.

[33]  G. Semenza,et al.  Modulation of hypoxia-inducible factor 1alpha expression by the epidermal growth factor/phosphatidylinositol 3-kinase/PTEN/AKT/FRAP pathway in human prostate cancer cells: implications for tumor angiogenesis and therapeutics. , 2000, Cancer research.

[34]  H. P. Fell,et al.  The Relationship between Human Epidermal Growth-like Factor Receptor Expression and Cellular Transformation in NIH3T3 Cells* , 1996, The Journal of Biological Chemistry.

[35]  B. Slagle,et al.  Transcriptional Regulation of the TFIIH Transcription Repair Components XPB and XPD by the Hepatitis B Virus x Protein in Liver Cells and Transgenic Liver Tissue* , 2001, The Journal of Biological Chemistry.

[36]  M. Kraus,et al.  Cooperative signaling of ErbB3 and ErbB2 in neoplastic transformation and human mammary carcinomas. , 1995, Oncogene.

[37]  N. Goldstein,et al.  Neutralizing antibodies against epidermal growth factor and ErbB-2/neu receptor tyrosine kinases down-regulate vascular endothelial growth factor production by tumor cells in vitro and in vivo: angiogenic implications for signal transduction therapy of solid tumors. , 1997, The American journal of pathology.

[38]  M. Krasnow,et al.  The Hypoxic Response: Huffing and HIFing , 1997, Cell.

[39]  C R King,et al.  erbB-2 is a potent oncogene when overexpressed in NIH/3T3 cells. , 1987, Science.

[40]  G. Semenza,et al.  Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1 , 1996, Molecular and cellular biology.

[41]  Neu Differentiation Factor Stimulates Phosphorylation and Activation of the Sp1 Transcription Factor , 1999, Molecular and Cellular Biology.

[42]  R. Cardiff,et al.  Elevated expression of activated forms of Neu/ErbB‐2 and ErbB‐3 are involved in the induction of mammary tumors in transgenic mice: implications for human breast cancer , 1999, The EMBO journal.

[43]  A. Ullrich,et al.  Heregulin‐dependent regulation of HER2/neu oncogenic signaling by heterodimerization with HER3. , 1995, The EMBO journal.

[44]  N. Ferrara.  Molecular and biological properties of vascular endothelial growth factor , 1999, Journal of Molecular Medicine.

[45]  Monilola A. Olayioye,et al.  The ErbB signaling network: receptor heterodimerization in development and cancer , 2000, The EMBO journal.

[46]  N. Hynes,et al.  The biology of erbB-2/neu/HER-2 and its role in cancer. , 1994, Biochimica et biophysica acta.

[47]  D. Marmé,et al.  Both v-Ha-Ras and v-Raf Stimulate Expression of the Vascular Endothelial Growth Factor in NIH 3T3 Cells (*) , 1995, The Journal of Biological Chemistry.

[48]  J. Caro,et al.  Hypoxia regulatory elements of the human vascular endothelial growth factor gene. , 1994, Cellular & molecular biology research.

[49]  M. Alaoui-Jamali,et al.  Dual effect of erbB-2 depletion on the regulation of DNA repair and cell cycle mechanisms in non-small cell lung cancer cells , 1998, Oncogene.

[50]  M. Sliwkowski,et al.  Coexpression of erbB2 and erbB3 proteins reconstitutes a high affinity receptor for heregulin. , 1994, The Journal of biological chemistry.

[51]  Y. Yarden,et al.  An Immunological Approach Reveals Biological Differences between the Two NDF/Heregulin Receptors, ErbB-3 and ErbB-4 (*) , 1996, The Journal of Biological Chemistry.

[52]  A. Ullrich,et al.  Overexpression of the human EGF receptor confers an EGF-dependent transformed phenotype to NIH 3T3 cells , 1987, Cell.

[53]  Y. Yarden,et al.  The ErbB signaling network in embryogenesis and oncogenesis: signal diversification through combinatorial ligand‐receptor interactions , 1997, FEBS letters.

[54]  D. Stern,et al.  Specificity within the EGF family/ErbB receptor family signaling network , 1998, BioEssays : news and reviews in molecular, cellular and developmental biology.

[55]  S. Mader,et al.  Heregulin selectively upregulates vascular endothelial growth factor secretion in cancer cells and stimulates angiogenesis , 2000, Oncogene.

[56]  J. Pouysségur,et al.  p42/p44 MAP Kinase Module Plays a Key Role in the Transcriptional Regulation of the Vascular Endothelial Growth Factor Gene in Fibroblasts* , 1998, The Journal of Biological Chemistry.

[57]  Y. Chiew,et al.  Expression of c‐erbB receptors, heregulin and oestrogen receptor in human breast cell lines , 2000, International journal of cancer.

[58]  J. Nalbantoglu,et al.  Vesicular stomatitis virus G pseudotyped retrovector mediates effective in vivo suicide gene delivery in experimental brain cancer. , 1999, Cancer research.

[59]  G. Semenza,et al.  HER2 (neu) Signaling Increases the Rate of Hypoxia-Inducible Factor 1α (HIF-1α) Synthesis: Novel Mechanism for HIF-1-Mediated Vascular Endothelial Growth Factor Expression , 2001, Molecular and Cellular Biology.

[60]  M. Sliwkowski,et al.  Growth regulation of human breast and ovarian tumor cells by heregulin: Evidence for the requirement of ErbB2 as a critical component in mediating heregulin responsiveness. , 1996, Cancer research.

[61]  H. Modjtahedi,et al.  Vascular endothelial growth factor family members are differentially regulated by c-erbB signaling in head and neck squamous carcinoma cells , 2004, Clinical & Experimental Metastasis.

[62]  N. Hynes,et al.  ErbB‐2, the preferred heterodimerization partner of all ErbB receptors, is a mediator of lateral signaling , 1997, The EMBO journal.

[63]  E. De Clercq,et al.  Angiogenesis: regulators and clinical applications. , 2001, Biochemical pharmacology.