Combining molecular and immunohistochemical analyses of key drivers in primary melanomas: interplay between germline and somatic variations

Due to the high mutational somatic burden of Cutaneous Malignant Melanoma (CMM) a thorough profiling of the driver mutations and their interplay is necessary to explain the timing of tumorigenesis or for the identification of actionable genetic events. The aim of this study was to establish the mutation rate of some of the key drivers in melanoma tumorigenesis combining molecular analyses and/or immunohistochemistry in 93 primary CMMs from an Italian cohort also characterized for germline status, and to investigate an interplay between germline and somatic variants. BRAF mutations were present in 68% of cases, while CDKN2A germline mutations were found in 16 % and p16 loss in tissue was found in 63%. TERT promoter somatic mutations were detected in 38% of cases while the TERT –245T>C polymorphism was found in 51% of cases. NRAS mutations were found in 39% of BRAF negative or undetermined cases. NF1 was expressed in all cases analysed. MC1R variations were both considered as a dichotomous variable or scored. While a positive, although not significant association between CDKN2A germline mutations, but not MC1R variants, and BRAF somatic mutation was found, we did not observe other associations between germline and somatic events. A yet undescribed inverse correlation between TERT –245T>C polymorphism and the presence of BRAF mutation was found. It is possible to hypothesize that –245T>C polymorphism could be included in those genotypes which may influence the occurrence of BRAF mutations. Further studies are needed to investigate the role of –245T>C polymorphism as a germline predictor of BRAF somatic mutation status.

[1]  K. Hemminki,et al.  TERT promoter mutations in melanoma survival , 2019, Oncotarget.

[2]  R. Fiocca,et al.  Quality control in diagnostic immunohistochemistry: integrated on-slide positive controls , 2017, Histochemistry and Cell Biology.

[3]  K. Peris,et al.  Characterization of melanoma susceptibility genes in high-risk patients from Central Italy , 2017, Melanoma research.

[4]  H. Rizos,et al.  Mechanisms and strategies to overcome resistance to molecularly targeted therapy for melanoma , 2017, Cancer.

[5]  H. Tsao,et al.  Somatic driver mutations in melanoma , 2017, Cancer.

[6]  Catherine A. Shang,et al.  Whole-genome landscapes of major melanoma subtypes , 2017, Nature.

[7]  J. Kirkwood,et al.  Genetic and Epigenetic Alterations of TERT Are Associated with Inferior Outcome in Adolescent and Young Adult Patients with Melanoma , 2017, Scientific Reports.

[8]  Elizabeth Lazaridou,et al.  Epidemiological trends in skin cancer , 2017, Dermatology practical & conceptual.

[9]  Rajiv Kumar,et al.  TERT promoter mutations associate with MC1R variants in melanoma patients , 2017, Pigment cell & melanoma research.

[10]  N. Shen,et al.  Association between rs2853669 in TERT gene and the risk and prognosis of human cancer: a systematic review and meta-analysis. , 2017, Oncotarget.

[11]  Erik Larsson,et al.  Recurrent promoter mutations in melanoma are defined by an extended context-specific mutational signature , 2017, bioRxiv.

[12]  A. Ballestrero,et al.  Heterogeneity and frequency of BRAF mutations in primary melanoma: Comparison between molecular methods and immunohistochemistry , 2016, Oncotarget.

[13]  J. Becker,et al.  Distribution of TERT promoter mutations in primary and metastatic melanomas in Austrian patients , 2016, Journal of Cancer Research and Clinical Oncology.

[14]  Federica Grillo,et al.  A combination of immunohistochemistry and molecular approaches improves highly sensitive detection of BRAF mutations in papillary thyroid cancer , 2016, Endocrine.

[15]  M. Honavar,et al.  The prognostic impact of TERT promoter mutations in glioblastomas is modified by the rs2853669 single nucleotide polymorphism , 2016, International journal of cancer.

[16]  E. Martínez-Balibrea,et al.  Resistant mechanisms to BRAF inhibitors in melanoma. , 2016, Annals of translational medicine.

[17]  N. Hayward,et al.  Histologic and Phenotypic Factors and MC1R Status Associated with BRAF(V600E), BRAF(V600K), and NRAS Mutations in a Community-Based Sample of 414 Cutaneous Melanomas. , 2016, The Journal of investigative dermatology.

[18]  Rajiv Kumar,et al.  TERT promoter mutations associate with fast‐growing melanoma , 2016, Pigment cell & melanoma research.

[19]  P. Queirolo,et al.  The CDKN2A/p16INK4a 5′UTR sequence and translational regulation: impact of novel variants predisposing to melanoma , 2016, Pigment cell & melanoma research.

[20]  P. Ascierto,et al.  Multiple primary melanomas (MPMs) and criteria for genetic assessment: MultiMEL, a multicenter study of the Italian Melanoma Intergroup. , 2016, Journal of the American Academy of Dermatology.

[21]  G. Jung,et al.  The TERT promoter SNP rs2853669 decreases E2F1 transcription factor binding and increases mortality and recurrence risks in liver cancer , 2015, Oncotarget.

[22]  J. Wargo,et al.  Utility of BRAF V600E Immunohistochemistry Expression Pattern as a Surrogate of BRAF Mutation Status in 154 Patients with Advanced Melanoma. , 2015, Human pathology.

[23]  Steven J. M. Jones,et al.  Genomic Classification of Cutaneous Melanoma , 2015, Cell.

[24]  Chibo Hong,et al.  The transcription factor GABP selectively binds and activates the mutant TERT promoter in cancer , 2015, Science.

[25]  E. Nagore,et al.  BRAF, NRAS and MC1R status in a prospective series of primary cutaneous melanoma , 2015, The British journal of dermatology.

[26]  G. Fontanini,et al.  Immunohistochemistry is highly sensitive and specific for the detection of NRASQ61R mutation in melanoma , 2015, Modern Pathology.

[27]  R. Fiocca,et al.  Factors affecting immunoreactivity in long-term storage of formalin-fixed paraffin-embedded tissue sections , 2015, Histochemistry and Cell Biology.

[28]  M. Simoni,et al.  Telomerase in differentiated thyroid cancer: Promoter mutations, expression and localization , 2015, Molecular and Cellular Endocrinology.

[29]  M. Ringnér,et al.  Primary melanoma tumors from CDKN2A mutation carriers do not belong to a distinct molecular subclass. , 2014, The Journal of investigative dermatology.

[30]  E. Larsson,et al.  Systematic analysis of noncoding somatic mutations and gene expression alterations across 14 tumor types , 2014, Nature Genetics.

[31]  K. Hoang-Xuan,et al.  TERT promoter mutations in gliomas, genetic associations and clinico-pathological correlations , 2014, British Journal of Cancer.

[32]  D. Schadendorf,et al.  TERT promoter mutation status as an independent prognostic factor in cutaneous melanoma. , 2014, Journal of the National Cancer Institute.

[33]  Miguel Melo,et al.  Telomerase promoter mutations in cancer: an emerging molecular biomarker? , 2014, Virchows Archiv.

[34]  S. Hamilton-Dutoit,et al.  Immunohistochemical analysis of molecular drivers in melanoma identifies p16 as an independent prognostic biomarker , 2014, Journal of Clinical Pathology.

[35]  J. Becker,et al.  Telomerase reverse transcriptase promoter mutations in primary cutaneous melanoma. , 2014, Nature communications.

[36]  P. Queirolo,et al.  Prevalence of the E318K MITF germline mutation in Italian melanoma patients: associations with histological subtypes and family cancer history , 2013, Pigment cell & melanoma research.

[37]  D. Schadendorf,et al.  TERT Promoter Mutations in Familial and Sporadic Melanoma , 2013, Science.

[38]  P. Queirolo,et al.  MC1R variation and melanoma risk in relation to host/clinical and environmental factors in CDKN2A positive and negative melanoma patients , 2012, Experimental dermatology.

[39]  A. Sivachenko,et al.  A Landscape of Driver Mutations in Melanoma , 2012, Cell.

[40]  D. Schadendorf,et al.  Inherited variants in the MC1R gene and survival from cutaneous melanoma: a BioGenoMEL study , 2012, Pigment cell & melanoma research.

[41]  N. Hayward,et al.  Melanocortin 1 receptor and risk of cutaneous melanoma: A meta‐analysis and estimates of population burden , 2011, International journal of cancer.

[42]  R. Scolyer,et al.  BRAF mutations in cutaneous melanoma are independently associated with age, anatomic site of the primary tumor, and the degree of solar elastosis at the primary tumor site , 2011, Pigment cell & melanoma research.

[43]  J. Choi,et al.  Frequencies of BRAF and NRAS mutations are different in histological types and sites of origin of cutaneous melanoma: a meta‐analysis , 2011, The British journal of dermatology.

[44]  C. Begg,et al.  Relationship between germline MC1R variants and BRAF-mutant melanoma in a North Carolina population-based study. , 2010, The Journal of investigative dermatology.

[45]  P. Ascierto,et al.  Clinical genetic testing for familial melanoma in Italy: a cooperative study. , 2009, Journal of the American Academy of Dermatology.

[46]  A. Halpern,et al.  Selection criteria for genetic assessment of patients with familial melanoma. , 2009, Journal of the American Academy of Dermatology.

[47]  P. Queirolo,et al.  CDKN2A mutations and MC1R variants in Italian patients with single or multiple primary melanoma , 2008, Pigment cell & melanoma research.

[48]  D. Elder,et al.  MC1R variants increase risk of melanomas harboring BRAF mutations. , 2008, The Journal of investigative dermatology.

[49]  C. Pipper,et al.  [''R"--project for statistical computing]. , 2008, Ugeskrift for laeger.

[50]  A. Goldstein,et al.  Impact of E27X, a novel CDKN2A germ line mutation, on p16 and p14ARF expression in Italian melanoma families displaying pancreatic cancer and neuroblastoma. , 2006, Human molecular genetics.

[51]  D. Pinkel,et al.  MC1R germline variants confer risk for BRAF-mutant melanoma. , 2006, Science.

[52]  Stephen B Gruber,et al.  BRAF and NRAS mutations in melanoma and melanocytic nevi , 2006, Melanoma research.

[53]  A. Bosserhoff.  Novel biomarkers in malignant melanoma. , 2006, Clinica chimica acta; international journal of clinical chemistry.

[54]  M. Sormani,et al.  Early onset may predict G101W CDKN2A founder mutation carrier status in Ligurian melanoma patients , 2004, Melanoma research.

[55]  N. Hayward,et al.  Expression and localization of mutant p16 proteins in melanocytic lesions from familial melanoma patients. , 2004, Human pathology.

[56]  L. Akslen,et al.  Loss of nuclear p16 protein expression correlates with increased tumor cell proliferation (Ki-67) and poor prognosis in patients with vertical growth phase melanoma. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[57]  S. Puig,et al.  Somatic BRAF and NRAS mutations in familial melanomas with known germline CDKN2A status: a GenoMEL study. , 2014, The Journal of investigative dermatology.

[58]  D. Schadendorf,et al.  Highly Recurrent TERT Promoter Mutations in Human Melanoma , 2022 .