EphA3 maintains tumorigenicity and is a therapeutic target in glioblastoma multiforme.

Significant endeavor has been applied to identify functional therapeutic targets in glioblastoma (GBM) to halt the growth of this aggressive cancer. We show that the receptor tyrosine kinase EphA3 is frequently overexpressed in GBM and, in particular, in the most aggressive mesenchymal subtype. Importantly, EphA3 is highly expressed on the tumor-initiating cell population in glioma and appears critically involved in maintaining tumor cells in a less differentiated state by modulating mitogen-activated protein kinase signaling. EphA3 knockdown or depletion of EphA3-positive tumor cells reduced tumorigenic potential to a degree comparable to treatment with a therapeutic radiolabelled EphA3-specific monoclonal antibody. These results identify EphA3 as a functional, targetable receptor in GBM.

[1]  E. Fokas,et al.  Increased expression of EphA2 correlates with adverse outcome in primary and recurrent glioblastoma multiforme patients. , 2008, Oncology reports.

[2]  D. Wilkinson,et al.  Eph receptors and ephrins: regulators of guidance and assembly. , 2000, International review of cytology.

[3]  S. Weiss,et al.  Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. , 1992, Science.

[4]  Elena B. Pasquale,et al.  Eph receptors and ephrins in cancer: bidirectional signalling and beyond , 2010, Nature Reviews Cancer.

[5]  D. Xie,et al.  EphrinA5 acts as a tumor suppressor in glioma by negative regulation of epidermal growth factor receptor , 2009, Oncogene.

[6]  Michael F. Clarke,et al.  Applying the principles of stem-cell biology to cancer , 2003, Nature Reviews Cancer.

[7]  E. Fokas,et al.  Increased expression of EphA7 correlates with adverse outcome in primary and recurrent glioblastoma multiforme patients , 2008, BMC Cancer.

[8]  M. Lackmann,et al.  Ephrin-A5 induces rounding, blebbing and de-adhesion of EphA3-expressing 293T and melanoma cells by CrkII and Rho-mediated signalling. , 2002, Journal of cell science.

[9]  K. Hoang-Xuan,et al.  Primary brain tumours in adults , 2003, The Lancet.

[10]  Mark W. Dewhirst,et al.  Glioma stem cells promote radioresistance by preferential activation of the DNA damage response , 2006, Nature.

[11]  R. McLendon,et al.  Integrin alpha 6 regulates glioblastoma stem cells. , 2010, Cell stem cell.

[12]  P. Coulie,et al.  Identification of a tumor-specific shared antigen derived from an Eph receptor and presented to CD4 T cells on HLA class II molecules. , 2000, Cancer research.

[13]  J. Dairiki.  Radioactive Decay Data Tables: A Handbook of Decay Data for Application to Radiation Dosimetry and Radiological Assessments , 1982 .

[14]  R. Klein,et al.  Eph receptors and ephrins: effectors of morphogenesis. , 1999, Development.

[15]  T. Yamashita,et al.  EphA Receptors Direct the Differentiation of Mammalian Neural Precursor Cells through a Mitogen-activated Protein Kinase-dependent Pathway* , 2004, Journal of Biological Chemistry.

[16]  Jonas Frisén,et al.  EphB Receptors Coordinate Migration and Proliferation in the Intestinal Stem Cell Niche , 2006, Cell.

[17]  A. Scott,et al.  Concurrent binding of anti-EphA3 antibody and ephrin-A5 amplifies EphA3 signaling and downstream responses: potential as EphA3-specific tumor-targeting reagents. , 2005, Cancer research.

[18]  Sieger Leenstra,et al.  Novel somatic and germline mutations in cancer candidate genes in glioblastoma, melanoma, and pancreatic carcinoma. , 2007, Cancer research.

[19]  J G Flanagan,et al.  The ephrins and Eph receptors in neural development. , 1998, Annual review of neuroscience.

[20]  Elena B Pasquale,et al.  Cancer somatic mutations disrupt functions of the EphA3 receptor tyrosine kinase through multiple mechanisms. , 2012, Biochemistry.

[21]  W. Debinski,et al.  EphA2 as a Novel Molecular Marker and Target in Glioblastoma Multiforme , 2005, Molecular Cancer Research.

[22]  V. Preedy,et al.  European Organization for Research and Treatment of Cancer , 2010 .

[23]  J. Bolz,et al.  Ephrins regulate the formation of terminal axonal arbors during the development of thalamocortical projections. , 2002, Development.

[24]  I. Bayazitov,et al.  A perivascular niche for brain tumor stem cells. , 2007, Cancer cell.

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

[26]  Mark Bernstein,et al.  Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens. , 2009, Cell stem cell.

[27]  R. Henkelman,et al.  Identification of human brain tumour initiating cells , 2004, Nature.

[28]  J. García-Verdugo,et al.  Disruption of Eph/ephrin signaling affects migration and proliferation in the adult subventricular zone , 2000, Nature Neuroscience.

[29]  Martin J. van den Bent,et al.  Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. , 2005, The New England journal of medicine.

[30]  D. Douek,et al.  Isolation of viable antigen-specific CD8+ T cells based on membrane-bound tumor necrosis factor (TNF)-α expression. , 2011, Journal of immunological methods.

[31]  Ugo Orfanelli,et al.  Isolation and Characterization of Tumorigenic, Stem-like Neural Precursors from Human Glioblastoma , 2004, Cancer Research.

[32]  H. Xi,et al.  Aberrant expression of EphA3 in gastric carcinoma: correlation with tumor angiogenesis and survival , 2012, Journal of Gastroenterology.

[33]  Zang Ai-hua,et al.  Stem Cells,Cancer and Cancer Stem Cells , 2005 .

[34]  A. Boyd,et al.  Embryonic stem cells express multiple Eph-subfamily receptor tyrosine kinases. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[35]  I. Wicks,et al.  Molecular cloning of HEK, the gene encoding a receptor tyrosine kinase expressed by human lymphoid tumor cell lines. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[36]  M. Goulding,et al.  Expression of theTyro4/Mek4/Cek4Gene Specifically Marks a Subset of Embryonic Motor Neurons and Their Muscle Targets , 1996, Molecular and Cellular Neuroscience.

[37]  G. Lemke,et al.  A critical role for the EphA3 receptor tyrosine kinase in heart development. , 2007, Developmental biology.

[38]  J. Uhm,et al.  The transcriptional network for mesenchymal transformation of brain tumours , 2010 .

[39]  L. D. Ward,et al.  Isolation and characterization of a novel receptor-type protein tyrosine kinase (hek) from a human pre-B cell line. , 1992, The Journal of biological chemistry.

[40]  David T Scadden,et al.  Ephrin receptor, EphB4, regulates ES cell differentiation of primitive mammalian hemangioblasts, blood, cardiomyocytes, and blood vessels. , 2004, Blood.

[41]  Jane E. Visvader,et al.  Cells of origin in cancer , 2011, Nature.

[42]  Andrew D. Yates,et al.  Somatic mutations of the protein kinase gene family in human lung cancer. , 2005, Cancer research.

[43]  Yuan Qi,et al.  Integrated Genomic Analysis Identifies Clinically Relevant Subtypes of Glioblastoma Characterized by Abnormalities in PDGFRA , IDH 1 , EGFR , and NF 1 Citation Verhaak , 2010 .

[44]  J. Dick,et al.  Stem cell concepts renew cancer research. , 2008, Blood.

[45]  M. Berens,et al.  The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells. , 2004, Cancer research.

[46]  Thomas D. Wu,et al.  Molecular subclasses of high-grade glioma predict prognosis, delineate a pattern of disease progression, and resemble stages in neurogenesis. , 2006, Cancer cell.

[47]  D. Scadden,et al.  Ephrin receptor, EphB4, regulates ES cell differentiation of primitive mammalian hemangioblasts, blood, cardiomyocytes, and blood vessels. , 2004, Blood.

[48]  A. Boyd,et al.  Cloning and characterization of EphA3 (Hek) gene promoter: DNA methylation regulates expression in hematopoietic tumor cells. , 1999, Blood.

[49]  H. Fine,et al.  SSEA-1 is an enrichment marker for tumor-initiating cells in human glioblastoma. , 2009, Cell stem cell.