title: RAF265 inhibits the growth of advanced human melanoma tumors.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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 (BRAF(V600E/K)), whereas eight of 17 (47%) tumors were BRAF wild type (BRAF(WT)). 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 BRAF(WT), of which 1 carried c-KIT(L576P) and another N-RAS(Q61R) mutation, while only 2 (29%) of the responding tumors were BRAF(V600E/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.
authors.affiliation: Department of Veterans Affairs, Vanderbilt-Ingram Cancer Center and Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
authors.forename: Yingjun
authors.initials: Y
authors.lastname: Su
authors.forename: Anna E
authors.initials: AE
authors.lastname: Vilgelm
authors.forename: Mark C
authors.initials: MC
authors.lastname: Kelley
authors.forename: Oriana E
authors.initials: OE
authors.lastname: Hawkins
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Kelli L
authors.initials: KL
authors.lastname: Boyd
authors.forename: Sara
authors.initials: S
authors.lastname: Kantrow
authors.forename: Ryan C
authors.initials: RC
authors.lastname: Splittgerber
authors.forename: Sarah P
authors.initials: SP
authors.lastname: Short
authors.forename: Tammy
authors.initials: T
authors.lastname: Sobolik
authors.forename: Snjezana
authors.initials: S
authors.lastname: Zaja-Milatovic
authors.forename: Kimberly Brown
authors.initials: KB
authors.lastname: Dahlman
authors.forename: Katayoun I
authors.initials: KI
authors.lastname: Amiri
authors.forename: Aixiang
authors.initials: A
authors.lastname: Jiang
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Lu
authors.forename: Yu
authors.initials: Y
authors.lastname: Shyr
authors.forename: Darrin D
authors.initials: DD
authors.lastname: Stuart
authors.forename: Shawn
authors.initials: S
authors.lastname: Levy
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Sosman
authors.forename: Ann
authors.initials: A
authors.lastname: Richmond
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000096:biosynthesis; D049109:Cell Proliferation / Q000187:drug effects; D019938:Cyclin D1 / Q000096:biosynthesis; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology* / Q000627:therapeutic use*; D019394:Ki-67 Antigen / Q000096:biosynthesis; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008297:Male; D008545:Melanoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology / Q000556:secondary; D051379:Mice; D008819:Mice, Nude; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D011725:Pyridines / Q000494:pharmacology* / Q000627:therapeutic use*; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays; D051020:bcl-X Protein / Q000096:biosynthesis; D000097869:Polo-Like Kinase 1
chemical_list: D018797:Cell Cycle Proteins; D019938:Cyclin D1; D048049:Extracellular Signal-Regulated MAP Kinases; D007093:Imidazoles; D019394:Ki-67 Antigen; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D048493:Proto-Oncogene Proteins B-raf; D019009:Proto-Oncogene Proteins c-kit; D016283:Proto-Oncogene Proteins p21(ras); D011725:Pyridines; D051020:bcl-X Protein; D000097869:Polo-Like Kinase 1; C559019:RAF265; C482119:BRAF protein, human