RAF265 Inhibits the Growth of Advanced Human Melanoma Tumors

Purpose: The purpose of this preclinical study was to determine the effectiveness of RAF265, a multikinase inhibitor, for treatment of human metastatic melanoma and to characterize traits associated with drug response. Experimental Design: Advanced metastatic melanoma tumors from 34 patients were orthotopically implanted to nude mice. Tumors that grew in mice (17 of 34) were evaluated for response to RAF265 (40 mg/kg, every day) over 30 days. The relation between patient characteristics, gene mutation profile, global gene expression profile, and RAF265 effects on tumor growth, mitogen-activated protein/extracellular signal-regulated kinase (MEK)/extracellular signal-regulated kinase (ERK) phosphorylation, proliferation, and apoptosis markers was evaluated. Results: Nine of the 17 tumors that successfully implanted (53%) were mutant BRAF (BRAFV600E/K), whereas eight of 17 (47%) tumors were BRAF wild type (BRAFWT). Tumor implants from 7 of 17 patients (41%) responded to RAF265 treatment with more than 50% reduction in tumor growth. Five of the 7 (71%) responders were BRAFWT, of which 1 carried c-KITL576P and another N-RASQ61R mutation, while only 2 (29%) of the responding tumors were BRAFV600E/K. Gene expression microarray data from nonimplanted tumors revealed that responders exhibited enriched expression of genes involved in cell growth, proliferation, development, cell signaling, gene expression, and cancer pathways. Although response to RAF265 did not correlate with pERK1/2 reduction, RAF265 responders did exhibit reduced pMEK1, reduced proliferation based upon reduced Ki-67, cyclin D1 and polo-like kinase1 levels, and induction of the apoptosis mediator BCL2-like 11. Conclusions: Orthotopic implants of patient tumors in mice may predict prognosis and treatment response for melanoma patients. A subpopulation of human melanoma tumors responds to RAF265 and can be characterized by gene mutation and gene expression profiles. Clin Cancer Res; 18(8); 2184–98. ©2012 AACR.

[1]  Correction: RAF265 Inhibits the Growth of Advanced Human Melanoma Tumors , 2012, Clinical Cancer Research.

[2]  O. Feron,et al.  Lactate shuttles at a glance: from physiological paradigms to anti-cancer treatments , 2011, Disease Models & Mechanisms.

[3]  J. Martens,et al.  Mesenchymal stem cells and carcinoma-associated fibroblasts sensitize breast cancer cells in 3D cultures to kinase inhibitors. , 2011, International journal of oncology.

[4]  Donald M Miller,et al.  Novel Immunotherapeutic Agents and Small Molecule Antagonists of Signalling Kinases for the Treatment of Metastatic Melanoma , 2011, Drugs.

[5]  A. Hauschild,et al.  Improved survival with vemurafenib in melanoma with BRAF V600E mutation. , 2011, The New England journal of medicine.

[6]  M. Labow,et al.  Protein kinase D3 sensitizes RAF inhibitor RAF265 in melanoma cells by preventing reactivation of MAPK signaling. , 2011, Cancer research.

[7]  David E Fisher,et al.  New Strategies in Metastatic Melanoma: Oncogene-Defined Taxonomy Leads to Therapeutic Advances , 2011, Clinical Cancer Research.

[8]  J. Iovanna,et al.  Nupr1: The Swiss‐knife of cancer , 2011, Journal of cellular physiology.

[9]  K. Flaherty,et al.  Results from the first-in-human (FIH) phase I study of the oral RAF inhibitor RAF265 administered daily to patients with advanced cutaneous melanoma. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[10]  S. Gambhir,et al.  Use of DNA microarray and small animal positron emission tomography in preclinical drug evaluation of RAF265, a novel B-Raf/VEGFR-2 inhibitor. , 2011, Neoplasia.

[11]  C. Der.  Faculty Opinions recommendation of COT drives resistance to RAF inhibition through MAP kinase pathway reactivation. , 2010 .

[12]  A. Aplin,et al.  The wrath of RAFs: rogue behavior of B-RAF kinase inhibitors. , 2010, The Journal of investigative dermatology.

[13]  Kam Y. J. Zhang,et al.  Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF-mutant melanoma , 2010, Nature.

[14]  P. Hersey,et al.  Apoptosis of human melanoma cells induced by inhibition of B-RAFV600E involves preferential splicing of bimS , 2010, Cell Death and Disease.

[15]  K. Flaherty,et al.  Inhibition of mutated, activated BRAF in metastatic melanoma. , 2010, The New England journal of medicine.

[16]  D. Schadendorf,et al.  Improved survival with ipilimumab in patients with metastatic melanoma. , 2010, The New England journal of medicine.

[17]  A. Borczuk,et al.  Dual specificity phosphatase 6 (DUSP6) is an ETS-regulated negative feedback mediator of oncogenic ERK signaling in lung cancer cells. , 2010, Carcinogenesis.

[18]  M. Belvin,et al.  RAF inhibitors prime wild-type RAF to activate the MAPK pathway and enhance growth , 2010, Nature.

[19]  Chao Zhang,et al.  RAF inhibitors transactivate RAF dimers and ERK signaling in cells with wild-type BRAF , 2010, Nature.

[20]  B. Taylor,et al.  Transcriptional pathway signatures predict MEK addiction and response to selumetinib (AZD6244). , 2010, Cancer research.

[21]  J. Reis-Filho,et al.  Kinase-Dead BRAF and Oncogenic RAS Cooperate to Drive Tumor Progression through CRAF , 2010, Cell.

[22]  Kou-Juey Wu,et al.  Regulation of membrane-type 4 matrix metalloproteinase by SLUG contributes to hypoxia-mediated metastasis. , 2009, Neoplasia.

[23]  Nathanael Gray,et al.  Factors underlying sensitivity of cancers to small-molecule kinase inhibitors , 2009, Nature Reviews Drug Discovery.

[24]  S. Kondo,et al.  Phosphorylated ezrin is associated with EBV latent membrane protein 1 in nasopharyngeal carcinoma and induces cell migration , 2009, Oncogene.

[25]  A. Strasser,et al.  Treatment of B-RAF mutant human tumor cells with a MEK inhibitor requires Bim and is enhanced by a BH3 mimetic. , 2008, The Journal of clinical investigation.

[26]  Abran Q. Costales,et al.  Design and synthesis of orally bioavailable benzimidazoles as Raf kinase inhibitors. , 2008, Journal of medicinal chemistry.

[27]  Wei Liu,et al.  The melanoma-associated antigen A3 mediates fibronectin-controlled cancer progression and metastasis. , 2008, Cancer research.

[28]  G. Sommer,et al.  Reference , 2008 .

[29]  Robert C. Burghardt,et al.  Loss of Singleminded-2s in the Mouse Mammary Gland Induces an Epithelial-Mesenchymal Transition Associated with Up-Regulation of Slug and Matrix Metalloprotease 2 , 2007, Molecular and Cellular Biology.

[30]  Zhiwei Wang,et al.  Down-regulation of platelet-derived growth factor-D inhibits cell growth and angiogenesis through inactivation of notch-1 and nuclear factor-κB signaling , 2007 .

[31]  A. Simpson,et al.  MAGE-A, mMage-b, and MAGE-C proteins form complexes with KAP1 and suppress p53-dependent apoptosis in MAGE-positive cell lines. , 2007, Cancer research.

[32]  Suzanne Cory,et al.  Bcl-2-regulated apoptosis: mechanism and therapeutic potential. , 2007, Current opinion in immunology.

[33]  D. Stupack,et al.  The CD44 standard/ezrin complex regulates Fas-mediated apoptosis in Jurkat cells , 2007, Apoptosis.

[34]  D. Steinbach,et al.  PRAME expression is not associated with down-regulation of retinoic acid signaling in primary acute myeloid leukemia. , 2007, Cancer genetics and cytogenetics.

[35]  C. Nerlov,et al.  Cdk6 blocks myeloid differentiation by interfering with Runx1 DNA binding and Runx1‐C/EBPα interaction , 2007, The EMBO journal.

[36]  N. Hayward,et al.  Confirmation of a BRAF mutation-associated gene expression signature in melanoma. , 2007, Pigment cell research.

[37]  H. Ditzel,et al.  MAGE-A1, GAGE and NY-ESO-1 cancer/testis antigen expression during human gonadal development. , 2007, Human reproduction.

[38]  M. Mason,et al.  Does the PGC-1/PPARgamma pathway play a role in Com-1/p8 mediated cell growth inhibition in prostate cancer? , 2006, International journal of molecular medicine.

[39]  W. Jiang,et al.  Com-1/p8 acts as a putative tumour suppressor in prostate cancer. , 2006, International journal of molecular medicine.

[40]  M. Ratain,et al.  Sorafenib in advanced melanoma: a Phase II randomised discontinuation trial analysis , 2006, British Journal of Cancer.

[41]  M. Pierotti,et al.  MAGE-A tumor antigens target p53 transactivation function through histone deacetylase recruitment and confer resistance to chemotherapeutic agents. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[42]  Wei Zhang,et al.  Characterization of a novel Raf kinase inhibitor that causes target dependent tumor regression in human melanoma xenografts expressing mutant B-Raf , 2006 .

[43]  D. Barford,et al.  Wild-type and mutant B-RAF activate C-RAF through distinct mechanisms involving heterodimerization. , 2005, Molecular cell.

[44]  J. Fridlyand,et al.  Distinct sets of genetic alterations in melanoma. , 2005, The New England journal of medicine.

[45]  M. Katoh,et al.  Comparative genomics on SNAI1, SNAI2, and SNAI3 orthologs. , 2005, Oncology reports.

[46]  R. Taichman,et al.  Clusterin inhibits apoptosis by interacting with activated Bax , 2005, Nature Cell Biology.

[47]  C. Marshall,et al.  B-RAF is a therapeutic target in melanoma , 2004, Oncogene.

[48]  John N Weinstein,et al.  Predicting drug sensitivity and resistance: profiling ABC transporter genes in cancer cells. , 2004, Cancer cell.

[49]  K. Kinzler,et al.  Cancer genes and the pathways they control , 2004, Nature Medicine.

[50]  Markus Ringnér,et al.  Microarray expression profiling in melanoma reveals a BRAF mutation signature , 2004, Oncogene.

[51]  E. Fearon,et al.  The SLUG zinc-finger protein represses E-cadherin in breast cancer. , 2002, Cancer research.

[52]  J. Dagorn,et al.  p8 is critical for tumour development induced by rasV12 mutated protein and E1A oncogene , 2002, EMBO reports.

[53]  B. Edgar,et al.  Drosophila Cdk4 is required for normal growth and is dispensable for cell cycle progression , 2000, The EMBO journal.

[54]  W. Jiang,et al.  Ezrin regulates cell-cell and cell-matrix adhesion, a possible role with E-cadherin/beta-catenin. , 1999, Journal of cell science.

[55]  A. Iafrate,et al.  A platform for rapid detection of multiple oncogenic mutations with relevance to targeted therapy in non-small-cell lung cancer. , 2011, The Journal of molecular diagnostics : JMD.

[56]  春木 茂男.  Frequent silencing of protocadherin 17, a candidate tumour suppressor for esophageal squamous cell carcinoma , 2010 .