Areca nut (AN) chewing contributes to an increase of oral squamous cell carcinoma (OSCC) cases in South and Southeast Asia; however, genomic events underlying the carcinogenesis process of AN-related OSCC remain unclear. Here, we comprehensively describe the genomic and transcriptome alterations of 113 Chinese OSCC patients (89 AN related and 24 AN negative) by whole-exome sequencing and RNA sequencing, and we compared the genomic differences between AN-related and AN-negative samples by integrating sequencing data of 325 OSCC patients from The Cancer Genome Atlas database and 50 from a published Taiwanese study. We identified 11 significantly mutated genes for OSCC, including 4 novel ones (ATG2A, WEE1, DST, and TSC2), of which WEE1 and ATG2A mutated with significantly higher rates in AN-related samples (P = 0.04 and P = 0.003, respectively). Mutational signature analysis revealed that AN-related OSCCs were specially characterized by the genomic signature of mismatch repair deficiency (dMMR), which could also predict the prognosis status of AN-related OSCC. In addition, an elevated PD-L1 expression was also observed in both AN-related patients (P = 3.71 × 10−11) and those with a high dMMR level (P = 1.99 × 10−4). Further differential expression analysis and in vitro experiments confirmed the role of dMMR in the development of OSCC induced by AN exposure. Taken together, this study first revealed the molecular profiles and highlighted the role of dMMR in AN-related OSCC among the Chinese population and identified that AN-related OSCC may represent a potential cohort for effective anti-PD-1/L1 immunotherapy.
AgrawalNFrederickMJPickeringCRBettegowdaCChangKLiRJFakhryCXieTXZhangJWangJ, et al. 2011. Exome sequencing of head and neck squamous cell carcinoma reveals inactivating mutations in NOTCH1. Science. 333(6046):1154–1157.
2.
BridgeGRashidSMartinSA. 2014. DNA mismatch repair and oxidative DNA damage: implications for cancer biology and treatment. Cancers. 6(3):1597–1614.
3.
CalveteOGarcia-PaviaPDomínguezFBougeardGKunzeKBraeuningerATeuleALasaARamónYCajalTLlortG, et al. 2017. The wide spectrum of POT1 gene variants correlates with multiple cancer types. Eur J Hum Genet. 25(11):1278–1281.
4.
ChenTWLeeCCLiuHWuCSPickeringCRHuangPJWangJChangIYYehYMChenCD, et al. 2017. APOBEC3A is an oral cancer prognostic biomarker in Taiwanese carriers of an APOBEC deletion polymorphism. Nat Commun. 8(1):465.
5.
ChenYLLiuKJJangCWHsuCCYenYCLiuYLChuangTHWangSHFuYKKuoCC, et al. 2019. ERK activation modulates cancer stemness and motility of a novel mouse oral squamous cell carcinoma cell line. Cancers. 12(1). pii: E61.
6.
ColussiCParlantiEDeganPAquilinaGBarnesDMacphersonPKarranPCrescenziMDogliottiEBignamiM.2002. The mammalian mismatch repair pathway removes DNA 8-oxodgmp incorporated from the oxidized dNTP pool. Curr Biol. 12(11):912–918.
7.
CzerninskiRKrichevskySAshhabYGazitDPatelVBen-YehudaD.2009. Promoter hypermethylation of mismatch repair genes, hMLH1 and hMSH2 in oral squamous cell carcinoma. Oral Dis. 15(3):206–213.
8.
DoKDoroshowJHKummarS.2013. Wee1 kinase as a target for cancer therapy. Cell Cycle. 12(19):3159–3164.
9.
Fernández-MateosJSeijas-TamayoRKlainJCABorgoñónMPPérez-RuizEMesíaRDel BarcoEColomaCSDominguezARDaroquiJC, et al. 2017. Analysis of autophagy gene polymorphisms in Spanish patients with head and neck squamous cell carcinoma. Sci Rep. 7(1):6887.
10.
FerrisRLBlumenscheinGJrFayetteJGuigayJColevasADLicitraLHarringtonKKasperSVokesEEEvenC, et al. 2016. Nivolumab for recurrent squamous-cell carcinoma of the head and neck. N Engl J Med. 375(19):1856–1867.
11.
GadhikarMAZhangJShenLRaoXWangJZhaoMKaluNNJohnsonFMByersLAHeymachJ, et al. 2018. CDKN2A/p16 deletion in head and neck cancer cells is associated with CDK2 activation, replication stress, and vulnerability to CHK1 inhibition. Cancer Res. 78(3):781–797.
12.
Gomez-SanchezRRoseJGuimaraesRMariMPapinskiDRieterEGeertsWJHardenbergRKraftCUngermannC, et al. 2018. Atg9 establishes Atg2-dependent contact sites between the endoplasmic reticulum and phagophores. J Cell Biol. 217(8):2743–2763.
13.
HuYJChenJZhongWSLingTYJianXCLuRHTangZGTaoL.2017. Trend analysis of betel nut-associated oral cancer and health burden in China. Chin J Dent Res. 20(2):69–78.
14.
IlleperumaRPKimDKParkYJSonHKKimJYKimJLeeDYKimKYJungDWTilakaratneWM, et al. 2015. Areca nut exposure increases secretion of tumor-promoting cytokines in gingival fibroblasts that trigger DNA damage in oral keratinocytes. Int J Cancer. 137(11):2545–2557.
15.
JengJHChangMCHahnLJ. 2001. Role of areca nut in betel quid-associated chemical carcinogenesis: current awareness and future perspectives. Oral Oncol. 37(6):477–492.
16.
KhanIPantINarraSRadheshRRanganathanKRaoSGKondaiahP.2015. Epithelial atrophy in oral submucous fibrosis is mediated by copper (ii) and arecoline of areca nut. J Cell Mol Med. 19(10):2397–2412.
17.
LeeVLeDT. 2016. Efficacy of PD-1 blockade in tumors with MMR deficiency. Immunotherapy. 8(1):1–3.
18.
LiYCChangJTChiuCLuYCLiYLChiangCHYouGRLeeLYChengAJ. 2016. Areca nut contributes to oral malignancy through facilitating the conversion of cancer stem cells. Mol Carcinog. 55(5):1012–1023.
19.
MacphersonPBaroneFMagaGMazzeiFKarranPBignamiM.2005. 8-Oxoguanine incorporation into DNA repeats in vitro and mismatch recognition by mutsalpha. Nucleic Acids Res. 33(16):5094–5105.
20.
McKayJDHungRJHanYZongXCarreras-TorresRChristianiDCCaporasoNEJohanssonMXiaoXLiY, et al. 2017. Large-scale association analysis identifies new lung cancer susceptibility loci and heterogeneity in genetic susceptibility across histological subtypes. Nat Genet. 49(7):1126–1132.
21.
MortazaviHBaharvandMMehdipourM.2014. Oral potentially malignant disorders: an overview of more than 20 entities. J Dent Res Dent Clin Dent Prospects. 8(1):6–14.
22.
MusicDDahlrotRHHermansenSKHjelmborgJde StrickerKHansenSKristensenBW. 2016. Expression and prognostic value of the WEE1 kinase in gliomas. J Neurooncol. 127(2):381–389.
23.
NojimaHHommaHOnozatoYKaidaAHaradaHMiuraM.2020. Differential properties of mitosis-associated events following CHK1 and WEE1 inhibitor treatments in human tongue carcinoma cells. Exp Cell Res. 386(2):111720.
24.
ParryDPetersG.1996. Temperature-sensitive mutants of p16CDKN2 associated with familial melanoma. Mol Cell Biol. 16(7):3844–3852.
25.
PickeringCRZhangJYooSYBengtssonLMoorthySNeskeyDMZhaoMOrtega AlvesMVChangKDrummondJ, et al. 2013. Integrative genomic characterization of oral squamous cell carcinoma identifies frequent somatic drivers. Cancer Discov. 3(7):770–781.
26.
PrabhuRVPrabhuVChatraLShenaiPSuvarnaNDandekeriS.2014. Areca nut and its role in oral submucous fibrosis. J Clin Exp Dent. 6(5):e569–e575.
27.
ShieldKDFerlayJJemalASankaranarayananRChaturvediAKBrayFSoerjomataramI.2017. The global incidence of lip, oral cavity, and pharyngeal cancers by subsite in 2012. CA Cancer J Clin. 67(1):51–64.
28.
ShinKHParkKHHongHJKimJMOhJEChoungPHMinBM. 2002. Prevalence of microsatellite instability, inactivation of mismatch repair genes, p53 mutation, and human papillomavirus infection in Korean oral cancer patients. Int J Oncol. 21(2):297–302.
29.
SuSCLinCWLiuYFFanWLChenMKYuCPYangWESuCWChuangCYLiWH, et al. 2017. Exome sequencing of oral squamous cell carcinoma reveals molecular subgroups and novel therapeutic opportunities. Theranostics. 7(5):1088–1099.
30.
SunLMooreEBermanRClavijoPESalehAChenZVan WaesCDaviesJFriedmanJAllenCT. 2018. WEE1 kinase inhibition reverses G2/M cell cycle checkpoint activation to sensitize cancer cells to immunotherapy. Oncoimmunology. 7(10):e1488359.
31.
TangZTakahashiYChenCLiuYHeHTsotakosNSerfassJMGebruMTChenHYoungMM, et al. 2017. Atg2A/B deficiency switches cytoprotective autophagy to non-canonical caspase-8 activation and apoptosis. Cell Death Differ. 24(12):2127–2138.
32.
TsaiYSLeeKWHuangJLLiuYSJuoSHKuoWRChangJGLinCSJongYJ. 2008. Arecoline, a major alkaloid of areca nut, inhibits p53, represses DNA repair, and triggers DNA damage response in human epithelial cells. Toxicology. 249(2–3):230–237.
33.
van HartenAMBuijzeMvan der MastRRooimansMAMartens-de KempSRBachasCBrinkAStigter-van WalsumMWolthuisRMFBrakenhoffRH. 2019. Targeting the cell cycle in head and neck cancer by Chk1 inhibition: a novel concept of bimodal cell death. Oncogenesis. 8(7):38.
34.
VelikkakathAKNishimuraTOitaEIshiharaNMizushimaN.2012. Mammalian Atg2 proteins are essential for autophagosome formation and important for regulation of size and distribution of lipid droplets. Mol Biol Cell. 23(5):896–909.
35.
VriendLEDe Witt HamerPCVan NoordenCJWurdingerT.2013. WEE1 inhibition and genomic instability in cancer. Biochim Biophys Acta. 1836(2):227–235.
36.
WangTYPengCYLeeSSChouMYYuCCChangYC. 2016. Acquisition cancer stemness, mesenchymal transdifferentiation, and chemoresistance properties by chronic exposure of oral epithelial cells to arecoline. Oncotarget. 7(51):84072–84081.
37.
WangYZhouXSongYJiXZhangAZhangGGaoZ.2013. The mismatch repair gene hPMS1 (human postmeiotic segregation1) is down regulated in oral squamous cell carcinoma. Gene. 524(1):28–34.
38.
WuFParvezFIslamTAhmedARakibuz-ZamanMHasanRArgosMLevyDSarwarGAhsanH, et al. 2015. Betel quid use and mortality in Bangladesh: a cohort study. Bull World Health Organ. 93(10): 684–692.
39.
YangSHLeeTYHoCAYangCYHuangWYLinYCNiehSLinYSChenSFLinFH. 2018. Exposure to nicotine-derived nitrosamine ketone and arecoline synergistically facilitates tumor aggressiveness via overexpression of epidermal growth factor receptor and its downstream signaling in head and neck squamous cell carcinoma. PLoS One. 13(8):e0201267.
40.
YenCYChiangWFLiuSYChengPCLeeSYHongWZLinPYLinMHLiuYC. 2014. Long-term stimulation of areca nut components results in increased chemoresistance through elevated autophagic activity. J Oral Pathol Med. 43(2):91–96.
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