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VEGF-D is an X-linked/AP-1 Regulated Putative Onco-angiogen in Human Glioblastoma Multiforme

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Abstract

Background

Glioblastoma multiforme (GBM) is a hypervascularized and locally infiltrating brain tumor of astroglial origin with a very poor prognosis. An X-linked c-fos oncogene-inducible mitogenic, morphogenic, and angiogenic factor, endothelial growth factor-D (VEGF-D), is the newest mammalian member of VEGF family. We analyzed VEGF-D in GBM because of its high angiogenic potential and its linkage to the X chromosome.

Materials and Methods

Nonmalignant brain and GBM tissue sections as well as GBM cell lines were analyzed by immunofluorescence for the expression of VEGF-D, factor VIII (endothelial cell marker), glial-fibrillary acidic protein (GFAP) (astrocytic cell lineage cytoplasmic marker), and several Fos family transcription factors, including c-Fos and Fra-1. The proteins were also detected by Western blots. The differences between genotypes of normal brain and GBM cells were examined by cDNA microarrays.

Results and Conclusions

GBM expressed ubiquitously VEGF-D, which colocalized with GFAP. Contrary to our expectations, low levels of c-Fos were detected in GBM cells. However, we identified another Fos family member, Fra-1, together with its transcriptional activation partner, c-Jun, as being stably up-regulated in GBM cells. Furthermore, we demonstrated that a fra-1 transgene induced VEGF-D expression in cultured cells and GBM cell stimulation evoked a sustained increase in both Fra-1 and VEGF-D levels. This study reveals that an up-regulation of AP-1 factors may be a hallmark of GBM. Because VEGF-D activates VEGF receptor 2 and 3, receptors important for tumor angiogenesis, it may represent an X-linked/AP-1-regulated onco-angiogen in human GBM. The VEGF-D system and AP-1 activity appear to be very attractive targets for new molecular diagnostics and rational molecular anti-cancer therapies.

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References

  1. Salcman M. (1993). Concepts in neurosurgery: neurobiology of brain tumors. Baltimore, MD: Williams & Wilkins.

    Google Scholar 

  2. Long DM. (1970). Capillary ultrastructure and the blood-brain barrier in human malignant brain tumors. J. Neurosurg. 32: 127–144.

    Article  CAS  PubMed  Google Scholar 

  3. Goldman CK, Kim J, Wong W-L, King V, Brock T, Gillespie GY. (1993). Epidermal growth factor stimulates vascular endothelial growth factor production by human malignant glioma cells: a model of glioblastoma multiforme pathophysiology. Mol. Biol. Cell 4: 121–133.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Kerbel RS, Viloria-Petit A, Okada F, Rak J. (1998). Establishing a link between oncogenes and tumor angiogenesis. Mol. Med. 4: 286–295.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Orlandini M, Larconcini L, Ferruzzi R, Oliviero S. (1996). Identification of a c-fos-induced gene that is related to the platelet-derived growth factor/vascular endothelial growth factor family. Proc. Natl. Acad. Sci. U.S.A. 93: 11675–11680.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Achen MG, Jeltsch M, Kukk E, et al. (1998). Vascular endothelial growth factor D (VEGF-D) is a ligand for the tyrosine kinases VEGF receptor 2 (Flk1) and VEGF receptor 3 (Flt4). Proc. Natl. Acad. Sci. U.S.A. 95: 548–553.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Marconcini L, Marchio S, Morbidelli L, et al. (1999). c-fos-induced growth factor/vascular endothelial growth factor D induces angiogenesis in vivo and in vitro. Proc. Natl. Acad. Sci. U.S.A. 96: 9671–9676.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Debinski W, Gibo D, Hulet SW, Connor JR, Gillespie GY. (1999). Receptor for interleukin 13 is a marker and therapeutic target for human high-grade glioma Clin. Cancer Res. 5: 985–990.

    PubMed  CAS  Google Scholar 

  9. Debinski W, Gibo D, Obiri N, Kealiher A, Puri R. (1998). Novel anti-brain tumor cytotoxins specific for cancer cells. Nature Biotech. 16: 449–453.

    Article  CAS  Google Scholar 

  10. Debinski W, Gibo DM. (2000). Molecular expression analysis of restrictive receptor for interleukin 13, a brain tumor-associated cancer/testis antigen. Mol. Med. 6: 440–449.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Debinski W, Slagle B, Gibo DM, Powers SK, Gillespie GY. (2000). Expression of a restrictive receptor for interleukin 13 is associated with glial transformation. J. Neurooncol. 48: 103–111.

    Article  CAS  PubMed  Google Scholar 

  12. Old LJ, Chen TC. (1998). New paths in human cancer serology. J. Exp. Med. 187: 1163–1167.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Mintz A, Debinski W. (2000). Cancer genetics/epigenetics and the X chromosome: Possible new links for malignant glioma pathogenesis and immune-based therapy. Crit. Rev. Oncogen. 11: 77–95.

    Article  CAS  Google Scholar 

  14. Jenkins NA, Woolatt E, Crawford J, et al. (1997). Mapping of the gene for vascular endothelial growth factor-D in mouse and man to the X chromosome. Chromosome Res. 5: 502–505.

    PubMed  CAS  Google Scholar 

  15. Achen MG, Roufail S, Domagala T, et al. (2000). Monoclonal antibodies to vascular endothelial growth factor-D block its interaction with both VEGF receptor-2 and VEGF receptor-3. Eur. J. Biochem. 267: 2505–2515.

    Article  CAS  PubMed  Google Scholar 

  16. Chomczynski P, Sacchi N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162: 156–159.

    Article  CAS  PubMed  Google Scholar 

  17. Rocchigiani M, Lestingi M, Luddi A, et al. (1998). Human FIGF: cloning, gene structure, and mapping to chromosome Xp22.1 between the PIGA and the GRPR genes. Genomics 47: 207–216.

    Article  CAS  PubMed  Google Scholar 

  18. McKeever PE. (1998). Insights about brain tumors gained through immunohistochemistry and in situ hybridization of nuclear and phenotypic markers. J. Histochem. Cytochem. 46: 585–594.

    Article  CAS  PubMed  Google Scholar 

  19. Stacker SA, Stenvers K, Caesar C, et al. (1999) Biosynthesis of vascular endothelial growth factor-D involves proteolytic processing which generates non-covalent homodimers. J. Biol. Chem. 274: 32127–32136.

    Article  CAS  PubMed  Google Scholar 

  20. Halfter H, Kremerskothen J, Weber J, et al. (1998). Growth inhibition of newly established human glioma cell lines by leukemia inhibitory factor. J. Neurooncol. 39: 1–18.

    Article  CAS  PubMed  Google Scholar 

  21. Bergers G, Graninger P, Braselmann S, Wrighton C, Busslinger M. (1995). Transcriptional activation of the fra-1 gene by AP-1 is mediated by regulatory sequences in the first intron. Mol. Cell. Biol. 15: 3748–3758.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Kustikova O, Kramerov D, Grigorian M, et al. (1998). Fra-1 induces morphological transformation and increases in vitro invasiveness and motility of epithelioid adenocarcinoma cells. Mol. Cell. Biol. 18: 7095–7105.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Folkman J. (1995). Angiogenesis in cancer, vascular, rheumatoid and other diseases. Nat. Med. 1: 27–31.

    Article  CAS  PubMed  Google Scholar 

  24. Brem S, Cotran R, Folkman J. (1972). Tumor angiogenesis: a quantitative method for histologic grading. J. Natl. Cancer Inst. 48: 347–356.

    PubMed  CAS  Google Scholar 

  25. Tulchinsky E. (2000). Fos family members: regulation, structure, and role in oncogenic transformation. Histol. Histopathol. 15: 921–928.

    PubMed  CAS  Google Scholar 

  26. Lilja A, Stanisic M, Salford L. (2000). Abnormal expression of OSM, CNTF and LIF in human brain tumors. Neuro-Oncology 2: S22.

    Article  Google Scholar 

  27. Zagzag D, Miller DC, Sato Y, Rifkin DB, Burstein DE. (1990). Immunohistochemical localization of basic fibroblast growth factor in astrocytomas. Cancer Res. 50: 7393–7398.

    PubMed  CAS  Google Scholar 

  28. Botelho FM, Edwards DR, Richards CD. (1998). Oncostatin M stimulates c-Fos to bind a transcriptionally responsive AP-1 element within the tissue inhibitor of metalloproteinase-1 promoter. J. Biol. Chem. 273: 5211–5218.

    Article  CAS  PubMed  Google Scholar 

  29. Mohan R, Sivak J, Ashton P, et al. (2000). Curcuminoids inhibit the angiogenic response stimulated by fibroblast growth factor-2, including expression of matrix metalloproteinase gelatinase B. J. Biol. Chem. 275: 10405–10412.

    Article  CAS  PubMed  Google Scholar 

  30. Mechta F, Lallemand D, Pfarr CM, Yaniv M. (1997). Transformation by ras modifies AP1 composition and activity. Oncogene 14: 837–847.

    Article  CAS  PubMed  Google Scholar 

  31. Nakano A, Tani E, Miyazaki K, Yamamoto Y, Furuyama J-I. (1995). Matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gliomas. J. Neurosurg. 83: 298–307.

    Article  CAS  PubMed  Google Scholar 

  32. Schreiber M, Wand Z-Q, Jochum W, Fetka I, Elliott C, Wagner EF. (2000). Placental vascularisation requires the AP-1 component Fra-1. Development 127: 4937–4948.

    PubMed  CAS  Google Scholar 

  33. Phillips BW, Sharma R, Leco PA, Edwards DR. (1999). A sequence-selective single-strand DNA-binding protein regulates basal transcription of the murine tissue inhibitor of metalloproteinases-1 (Timp-1) gene. J. Biol. Chem. 274: 22197–22207.

    Article  CAS  PubMed  Google Scholar 

  34. Murakami M, Motoyasu U, Iba H. (1999). Fra-2-positive auto-regulatory loop triggered by mitogen-activated protein kinase (MAPK) and Fra-2 phosphorylation sites by MAPK. Cell Growth Differ. 10: 333–342.

    PubMed  CAS  Google Scholar 

  35. Ransone LJ, Verma IM. (1990). Nuclear proto-oncogenes fos and jun. Annu. Rev. Cell Biol. 6: 539–557.

    Article  CAS  PubMed  Google Scholar 

  36. Kovary K, Bravo R. (1992). Existence of different Fos/Jun complexes during the G0-to-G1 transition and during exponential growth in mouse fibroblasts: differential role of Fos proteins. Mol. Cell. Biol. 12: 5015–5023.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Gruda MC, Kovary K, Metz R, Bravo R. (1994). Regulation of Fra-1 and Fra-2 phosphorylation differs during the cell cycle of fibroblasts and phosphorylation in vitro by MAP kinase affects DNA binding activity. Oncogene 9: 2537–2547.

    PubMed  CAS  Google Scholar 

  38. Fleischmann A, Hafezi F, Elliott C, Reme CE, Ruther U, Wagner EF. (2000). Fra-1 replaces c-Fos-dependent functions in mice. Genes Dev. 14: 2695–2700.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Mishima K, Mazar AP, Gown A, et al. (2000). A peptide derived from the non-receptor binding region of urokinase plasminogen activator inhibits glioblastoma growth and angiogenesis in vivo in combination with cisplatin. Proc. Natl. Acad. Sci. U.S.A. 97: 8484–8489.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Millauer B, Shawver LK, Plate KH, Risau W, Ullrich A. (1994). Glioblastoma growth inhibited in vivo by a dominant-negative Flk-1 mutant. Nature 367: 576–579.

    Article  CAS  PubMed  Google Scholar 

  41. Takano S, Yoshii Y, Kondo S, et al. (1996). Concentrations of vascular endothelial growth factor in the serum and tumor tissue of brain tumor patients. Cancer Res. 56: 2185–2190.

    PubMed  CAS  Google Scholar 

  42. Strawn LM, McMahon G, App H, et al. (1996) Flk-1 as a target for tumor growth inhibition. Cancer Res. 56: 3540–3545.

    PubMed  CAS  Google Scholar 

  43. Skobe M, Rockwell P, Goldstein N, Vosseler S, Fusenig NE. (1997). Halting angiogenesis suppresses carcinoma cell invasion. Nat. Med. 3: 1222–1227.

    Article  CAS  PubMed  Google Scholar 

  44. Prewett M, Huber J, Li Y, et al. (1999) Anti-vascular endothelial growth factor receptor (Fetal Liver Kinase1) monoclonal antibody inhibits tumor angiogenesis and growth of several mouse and human tumors. Cancer Res. 59: 5209–5218.

    PubMed  CAS  Google Scholar 

  45. Rubenstein JL, Kim J, Ozawa T, et al. (2000). Anti-VEGF antibody treatment of glioblastoma prolongs survival but results in increased vascular cooption. Neoplasia 2: 306–314.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Okada F, Rak J, St. Croix B, et al. (1998). Impact of oncogenes in tumor angiogenesis: mutant K-ras up-regulation of vascular endothelial growth factor/vascular permeability factor is necessary, but not sufficient for tumorigenicity of human colorectal carcinoma cells. Proc. Natl. Acad. Sci. U.S.A. 95: 3609–3614.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Stacker SA, Caesar C, Baldwin ME, et al. (2001). VEGF-D promotes the metastatic spread of tumor cells by the lymphatics. Nat. Med. 7: 168–175.

    Article  CAS  Google Scholar 

  48. Battista S, de Nigris F, Fedele M, et al. (1998). Increase in AP-1 activity is a general event in thyroid cell transformation in vitro and in vivo. Oncogene 17: 377–385.

    Article  CAS  PubMed  Google Scholar 

  49. Zoumpourlis V, Papassava P, Lindardopoulos S, Gillespie D, Balmain A, Pintzas A. (2000). High levels of c-Jun, Fra-1, Fra-2 and ATF-2 proteins correlate with malignant phenotypes in the multistage mouse skin carcinogenesis model. Oncogene 19: 4011–4021.

    Article  CAS  PubMed  Google Scholar 

  50. Bancroft CC, Chen Z, Dong G, et al. (2001). Coexpression of proangiogenic factors IL-8 and VEGF by human head and neck squamous cell carcinoma involves coactivation by MEK-MAPK and IKK-NF-kB signal pathways. Clin. Cancer Res. 7: 435–442.

    PubMed  CAS  Google Scholar 

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Acknowledgments

The American Cancer Society (W.D.) supported this work. We thank all neurosurgeons in the Section of Neurosurgery at Hershey who helped to secure tumor samples, especially Dr. Stephen K. Powers. We thank Dr. Stephen B. Baylin for comments on the manuscript. M.G.A. and S.A.S. were supported by the National Health and Medical Research Council of Australia and the Anti-Cancer Council of Victoria. E.T. was supported by the Danish Cancer Society.

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Authors and Affiliations

  1. Division of Neurosurgery/H110, Pennsylvania State University College of Medicine, Hershey, Pennsylvania, USA

    Waldemar Debinski MD, PhD (Director of Tumor Research), Becky Slagle-Webb & Denise M. Gibo

  2. Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Melbourne, Victoria, Australia

    Marc G. Achen & Steven A. Stacker

  3. Department of Molecular Cancer Biology, Danish Cancer Society, Copenhagen, Denmark

    Eugene Tulchinsky

  4. Division of Neurosurgery, Department of Surgery, The University of Alabama at Birmingham, School of Medicine, Birmingham, Alabama, USA

    G. Yancey Gillespie

  5. Section of Neurosurgery/H110, Department of Surgery, Pennsylvania State University College of Medicine, 500 University Drive, Hershey, Pennsylvania, 17033-0850, USA

    Waldemar Debinski MD, PhD (Director of Tumor Research)

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  1. Waldemar Debinski MD, PhD
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Correspondence to Waldemar Debinski MD, PhD.

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Debinski, W., Slagle-Webb, B., Achen, M.G. et al. VEGF-D is an X-linked/AP-1 Regulated Putative Onco-angiogen in Human Glioblastoma Multiforme. Mol Med 7, 598–608 (2001). https://doi.org/10.1007/BF03401866

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