Human tumor genomics and zebrafish modeling identify SPRED1 loss as a driver of mucosal melanoma
Hunting and fishing for cancer genes
Mucosal melanoma is a rare, but deadly, form of melanoma that occurs in sun-protected tissues. Little is known about the genetic alterations that drive the growth of these tumors. Ablain et al. sequenced mucosal melanomas from 43 patients and found that a substantial fraction showed inactivation or loss of SPRED1, a gene that encodes a negative regulator of RAS–MAPK (mitogen-activated protein kinase) signaling. Using a platform called MAZERATI (Modeling Approach in Zebrafish for Rapid Tumor Initiation), they discovered that SPRED1 loss may help explain the poor response of melanoma patients to drugs that inhibit the KIT tyrosine kinase. The results suggest that a combination of KIT inhibitors and drugs that inhibit MAPK signaling may be more effective.
Science, this issue p. 1055
Abstract
Melanomas originating from mucosal surfaces have low mutation burden, genomic instability, and poor prognosis. To identify potential driver genes, we sequenced hundreds of cancer-related genes in 43 human mucosal melanomas, cataloging point mutations, amplifications, and deletions. The SPRED1 gene, which encodes a negative regulator of mitogen-activated protein kinase (MAPK) signaling, was inactivated in 37% of the tumors. Four distinct genotypes were associated with SPRED1 loss. Using a rapid, tissue-specific CRISPR technique to model these genotypes in zebrafish, we found that SPRED1 functions as a tumor suppressor, particularly in the context of KIT mutations. SPRED1 knockdown caused MAPK activation, increased cell proliferation, and conferred resistance to drugs inhibiting KIT tyrosine kinase activity. These findings provide a rationale for MAPK inhibition in SPRED1-deficient melanomas and introduce a zebrafish modeling approach that can be used more generally to dissect genetic interactions in cancer.
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Supplementary Material
Summary
Materials and Methods
Figs. S1 to S12
Tables S1 to S4
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References and Notes
1
F. G. Zak, W. Lawson, The presence of melanocytes in the nasal cavity. Ann. Otol. Rhinol. Laryngol. 83, 515–519 (1974).
2
O. J. Clemmensen, C. Fenger, Melanocytes in the anal canal epithelium. Histopathology 18, 237–241 (1991).
3
A. W. Barrett, A. M. H. Raja, The immunohistochemical identification of human oral mucosal melanocytes. Arch. Oral Biol. 42, 77–81 (1997).
4
S. J. Furney, S. Turajlic, G. Stamp, M. Nohadani, A. Carlisle, J. M. Thomas, A. Hayes, D. Strauss, M. Gore, J. van den Oord, J. Larkin, R. Marais, Genome sequencing of mucosal melanomas reveals that they are driven by distinct mechanisms from cutaneous melanoma. J. Pathol. 230, 261–269 (2013).
5
M. Krauthammer, Y. Kong, B. H. Ha, P. Evans, A. Bacchiocchi, J. P. McCusker, E. Cheng, M. J. Davis, G. Goh, M. Choi, S. Ariyan, D. Narayan, K. Dutton-Regester, A. Capatana, E. C. Holman, M. Bosenberg, M. Sznol, H. M. Kluger, D. E. Brash, D. F. Stern, M. A. Materin, R. S. Lo, S. Mane, S. Ma, K. K. Kidd, N. K. Hayward, R. P. Lifton, J. Schlessinger, T. J. Boggon, R. Halaban, Exome sequencing identifies recurrent somatic RAC1 mutations in melanoma. Nat. Genet. 44, 1006–1014 (2012).
6
J. A. Curtin, J. Fridlyand, T. Kageshita, H. N. Patel, K. J. Busam, H. Kutzner, K.-H. Cho, S. Aiba, E.-B. Bröcker, P. E. LeBoit, D. Pinkel, B. C. Bastian, Distinct sets of genetic alterations in melanoma. N. Engl. J. Med. 353, 2135–2147 (2005).
7
J. A. Curtin, K. Busam, D. Pinkel, B. C. Bastian, Somatic activation of KIT in distinct subtypes of melanoma. J. Clin. Oncol. 24, 4340–4346 (2006).
8
K. Byrd-Miles, E. L. Toombs, G. L. Peck, Skin cancer in individuals of African, Asian, Latin-American, and American-Indian descent: Differences in incidence, clinical presentation, and survival compared to Caucasians. J. Drugs Dermatol. 6, 10–16 (2007).
9
Z. Chi, S. Li, X. Sheng, L. Si, C. Cui, M. Han, J. Guo, Clinical presentation, histology, and prognoses of malignant melanoma in ethnic Chinese: A study of 522 consecutive cases. BMC Cancer 11, 85 (2011).
10
L. Altieri, M. K. Wong, D. H. Peng, M. Cockburn, Mucosal melanomas in the racially diverse population of California. J. Am. Acad. Dermatol. 76, 250–257 (2017).
11
D. D. Kirchoff, G. B. Deutsch, L. J. Foshag, J. H. Lee, M. S. Sim, M. B. Faries, Evolving therapeutic strategies in mucosal melanoma have not improved survival over five decades. Am. Surg. 82, 1–5 (2016).
12
E. Hodis, I. R. Watson, G. V. Kryukov, S. T. Arold, M. Imielinski, J.-P. Theurillat, E. Nickerson, D. Auclair, L. Li, C. Place, D. Dicara, A. H. Ramos, M. S. Lawrence, K. Cibulskis, A. Sivachenko, D. Voet, G. Saksena, N. Stransky, R. C. Onofrio, W. Winckler, K. Ardlie, N. Wagle, J. Wargo, K. Chong, D. L. Morton, K. Stemke-Hale, G. Chen, M. Noble, M. Meyerson, J. E. Ladbury, M. A. Davies, J. E. Gershenwald, S. N. Wagner, D. S. B. Hoon, D. Schadendorf, E. S. Lander, S. B. Gabriel, G. Getz, L. A. Garraway, L. Chin, A landscape of driver mutations in melanoma. Cell 150, 251–263 (2012).
13
J. Guo, L. Si, Y. Kong, K. T. Flaherty, X. Xu, Y. Zhu, C. L. Corless, L. Li, H. Li, X. Sheng, C. Cui, Z. Chi, S. Li, M. Han, L. Mao, X. Lin, N. Du, X. Zhang, J. Li, B. Wang, S. Qin, Phase II, open-label, single-arm trial of imatinib mesylate in patients with metastatic melanoma harboring c-Kit mutation or amplification. J. Clin. Oncol. 29, 2904–2909 (2011).
14
R. D. Carvajal, C. R. Antonescu, J. D. Wolchok, P. B. Chapman, R. A. Roman, J. Teitcher, K. S. Panageas, K. J. Busam, B. Chmielowski, J. Lutzky, A. C. Pavlick, A. Fusco, L. Cane, N. Takebe, S. Vemula, N. Bouvier, B. C. Bastian, G. K. Schwartz, KIT as a therapeutic target in metastatic melanoma. JAMA 305, 2327–2334 (2011).
15
F. S. Hodi, C. L. Corless, A. Giobbie-Hurder, J. A. Fletcher, M. Zhu, A. Marino-Enriquez, P. Friedlander, R. Gonzalez, J. S. Weber, T. F. Gajewski, S. J. O’Day, K. B. Kim, D. Lawrence, K. T. Flaherty, J. J. Luke, F. A. Collichio, M. S. Ernstoff, M. C. Heinrich, C. Beadling, K. A. Zukotynski, J. T. Yap, A. D. Van den Abbeele, G. D. Demetri, D. E. Fisher, Imatinib for melanomas harboring mutationally activated or amplified KIT arising on mucosal, acral, and chronically sun-damaged skin. J. Clin. Oncol. 31, 3182–3190 (2013).
16
Materials and methods are available as supplementary materials.
17
Cancer Genome Atlas Network, Genomic classification of cutaneous melanoma. Cell 161, 1681–1696 (2015).
18
P. T. C. Wan, M. J. Garnett, S. M. Roe, S. Lee, D. Niculescu-Duvaz, V. M. Good, C. M. Jones, C. J. Marshall, C. J. Springer, D. Barford, R. Marais; Cancer Genome Project, Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF. Cell 116, 855–867 (2004).
19
K. B. Dahlman, J. Xia, K. Hutchinson, C. Ng, D. Hucks, P. Jia, M. Atefi, Z. Su, S. Branch, P. L. Lyle, D. J. Hicks, V. Bozon, J. A. Glaspy, N. Rosen, D. B. Solit, J. L. Netterville, C. L. Vnencak-Jones, J. A. Sosman, A. Ribas, Z. Zhao, W. Pao, BRAFL597 mutations in melanoma are associated with sensitivity to MEK inhibitors. Cancer Discov. 2, 791–797 (2012).
20
H. Davies, G. R. Bignell, C. Cox, P. Stephens, S. Edkins, S. Clegg, J. Teague, H. Woffendin, M. J. Garnett, W. Bottomley, N. Davis, E. Dicks, R. Ewing, Y. Floyd, K. Gray, S. Hall, R. Hawes, J. Hughes, V. Kosmidou, A. Menzies, C. Mould, A. Parker, C. Stevens, S. Watt, S. Hooper, R. Wilson, H. Jayatilake, B. A. Gusterson, C. Cooper, J. Shipley, D. Hargrave, K. Pritchard-Jones, N. Maitland, G. Chenevix-Trench, G. J. Riggins, D. D. Bigner, G. Palmieri, A. Cossu, A. Flanagan, A. Nicholson, J. W. C. Ho, S. Y. Leung, S. T. Yuen, B. L. Weber, H. F. Seigler, T. L. Darrow, H. Paterson, R. Marais, C. J. Marshall, R. Wooster, M. R. Stratton, P. A. Futreal, Mutations of the BRAF gene in human cancer. Nature 417, 949–954 (2002).
21
T. Wakioka, A. Sasaki, R. Kato, T. Shouda, A. Matsumoto, K. Miyoshi, M. Tsuneoka, S. Komiya, R. Baron, A. Yoshimura, Spred is a Sprouty-related suppressor of Ras signalling. Nature 412, 647–651 (2001).
22
A. Nonami, R. Kato, K. Taniguchi, D. Yoshiga, T. Taketomi, S. Fukuyama, M. Harada, A. Sasaki, A. Yoshimura, Spred-1 negatively regulates interleukin-3-mediated ERK/mitogen-activated protein (MAP) kinase activation in hematopoietic cells. J. Biol. Chem. 279, 52543–52551 (2004).
23
I. B. Stowe, E. L. Mercado, T. R. Stowe, E. L. Bell, J. A. Oses-Prieto, H. Hernández, A. L. Burlingame, F. McCormick, A shared molecular mechanism underlies the human rasopathies Legius syndrome and neurofibromatosis-1. Genes Dev. 26, 1421–1426 (2012).
24
T. Dunzendorfer-Matt, E. L. Mercado, K. Maly, F. McCormick, K. Scheffzek, The neurofibromin recruitment factor Spred1 binds to the GAP related domain without affecting Ras inactivation. Proc. Natl. Acad. Sci. U.S.A. 113, 7497–7502 (2016).
25
Y. Hirata, H. Brems, M. Suzuki, M. Kanamori, M. Okada, R. Morita, I. Llano-Rivas, T. Ose, L. Messiaen, E. Legius, A. Yoshimura, Interaction between a domain of the negative regulator of the Ras-ERK pathway, SPRED1 protein, and the GTPase-activating protein-related domain of neurofibromin is implicated in Legius syndrome and neurofibromatosis type 1. J. Biol. Chem. 291, 3124–3134 (2016).
26
C. J. Ceol, Y. Houvras, J. Jane-Valbuena, S. Bilodeau, D. A. Orlando, V. Battisti, L. Fritsch, W. M. Lin, T. J. Hollmann, F. Ferré, C. Bourque, C. J. Burke, L. Turner, A. Uong, L. A. Johnson, R. Beroukhim, C. H. Mermel, M. Loda, S. Ait-Si-Ali, L. A. Garraway, R. A. Young, L. I. Zon, The histone methyltransferase SETDB1 is recurrently amplified in melanoma and accelerates its onset. Nature 471, 513–517 (2011).
27
J. Ablain, E. M. Durand, S. Yang, Y. Zhou, L. I. Zon, A CRISPR/Cas9 vector system for tissue-specific gene disruption in zebrafish. Dev. Cell 32, 756–764 (2015).
28
R. M. White, A. Sessa, C. Burke, T. Bowman, J. LeBlanc, C. Ceol, C. Bourque, M. Dovey, W. Goessling, C. E. Burns, L. I. Zon, Transparent adult zebrafish as a tool for in vivo transplantation analysis. Cell Stem Cell 2, 183–189 (2008).
29
F. W. Huang, E. Hodis, M. J. Xu, G. V. Kryukov, L. Chin, L. A. Garraway, Highly recurrent TERT promoter mutations in human melanoma. Science 339, 957–959 (2013).
30
I. Yeh, M. K. Tee, T. Botton, A. H. Shain, A. J. Sparatta, A. Gagnon, S. S. Vemula, M. C. Garrido, K. Nakamaru, T. Isoyama, T. H. McCalmont, P. E. LeBoit, B. C. Bastian, NTRK3 kinase fusions in Spitz tumours. J. Pathol. 240, 282–290 (2016).
31
L. Oesper, G. Satas, B. J. Raphael, Quantifying tumor heterogeneity in whole-genome and whole-exome sequencing data. Bioinformatics 30, 3532–3540 (2014).
32
K. Cibulskis, M. S. Lawrence, S. L. Carter, A. Sivachenko, D. Jaffe, C. Sougnez, S. Gabriel, M. Meyerson, E. S. Lander, G. Getz, Sensitive detection of somatic point mutations in impure and heterogeneous cancer samples. Nat. Biotechnol. 31, 213–219 (2013).
33
E. Talevich, A. H. Shain, T. Botton, B. C. Bastian, CNVkit: Genome-wide copy number detection and visualization from targeted DNA sequencing. PLOS Comput. Biol. 12, e1004873 (2016).
34
E. Garrison, G. Marth, Haplotype-based variant detection from short-read sequencing. arXiv:1207.3907 [q-bio.GN] (17 July 2012).
35
NHLBI GO Exome Sequencing Project (ESP), Exome Variant Server (Seattle); http://evs.gs.washington.edu/EVS/.
36
M. Lek, K. J. Karczewski, E. V. Minikel, K. E. Samocha, E. Banks, T. Fennell, A. H. O’Donnell-Luria, J. S. Ware, A. J. Hill, B. B. Cummings, T. Tukiainen, D. P. Birnbaum, J. A. Kosmicki, L. E. Duncan, K. Estrada, F. Zhao, J. Zou, E. Pierce-Hoffman, J. Berghout, D. N. Cooper, N. Deflaux, M. DePristo, R. Do, J. Flannick, M. Fromer, L. Gauthier, J. Goldstein, N. Gupta, D. Howrigan, A. Kiezun, M. I. Kurki, A. L. Moonshine, P. Natarajan, L. Orozco, G. M. Peloso, R. Poplin, M. A. Rivas, V. Ruano-Rubio, S. A. Rose, D. M. Ruderfer, K. Shakir, P. D. Stenson, C. Stevens, B. P. Thomas, G. Tiao, M. T. Tusie-Luna, B. Weisburd, H.-H. Won, D. Yu, D. M. Altshuler, D. Ardissino, M. Boehnke, J. Danesh, S. Donnelly, R. Elosua, J. C. Florez, S. B. Gabriel, G. Getz, S. J. Glatt, C. M. Hultman, S. Kathiresan, M. Laakso, S. McCarroll, M. I. McCarthy, D. McGovern, R. McPherson, B. M. Neale, A. Palotie, S. M. Purcell, D. Saleheen, J. M. Scharf, P. Sklar, P. F. Sullivan, J. Tuomilehto, M. T. Tsuang, H. C. Watkins, J. G. Wilson, M. J. Daly, D. G. MacArthur; Exome Aggregation Consortium, Analysis of protein-coding genetic variation in 60,706 humans. Nature 536, 285–291 (2016).
37
G. R. Abecasis, D. Altshuler, A. Auton, L. D. Brooks, R. M. Durbin, R. A. Gibbs, M. E. Hurles, G. A. McVean; 1000 Genomes Project Consortium, A map of human genome variation from population-scale sequencing. Nature 467, 1061–1073 (2010).
38
C. H. Mermel, S. E. Schumacher, B. Hill, M. L. Meyerson, R. Beroukhim, G. Getz, GISTIC2.0 facilitates sensitive and confident localization of the targets of focal somatic copy-number alteration in human cancers. Genome Biol. 12, R41 (2011).
39
J. L. Hartley, G. F. Temple, M. A. Brasch, DNA cloning using in vitro site-specific recombination. Genome Res. 10, 1788–1795 (2000).
40
O. Shalem, N. E. Sanjana, E. Hartenian, X. Shi, D. A. Scott, T. Mikkelson, D. Heckl, B. L. Ebert, D. E. Root, J. G. Doench, F. Zhang, Genome-scale CRISPR-Cas9 knockout screening in human cells. Science 343, 84–87 (2014).
41
K. Kawakami, H. Takeda, N. Kawakami, M. Kobayashi, N. Matsuda, M. Mishina, A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish. Dev. Cell 7, 133–144 (2004).
42
J. L. Maldonado, J. Fridlyand, H. Patel, A. N. Jain, K. Busam, T. Kageshita, T. Ono, D. G. Albertson, D. Pinkel, B. C. Bastian, Determinants of BRAF mutations in primary melanomas. J. Natl. Cancer Inst. 95, 1878–1890 (2003).
43
B. M. Helmke, J. Mollenhauer, C. Herold-Mende, A. Benner, M. Thome, N. Gassler, W. Wahl, S. Lyer, A. Poustka, H. F. Otto, M. Deichmann, BRAF mutations distinguish anorectal from cutaneous melanoma at the molecular level. Gastroenterology 127, 1815–1820 (2004).
44
I. Satzger, T. Schaefer, U. Kuettler, V. Broecker, B. Voelker, H. Ostertag, A. Kapp, R. Gutzmer, Analysis of c-KIT expression and KIT gene mutation in human mucosal melanomas. Br. J. Cancer 99, 2065–2069 (2008).
45
H.-Y. Li, L. Zhou, J. Tian, N. Cong, [Analysis of BRAF mutation and extracellular regulated protein kinases activation in nasal mucosa melanomas]. Zhonghua Yi Xue Za Zhi 90, 3399–3402 (2010).
46
J. Yun, J. Lee, J. Jang, E. J. Lee, K. T. Jang, J. H. Kim, K.-M. Kim, KIT amplification and gene mutations in acral/mucosal melanoma in Korea. APMIS 119, 330–335 (2011).
47
Y. Cohen, N. Goldenberg-Cohen, S. Akrish, T. Shani, N. Amariglio, O. Dratviman-Storobinsky, I. Kaplan, I. Barshack, A. Hirshberg, BRAF and GNAQ mutations in melanocytic tumors of the oral cavity. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. 114, 778–784 (2012).
48
S. Ni, D. Huang, X. Chen, J. Huang, Y. Kong, Y. Xu, X. Du, W. Sheng, c-kit gene mutation and CD117 expression in human anorectal melanomas. Hum. Pathol. 43, 801–807 (2012).
49
M. Turri-Zanoni, D. Medicina, D. Lombardi, M. Ungari, P. Balzarini, C. Rossini, W. Pellegrini, P. Battaglia, C. Capella, P. Castelnuovo, G. Palmedo, F. Facchetti, H. Kutzner, P. Nicolai, W. Vermi, Sinonasal mucosal melanoma: Molecular profile and therapeutic implications from a series of 32 cases. Head Neck 35, 1066–1077 (2013).
50
W. O. Greaves, S. Verma, K. P. Patel, M. A. Davies, B. A. Barkoh, J. M. Galbincea, H. Yao, A. J. Lazar, K. D. Aldape, L. J. Medeiros, R. Luthra, Frequency and spectrum of BRAF mutations in a retrospective, single-institution study of 1112 cases of melanoma. J. Mol. Diagn. 15, 220–226 (2013).
51
A. Zebary, M. Jangard, K. Omholt, B. Ragnarsson-Olding, J. Hansson, KIT, NRAS and BRAF mutations in sinonasal mucosal melanoma: A study of 56 cases. Br. J. Cancer 109, 559–564 (2013).
52
M. Chraybi, I. Abd Alsamad, C. Copie-Bergman, M. Baia, J. André, N. Dumaz, N. Ortonne, Oncogene abnormalities in a series of primary melanomas of the sinonasal tract: NRAS mutations and cyclin D1 amplification are more frequent than KIT or BRAF mutations. Hum. Pathol. 44, 1902–1911 (2013).
53
M. Colombino, A. Lissia, R. Franco, G. Botti, P. A. Ascierto, A. Manca, M. C. Sini, M. Pisano, P. Paliogiannis, F. Tanda, G. Palmieri, A. Cossu, Unexpected distribution of cKIT and BRAF mutations among southern Italian patients with sinonasal melanoma. Dermatology 226, 279–284 (2013).
54
S. Aulmann, H. P. Sinn, R. Penzel, C. B. Gilks, S. Schott, J. C. Hassel, D. Schmidt, F. Kommoss, P. Schirmacher, S. Kommoss, Comparison of molecular abnormalities in vulvar and vaginal melanomas. Mod. Pathol. 27, 1386–1393 (2014).
55
A. C. H. van Engen-van Grunsven, H. V. N. Küsters-Vandevelde, J. De Hullu, L. M. van Duijn, J. Rijntjes, J. V. M. G. Bovée, P. J. T. A. Groenen, W. A. M. Blokx, NRAS mutations are more prevalent than KIT mutations in melanoma of the female urogenital tract—A study of 24 cases from the Netherlands. Gynecol. Oncol. 134, 10–14 (2014).
56
R. Santi, L. Simi, R. Fucci, M. Paglierani, M. Pepi, P. Pinzani, B. Merelli, M. Santucci, G. Botti, C. Urso, D. Massi, KIT genetic alterations in anorectal melanomas. J. Clin. Pathol. 68, 130–134 (2015).
57
D. Tseng, J. Kim, A. Warrick, D. Nelson, M. Pukay, C. Beadling, M. Heinrich, M. A. Selim, C. L. Corless, K. Nelson, Oncogenic mutations in melanomas and benign melanocytic nevi of the female genital tract. J. Am. Acad. Dermatol. 71, 229–236 (2014).
58
K. I. Pappa, G. D. Vlachos, M. Roubelakis, D.-E. G. Vlachos, T. G. Kalafati, D. Loutradis, N. P. Anagnou, Low mutational burden of eight genes involved in the MAPK/ERK, PI3K/AKT, and GNAQ/11 pathways in female genital tract primary melanomas. BioMed Res. Int. 2015, 303791 (2015).
59
M. Rouzbahman, S. Kamel-Reid, A. Al Habeeb, M. Butler, J. Dodge, S. Laframboise, J. Murphy, G. Rasty, D. Ghazarian, Malignant melanoma of vulva and vagina: A histomorphological review and mutation analysis—A single-center study. J. Low. Genit. Tract Dis. 19, 350–353 (2015).
60
M. Jangard, A. Zebary, B. Ragnarsson-Olding, J. Hansson, TERT promoter mutations in sinonasal malignant melanoma: A study of 49 cases. Melanoma Res. 25, 185–188 (2015).
61
H. M. Yang, S. J. Hsiao, D. F. Schaeffer, C. Lai, H. E. Remotti, D. Horst, M. M. Mansukhani, B. A. Horst, Identification of recurrent mutational events in anorectal melanoma. Mod. Pathol. 30, 286–296 (2017).
62
O. Yélamos, E. A. Merkel, L. M. Sholl, B. Zhang, S. M. Amin, C. Y. Lee, G. E. Guitart, J. Yang, A. T. Wenzel, C. G. Bunick, P. Yazdan, J. Choi, P. Gerami, Nonoverlapping clinical and mutational patterns in melanomas from the female genital tract and atypical genital nevi. J. Invest. Dermatol. 136, 1858–1865 (2016).
63
J. Lyu, Y. Wu, C. Li, R. Wang, H. Song, G. Ren, W. Guo, Mutation scanning of BRAF, NRAS, KIT, and GNAQ/GNA11 in oral mucosal melanoma: A study of 57 cases. J. Oral Pathol. Med. 45, 295–301 (2016).
64
X. Sheng, Y. Kong, Y. Li, Q. Zhang, L. Si, C. Cui, Z. Chi, B. Tang, L. Mao, B. Lian, X. Wang, X. Yan, S. Li, J. Dai, J. Guo, GNAQ and GNA11 mutations occur in 9.5% of mucosal melanoma and are associated with poor prognosis. Eur. J. Cancer 65, 156–163 (2016).
65
J. D. Hintzsche, N. T. Gorden, C. M. Amato, J. Kim, K. E. Wuensch, S. E. Robinson, A. J. Applegate, K. L. Couts, T. M. Medina, K. R. Wells, J. A. Wisell, M. D. McCarter, N. F. Box, Y. G. Shellman, R. C. Gonzalez, K. D. Lewis, J. J. Tentler, A. C. Tan, W. A. Robinson, Whole-exome sequencing identifies recurrent SF3B1 R625 mutation and comutation of NF1 and KIT in mucosal melanoma. Melanoma Res. 27, 189–199 (2017).
66
J. Y. Hou, C. Baptiste, R. B. Hombalegowda, A. I. Tergas, R. Feldman, N. L. Jones, S. Chatterjee-Paer, A. Bus-Kwolfski, J. D. Wright, W. M. Burke, Vulvar and vaginal melanoma: A unique subclass of mucosal melanoma based on a comprehensive molecular analysis of 51 cases compared with 2253 cases of nongynecologic melanoma. Cancer 123, 1333–1344 (2017).
67
N. K. Hayward, J. S. Wilmott, N. Waddell, P. A. Johansson, M. A. Field, K. Nones, A.-M. Patch, H. Kakavand, L. B. Alexandrov, H. Burke, V. Jakrot, S. Kazakoff, O. Holmes, C. Leonard, R. Sabarinathan, L. Mularoni, S. Wood, Q. Xu, N. Waddell, V. Tembe, G. M. Pupo, R. De Paoli-Iseppi, R. E. Vilain, P. Shang, L. M. S. Lau, R. A. Dagg, S.-J. Schramm, A. Pritchard, K. Dutton-Regester, F. Newell, A. Fitzgerald, C. A. Shang, S. M. Grimmond, H. A. Pickett, J. Y. Yang, J. R. Stretch, A. Behren, R. F. Kefford, P. Hersey, G. V. Long, J. Cebon, M. Shackleton, A. J. Spillane, R. P. M. Saw, N. López-Bigas, J. V. Pearson, J. F. Thompson, R. A. Scolyer, G. J. Mann, Whole-genome landscapes of major melanoma subtypes. Nature 545, 175–180 (2017).
68
Ş. Öztürk Sari, İ. Yilmaz, O. Ç. Taşkin, G. Narli, F. Şen, Ş. Çomoğlu, P. Firat, B. Bİlgİç, D. Yilmazbayhan, Y. Özlük, N. Büyükbabanİ, BRAF, NRAS, KIT, TERT, GNAQ/GNA11 mutation profile analysis of head and neck mucosal melanomas: A study of 42 cases. Pathology 49, 55–61 (2017).
69
I. Cosgarea, S. Ugurel, A. Sucker, E. Livingstone, L. Zimmer, M. Ziemer, J. Utikal, P. Mohr, C. Pfeiffer, C. Pföhler, U. Hillen, S. Horn, D. Schadendorf, K. G. Griewank, A. Roesch, Targeted next generation sequencing of mucosal melanomas identifies frequent NF1 and RAS mutations. Oncotarget 8, 40683–40692 (2017).
70
J. Lyu, Z. Song, J. Chen, M. J. Shepard, H. Song, G. Ren, Z. Li, W. Guo, Z. Zhuang, Y. Shi, Whole-exome sequencing of oral mucosal melanoma reveals mutational profile and therapeutic targets. J. Pathol. 244, 358–366 (2018).
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This is an article distributed under the terms of the Science Journals Default License.
History
Received: 12 July 2018
Accepted: 11 October 2018
1 November 2018
Acknowledgments
We thank C. Onodera and E. Talevich for helpful advice during data analysis. Many thanks to S. Liu and E. Martin for excellent technical assistance. We also thank S. Avagyan and M. Fazio for critical reading of the manuscript and helpful discussions. Funding: Funding for this study was provided by the Melanoma Research Foundation, the Dermatology Foundation, the Melanoma Research Alliance, the Ellison Foundation, the Starr Foundation, the V Foundation, the Terry Patters Melanoma Foundation, and the National Cancer Institute (1R35CA220481 to B.C.B., R01CA103846 to L.I.Z., K99CA201465 to J.A., U24CA196067 to R.C.J., and T32HL007627 to J.K.M.). L.I.Z. is a Howard Hughes Medical Institute Investigator. Author contributions: I.Y., J.A., M.X., B.C.B., and L.I.Z. designed the study. I.Y., B.C.B., H.W., N.M.J., B.H.D., and R.C.J. performed sample selection and collection. M.X. and I.Y. performed genetic analyses. M.X. and I.Y. performed immunohistochemistry, and I.Y. analyzed the results. J.A., H.R., and C.F.B. performed zebrafish experiments, and J.A. and J.K.M. analyzed the results. J.A. performed human melanoma cell line experiments and analyzed the results. I.Y., J.A., M.X., B.C.B., and L.I.Z. wrote the manuscript, and all authors provided constructive feedback. Competing interests: I.Y. received grants from Ignyta, Novartis, and Bayer, outside the submitted work. B.C.B. is a consultant for Lilly, Inc., outside the submitted work. L.I.Z. is a founder and stockholder of Fate Therapeutics, CAMP4 Therapeutics, and Scholar Rock, outside the submitted work. All other authors declare no conflicts of interest. Data and materials availability: Mucosal melanoma targeted DNA sequencing data have been deposited in dbGAP (project phs001594.v1). All plasmids described in this study have been deposited at Addgene.
Authors
Funding Information
National Institutes of Health: R01CA103846
National Cancer Institute: 1R35CA220481
National Cancer Institute: K99CA201465
National Cancer Institute: U24CA196067
Terry Patters Melanoma Foundation:
The Ellison Foundation:
