title: MicroRNAs induced in melanoma treated with combination targeted therapy of Temsirolimus and Bevacizumab.
abstract: BACKGROUND
  Targeted therapies directed at commonly overexpressed pathways in melanoma have clinical activity in numerous trials. Little is known about how these therapies influence microRNA (miRNA) expression, particularly with combination regimens. Knowledge of miRNAs altered with treatment may contribute to understanding mechanisms of therapeutic effects, as well as mechanisms of tumor escape from therapy. We analyzed miRNA expression in metastatic melanoma tissue samples treated with a novel combination regimen of Temsirolimus and Bevacizumab. Given the preliminary clinical activity observed with this combination regimen, we hypothesized that we would see significant changes in miRNA expression with combination treatment.
  
  
  METHODS
  Using microarray analysis we analyzed miRNA expression levels in melanoma samples from a Cancer Therapy Evaluation Program-sponsored phase II trial of combination Temsirolimus and Bevacizumab in advanced melanoma, which elicited clinical benefit in a subset of patients. Pre-treatment and post-treatment miRNA levels were compared using paired t-tests between sample groups (patients), using a p-value < 0.01 for significance.
  
  
  RESULTS
  microRNA expression remained unchanged with Temsirolimus alone; however, expression of 15 microRNAs was significantly upregulated (1.4 to 2.5-fold) with combination treatment, compared to pre-treatment levels. Interestingly, twelve of these fifteen miRNAs possess tumor suppressor capabilities. We identified 15 putative oncogenes as potential targets of the 12 tumor suppressor miRNAs, based on published experimental evidence. For 15 of 25 miRNA-target mRNA pairings, changes in gene expression from pre-treatment to post-combination treatment samples were inversely correlated with changes in miRNA expression, supporting a functional effect of those miRNA changes. Clustering analyses based on selected miRNAs suggest preliminary signatures characteristic of clinical response to combination treatment and of tumor BRAF mutational status.
  
  
  CONCLUSIONS
  To our knowledge, this is the first study analyzing miRNA expression in pre-treatment and post-treatment human metastatic melanoma tissue samples. This preliminary investigation suggests miRNAs that may be involved in the mechanism of action of combination Temsirolimus and Bevacizumab in metastatic melanoma, possibly through inhibition of oncogenic pathways, and provides the preliminary basis for further functional studies of these miRNAs.
authors.affiliation: Department of Surgery, Division of Surgical Oncology, University of Virginia School of Medicine, Charlottesville, VA, USA. CLS8H@hscmail.mcc.virginia.edu.
authors.forename: Aubrey G
authors.initials: AG
authors.lastname: Wagenseller
authors.forename: Amber
authors.initials: A
authors.lastname: Shada
authors.forename: Kevin M
authors.initials: KM
authors.lastname: D'Auria
authors.forename: Cheryl
authors.initials: C
authors.lastname: Murphy
authors.forename: Dandan
authors.initials: D
authors.lastname: Sun
authors.forename: Kerrington R
authors.initials: KR
authors.lastname: Molhoek
authors.forename: Jason A
authors.initials: JA
authors.lastname: Papin
authors.forename: Anindya
authors.initials: A
authors.lastname: Dutta
authors.forename: Craig L
authors.initials: CL
authors.lastname: Slingluff
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology / Q000627:therapeutic use*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology / Q000627:therapeutic use; D000068258:Bevacizumab; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D058990:Molecular Targeted Therapy; D010865:Pilot Projects; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics; D015203:Reproducibility of Results; D020123:Sirolimus / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D035683:MicroRNAs; D000068258:Bevacizumab; C401859:temsirolimus; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D020123:Sirolimus
references.citation: Cancer Res. 2011 Aug 1;71(15):5144-53
references.pmid: 21670079
references.citation: Ann Surg Oncol. 2007 Aug;14(8):2367-76
references.pmid: 17534686
references.citation: Annu Rev Pathol. 2009;4:199-227
references.pmid: 18817506
references.citation: Nucleic Acids Res. 2005 Mar 01;33(4):1290-7
references.pmid: 15741182
references.citation: Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5116-21
references.pmid: 11309499
references.citation: Toxicol In Vitro. 2010 Apr;24(3):928-35
references.pmid: 19925859
references.citation: Melanoma Res. 2011 Jun;21(3):253-6
references.pmid: 21460750
references.citation: Clin Cancer Res. 2013 Jul 1;19(13):3611-20
references.pmid: 23620404
references.citation: Curr Opin Pharmacol. 2003 Aug;3(4):371-7
references.pmid: 12901945
references.citation: Mol Endocrinol. 2010 Feb;24(2):447-63
references.pmid: 20081105
references.citation: J Transl Med. 2005 Oct 28;3:39
references.pmid: 16255777
references.citation: Science. 2008 Apr 4;320(5872):97-100
references.pmid: 18292307
references.citation: Mol Cancer Res. 2010 Sep;8(9):1207-16
references.pmid: 20696752
references.citation: Nat Struct Mol Biol. 2009 Jan;16(1):23-9
references.pmid: 19079265
references.citation: Proc Natl Acad Sci U S A. 2011 Jan 11;108(2):522-7
references.pmid: 21187425
references.citation: Hepatology. 2010 Mar;51(3):836-45
references.pmid: 20041405
references.citation: J Immunol. 2005 Jun 1;174(11):6863-71
references.pmid: 15905528
references.citation: Cell Res. 2008 May;18(5):549-57
references.pmid: 18379589
references.citation: Cancer Immunol Immunother. 2000 Mar;48(12):661-72
references.pmid: 10752474
references.citation: Melanoma Res. 2011 Aug;21(4):274-84
references.pmid: 21654344
references.citation: Cell. 2005 Mar 11;120(5):635-47
references.pmid: 15766527
references.citation: Reprod Biol Endocrinol. 2012 Sep 06;10:72
references.pmid: 22950660
references.citation: Epigenetics. 2011 Mar;6(3):388-94
references.pmid: 21081840
references.citation: J Exp Clin Cancer Res. 2010 Nov 22;29:151
references.pmid: 21092188
references.citation: Mol Cancer. 2011 May 18;10:58
references.pmid: 21592394
references.citation: N Engl J Med. 2011 Jun 30;364(26):2507-16
references.pmid: 21639808
references.citation: Nat Med. 2002 Feb;8(2):128-35
references.pmid: 11821896
references.citation: Cancer Res. 2009 Aug 1;69(15):6275-81
references.pmid: 19584290
references.citation: Clin Exp Pharmacol Physiol. 2009 Sep;36(9):e32-9
references.pmid: 19473196
references.citation: J Virol. 2008 Sep;82(18):9065-74
references.pmid: 18596100
references.citation: J Control Release. 2011 Nov 7;155(3):427-34
references.pmid: 21723890
references.citation: Prostate. 2013 Jan;73(1):101-12
references.pmid: 22674191
references.citation: Cancer Res. 2004 Oct 1;64(19):7002-10
references.pmid: 15466193
references.citation: Trends Mol Med. 2010 Jun;16(6):257-67
references.pmid: 20493775
references.citation: Clin Exp Pharmacol Physiol. 2009 Feb;36(2):181-8
references.pmid: 18986336
references.citation: Cancer Res. 2011 Mar 1;71(5):1680-9
references.pmid: 21189327
references.citation: PLoS One. 2012;7(6):e38364
references.pmid: 22719881
references.citation: Cancer Res. 2010 Apr 15;70(8):3119-27
references.pmid: 20354188
references.citation: BMC Med Genomics. 2009 May 08;2:23
references.pmid: 19426511
references.citation: Gene. 2006 Dec 15;384:51-61
references.pmid: 16971064
references.citation: Int J Clin Exp Pathol. 2009 Mar 20;2(6):519-27
references.pmid: 19636399
references.citation: Cancer Res. 2011 Feb 15;71(4):1313-24
references.pmid: 21212412
references.citation: Cancer Res. 2010 Jun 15;70(12):5184-93
references.pmid: 20501828
references.citation: Cancer Res. 2007 Oct 15;67(20):9762-70
references.pmid: 17942906
references.citation: J Cutan Pathol. 2007 Aug;34(8):593-6
references.pmid: 17640227
references.citation: Genes Dev. 2007 May 1;21(9):1025-30
references.pmid: 17437991
references.citation: J Invest Dermatol. 2010 Aug;130(8):2062-70
references.pmid: 20357817
references.citation: J Cell Biol. 2010 Jun 28;189(7):1157-69
references.pmid: 20566686
references.citation: J Immunol. 1989 May 1;142(9):3329-35
references.pmid: 2785141
references.citation: Oncol Rep. 2011 Oct;26(4):1011-7
references.pmid: 21743970
references.citation: Cancer Res. 2008 Jun 1;68(11):4392-7
references.pmid: 18519701
references.citation: Cancer. 2005 Sep 1;104(5):1045-8
references.pmid: 16007689
references.citation: J Cell Physiol. 2009 Aug;220(2):485-91
references.pmid: 19391107
references.citation: J Invest Dermatol. 2009 May;129(5):1219-24
references.pmid: 19005486
references.citation: Carcinogenesis. 2012 Oct;33(10):1823-32
references.pmid: 22693259
references.citation: Nat Rev Cancer. 2006 Nov;6(11):857-66
references.pmid: 17060945
references.citation: J Pathol. 2011 Jan;223(2):241-50
references.pmid: 21125678
references.citation: Int J Cancer. 2011 Dec 15;129(12):2807-15
references.pmid: 21544800
references.citation: Melanoma Res. 2010 Dec;20(6):479-84
references.pmid: 20827223
references.citation: Hepatology. 2010 Nov;52(5):1731-40
references.pmid: 20827722