The tyrosine kinase inhibitor ZD6474 blocks proliferation of RET mutant medullary thyroid carcinoma cells.

Oncogenic conversion of the RET tyrosine kinase is a frequent feature of medullary thyroid carcinoma (MTC). ZD6474 (vandetanib) is an ATP-competitive inhibitor of RET, epidermal growth factor receptor (EGFR), and vascular endothelial growth factor receptors kinases. In this study, we have studied ZD6474 mechanism of action in TT and MZ-CRC-1 human MTC cell lines, carrying cysteine 634 to tryptophan (C634W) and methionine 918 to threonine (M918T) RET mutation respectively. ZD6474 blunted MTC cell proliferation and RET, Shc and p44/p42 mitogen-activated protein kinase (MAPK) phosphorylation. Single receptor knockdown by RNA interference showed that MTC cells depended on RET for proliferation. Adoptive expression of the ZD6474-resistant V804M RET mutant rescued proliferation of TT cells under ZD6474 treatment, showing that RET is a key ZD6474 target in these MTC cells. Upon RET inhibition, adoptive stimulation of EGFR partially rescued TT cell proliferation, MAPK signaling, and expression of cell-cycle-related genes. This suggests that simultaneous inhibition of RET and EGFR by ZD6474 may overcome the risk of MTC cells to escape from RET blockade through compensatory over-activation of EGFR.

[1]  G. Tortora,et al.  EGFR antagonists in cancer treatment. , 2008, The New England journal of medicine.

[2]  Keith L. Ligon,et al.  Coactivation of Receptor Tyrosine Kinases Affects the Response of Tumor Cells to Targeted Therapies , 2007, Science.

[3]  Y. Yarden.  The EGFR family and its ligands in human cancer. signalling mechanisms and therapeutic opportunities. , 2001, European journal of cancer.

[4]  Teresa Guida,et al.  The kinase inhibitor PP1 blocks tumorigenesis induced by RET oncogenes. , 2002, Cancer research.

[5]  A. Ryan,et al.  ZD6474 suppresses oncogenic RET isoforms in a Drosophila model for type 2 multiple endocrine neoplasia syndromes and papillary thyroid carcinoma. , 2005, Cancer research.

[6]  S. Asa,et al.  Expression of growth factors and growth factor receptors in normal and tumorous human thyroid tissues. , 1995, Thyroid : official journal of the American Thyroid Association.

[7]  Svend Kjær,et al.  Structure and Chemical Inhibition of the RET Tyrosine Kinase Domain* , 2006, Journal of Biological Chemistry.

[8]  K. Owzar,et al.  Prophylactic thyroidectomy in multiple endocrine neoplasia type 2A. , 2005, The New England journal of medicine.

[9]  O. Nilsson,et al.  Expression of transforming growth factor alpha and its receptor in human neuroendocrine tumours , 1995, International journal of cancer.

[10]  A. Knuth,et al.  Cytogenetic characterization of three human and three rat medullary thyroid carcinoma cell lines. , 1995, Cancer genetics and cytogenetics.

[11]  P. Michieli.  Hypoxia, angiogenesis and cancer therapy: To breathe or not to breathe? , 2009, Cell cycle.

[12]  M. Stridsberg,et al.  A transplantable human medullary thyroid carcinoma as a model for RET tyrosine kinase-driven tumorigenesis. , 2007, Endocrine-related cancer.

[13]  G. Fontanini,et al.  Antitumor Effects of ZD 6474 , a Small Molecule Vascular Endothelial Growth Factor Receptor Tyrosine Kinase Inhibitor , with Additional Activity against Epidermal Growth Factor Receptor Tyrosine Kinase 1 , 2003 .

[14]  S. Sherman.  Advances in chemotherapy of differentiated epithelial and medullary thyroid cancers. , 2009, The Journal of clinical endocrinology and metabolism.

[15]  Giampietro Gasparini,et al.  Combined targeting of EGFR-dependent and VEGF-dependent pathways: rationale, preclinical studies and clinical applications , 2008, Nature Clinical Practice Oncology.

[16]  J. Wass,et al.  Angiogenesis and lymphangiogenesis in thyroid proliferative lesions: relationship to type and tumour behaviour. , 2004, Endocrine-related cancer.

[17]  F. Raue,et al.  Prophylactic thyroidectomy in multiple endocrine neoplasia type 2 , 2010, Expert review of endocrinology & metabolism.

[18]  A. Vortmeyer,et al.  Amplification and overexpression of mutant RET in multiple endocrine neoplasia type 2-associated medullary thyroid carcinoma. , 2003, The Journal of clinical endocrinology and metabolism.

[19]  John M L Ebos,et al.  Accelerated metastasis after short-term treatment with a potent inhibitor of tumor angiogenesis. , 2009, Cancer cell.

[20]  G. Fontanini,et al.  Antitumor effects of ZD6474, a small molecule vascular endothelial growth factor receptor tyrosine kinase inhibitor, with additional activity against epidermal growth factor receptor tyrosine kinase. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[21]  A. Berns,et al.  Targeted anticancer therapies: mouse models help uncover the mechanisms of tumor escape. , 2008, Cancer cell.

[22]  D. Fabbro,et al.  RET/PTC-induced cell growth is mediated in part by epidermal growth factor receptor (EGFR) activation: evidence for molecular and functional interactions between RET and EGFR. , 2008, Cancer research.

[23]  E. Hayden.  Cutting off cancer's supply lines , 2009, Nature.

[24]  J. Cigudosa,et al.  Overexpression and activation of EGFR and VEGFR2 in medullary thyroid carcinomas is related to metastasis. , 2010, Endocrine-related cancer.

[25]  R. Herbst,et al.  Vandetanib (ZD6474): an orally available receptor tyrosine kinase inhibitor that selectively targets pathways critical for tumor growth and angiogenesis , 2007, Expert opinion on investigational drugs.

[26]  Y. Murakumo,et al.  RET receptor signaling: Dysfunction in thyroid cancer and Hirschsprung's disease , 2006, Pathology international.

[27]  E. Baudin,et al.  Vandetanib (VAN) in locally advanced or metastatic medullary thyroid cancer (MTC): A randomized, double-blind phase III trial (ZETA). , 2010 .

[28]  G. Fontanini,et al.  ZD6474, an orally available inhibitor of KDR tyrosine kinase activity, efficiently blocks oncogenic RET kinases. , 2002, Cancer research.

[29]  E. Baudin,et al.  New therapeutic approaches to treat medullary thyroid carcinoma , 2008, Nature Clinical Practice Endocrinology &Metabolism.

[30]  M. Santoro,et al.  Targeting the RET Pathway in Thyroid Cancer , 2009, Clinical Cancer Research.

[31]  L. Paz-Ares,et al.  Vandetanib (100 mg) in patients with locally advanced or metastatic hereditary medullary thyroid cancer. , 2010, The Journal of clinical endocrinology and metabolism.

[32]  A. Ryan,et al.  Vascular Endothelial Growth Factor Receptor-1 Contributes to Resistance to Anti–Epidermal Growth Factor Receptor Drugs in Human Cancer Cells , 2008, Clinical Cancer Research.

[33]  A. Ryan,et al.  Disease associated mutations at valine 804 in the RET receptor tyrosine kinase confer resistance to selective kinase inhibitors , 2004, Oncogene.

[34]  D. Pfister,et al.  Harvesting the low-hanging fruit: kinase inhibitors for therapy of advanced medullary and nonmedullary thyroid cancer. , 2010, The Journal of clinical endocrinology and metabolism.

[35]  M. Santoro,et al.  Drug Insight: small-molecule inhibitors of protein kinases in the treatment of thyroid cancer , 2006, Nature Clinical Practice Endocrinology &Metabolism.

[36]  M. Santoro,et al.  Point mutation of the RET proto-oncogene in the TT human medullary thyroid carcinoma cell line. , 1995, Biochemical and biophysical research communications.

[37]  A. Pinchera,et al.  Prognostic significance of somatic RET oncogene mutations in sporadic medullary thyroid cancer: a 10-year follow-up study. , 2008, The Journal of clinical endocrinology and metabolism.

[38]  J. Baselga,et al.  Targeting Tyrosine Kinases in Cancer: The Second Wave , 2006, Science.

[39]  Sonja Loges,et al.  Silencing or fueling metastasis with VEGF inhibitors: antiangiogenesis revisited. , 2009, Cancer cell.

[40]  Masahiro Inoue,et al.  Antiangiogenic therapy elicits malignant progression of tumors to increased local invasion and distant metastasis. , 2009, Cancer cell.

[41]  B. Curry,et al.  ZD6474 inhibits vascular endothelial growth factor signaling, angiogenesis, and tumor growth following oral administration. , 2002, Cancer research.

[42]  K. Shokat,et al.  Targeting the cancer kinome through polypharmacology , 2010, Nature Reviews Cancer.