Inherited Genetic Susceptibility to Nonimmunosuppressed Epstein-Barr Virus-associated T/NK-cell Lymphoproliferative Diseases in Chinese Patients

Abstract

Epstein-Barr virus (EBV) T/NK-cell lymphoproliferative diseases are characterized by clonal expansion of EBV-infected T or NK cells, including chronic active EBV infection of T/NK-cell type (CAEBV⁺T/NK), EBV-associated hemophagocytic lymphohistiocytosis (EBV⁺HLH), extranodal NK/T-cell lymphoma of nasal type (ENKTL), and aggressive NK-cell leukemia (ANKL). However, the role of inherited genetic variants to EBV⁺T/NK-LPDs susceptibility is still unknown. A total of 171 nonimmunosuppressed patients with EBV⁺T/NK-LPDs and 104 healthy donors were retrospectively collected and a targeted sequencing study covering 15 genes associated with lymphocyte cytotoxicity was performed. The 94 gene variants, mostly located in UNC13D, LYST, ITK, and PRF1 genes were detected, and mutations covered 28/50 (56.00%) of CAEBV-T/NK, 31/51 (60.78%) of EBV⁺HLH, 13/28 (46.42%) of ENKTL, and 13/48 (27.09%) of ANKL. Most mutations represented monoallelic and missense. Three-year overall survival rate of patients with CAEBV-T/NK and EBV⁺HLH was significantly lower in patients with germline mutations than in those without germline mutations (P=0.0284, P=0.0137). Our study provided novel insights into understanding a spectrum of nonimmunosuppressed EBV⁺T/NK-LPDs with respect to genetic defects associated with lymphocyte cytotoxicity and reminded us that the gene sequencing may be an auxiliary test for diagnosis and risk stratification of EBV⁺T/NK-LPDs.

Keywords

germline mutation / EBV-associated T/NK-cell lymphoproliferative disease / hemophagocytic lymphohistiocytosis / primary immunodeficiencies / lymphocyte cytotoxicity / gene sequencing

References

[1] DelecluseHJ, FeederleR, O’SullivanB, et al.. Epstein Barr virus-associated tumours: an update for the attention of the working pathologist. J Clin Pathol, 2007, 60(12): 1358-1364

[2] TannerJ, WeisJ, FearonD, et al.. Epstein-barr virus gp350/220 binding to the B lymphocyte C3d receptor mediates adsorption, capping, and endocytosis. Cell, 1987, 50(2): 203-213

[3] KimuraH, ItoY, KawabeS, et al.. EBV-associated T/ NK-cell lymphoproliferative diseases in nonimmuno-compromised hosts: prospective analysis of 108 cases. Blood, 2012, 119(3): 673-686

[4] HenterJI, HorneA, AricóM, et al.. HLH-2004: Diagnostic and therapeutic guidelines for hemophagocytic lymphohistiocytosis. Pediat Blood Cancer, 2007, 48(2): 124-131

[5] ArberDA, OraziA, HasserjianR, et al.. The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood, 2016, 127(20): 2391-2405

[6] SongSY, KimWS, KoYH, et al.. Aggressive natural killer cell leukemia: clinical features and treatment outcome. Haematologica, 2002, 87(12): 1343-1345

[7] OhshimaK, KimuraH, YoshinoT, et al.. Proposed categorization of pathological states of EBV-associated T/natural killer-cell lymphoproliferative disorder (LPD) in children and young adults: overlap with chronic active EBV infection and infantile fulminant EBV T-LPD. Pathol Int, 2008, 58(4): 209-217

[8] KimuraH, KawadaJ, ItoY. Epstein-Barr virus-associated lymphoid malignancies: the expanding spectrum of hematopoietic neoplasms. Nagoya J Med Sci, 2013, 75(3–4): 169-179

[9] SteppSE, Dufourcq-LagelouseR, DeistFL, et al.. Perforin gene defects in familial hemophagocytic lymphohistiocytosis. Science, 1999, 286(5446): 1957-1959

[10] FeldmannJ, CallebautI, RaposoG, et al.. Munc13–4 is essential for cytolytic granules fusion and is mutated in a form of familial hemophagocytic lymphohistiocytosis (FHL3). Cell, 2003, 115(4): 461-473

[11] zur StadtU, RohrJ, SeifertW, et al.. Familial hemophagocytic lymphohistiocytosis type 5 (FHL-5) is caused by mutations in Munc18–2 and impaired binding to syntaxin 11. Am J Hum Genet, 2009, 85(4): 482-492

[12] ArnesonLN, BrickshawanaA, SegovisCM, et al.. Cutting edge: syntaxin 11 regulates lymphocyte-mediated secretion and cytotoxicity. J Immunol, 2007, 179(6): 3397-3401

[13] BizarioJC, FeldmannJ, CastroFA, et al.. Griscelli syndrome: characterization of a new mutation and rescue of T-cytotoxic activity by retroviral transfer of RAB27A gene. J Clin Immunol, 2004, 24(4): 397-410

[14] RigaudS, FondanècheMC, LambertN, et al.. XIAP deficiency in humans causes an X-linked lymphoproliferative syndrome. Nature, 2006, 444(7115): 110-114

[15] AricoM, ImashukuS, ClementiR, et al.. Hemophagocytic lymphohistiocytosis due to germline mutations in SH2D1A, the X-linked lymphoproliferative disease gene. Blood, 2001, 97(4): 1131-1133

[16] HeibeinJA, GopingIS, BarryM, et al.. Granzyme B-mediated cytochrome c release is regulated by the Bcl-2 family members bid and Bax. J Exp Med, 2000, 192(10): 1391-1402

[17] WardDM, ShiflettSL, KaplanJ. Chediak-Higashi syndrome: a clinical and molecular view of a rare lysosomal storage disorder. Curr Mol Med, 2002, 2(5): 469-477

[18] TangyeSG, PalendiraU, EdwardsES. Human immunity against EBV-lessons from the clinic. J Exp Med, 2017, 214(2): 269-283

[19] KimuraH, MorishimaT, KaneganeH, et al.. Prognostic factors for chronic active Epstein-Barr virus infection. J Infect Dis, 2003, 187(4): 527-533

[20] OkanoM, KawaK, KimuraH, et al.. Proposed guidelines for diagnosing chronic active Epstein-Barr virus infection. Am J Hematol, 2005, 80(1): 64-69

[21] MiaoY, ZhuHY, QiaoC, et al.. Pathogenic gene mutations or variants identified by targeted gene sequencing in adults with hemophagocytic lymphohistiocytosis. Front Immunol, 2019, 10: 395-395

[22] VoskoboinikI, ThiaMC, TrapaniJA. A functional analysis of the putative polymorphisms A91V and N252S and 22 missense perforin mutations associated with familial hemophagocytic lymphohistiocytosis. Blood, 2005, 105(12): 4700-4706

[23] YoonHS, KimHJ, YooKH, et al.. UNC13D is the predominant causative gene with recurrent splicing mutations in Korean patients with familial hemophagocytic lymphohistiocytosis. Haematologica, 2010, 95(6): 622-626

[24] CoteM, MenagerMM, BurgessA, et al.. Munc18–2 deficiency causes familial hemophagocytic lymphohistiocytosis type 5 and impairs cytotoxic granule exocytosis in patient NK cells. J Clin Invest, 2009, 119(12): 3765-3773

[25] Zur StadtU, SchmidtS, KasperB, et al.. Linkage of familial hemophagocytic lymphohistiocytosis (FHL) type-4 to chromosome 6q24 and identification of mutations in syntaxin 11. Hum Mol Genet, 2005, 14(6): 827-834

[26] ZhangKJ, JordanMB, MarshRA, et al.. Hypomorphic mutations in PRF1, MUNC13–4, and STXBP2 are associated with adult-onset familial HLH. Blood, 2011, 118(22): 5794-5798

[27] ZhangKJ, ChandrakasanS, ChapmanH, et al.. Synergistic defects of different molecules in the cytotoxic pathway lead to clinical familial hemophagocytic lymphohistiocytosis. Blood, 2014, 124(8): 1331-1334

[28] MeethsM, BrycesonYT. HLH susceptibility: genetic lesions add up. Blood, 2016, 127(17): 2051-2052

[29] SepulvedaFE, GarrigueA, MaschalidiS, et al.. Polygenic mutations in the cytotoxicity pathway increase susceptibility to develop HLH immunopathology in mice. Blood, 2016, 127(17): 2113-2121

[30] LöfstedtA, ChiangSC, OnelövE, et al.. Cancer risk in relatives of patients with a primary disorder of lymphocyte cytotoxicity: a retrospective cohort study. Lancet Haematol, 2015, 2(12): e536-542

[31] ChiaJ, YeoKP, WhisstockJC, et al.. Temperature sensitivity of human perforin mutants unmasks subtotal loss of cytotoxicity, delayed FHL, and a predisposition to cancer. Proc Natl Acad Sci USA, 2009, 106(24): 9809-9814

[32] MansoR, Rodriguez-PinillaSM, LombardiaL, et al.. An A91V SNP in the perforin gene is frequently found in NK/T-cell lymphomas. PLoS One, 2014, 9(3): e91521

[33] CohenJI, NiemelaJE, StoddardJL, et al.. Late-onset severe chronic active EBV in a patient for five years with mutations in STXBP2 (MUNC18–2) and PRF1 (perforin 1). J Clin Immunol, 2015, 35(5): 445-448

[34] KatanoH, AliMA, PateraAC, et al.. Chronic active Epstein-Barr virus infection associated with mutations in perforin that impair its maturation. Blood, 2004, 103(4): 1244-1252

[35] TongCR, LiuHX, XieJJ, et al.. The study of gene mutations in unknown refractory viral infection and primary hemophagocytic lymphohistiocytosis. Zhonghua Nei Ke Za Zhi (Chinese), 2011, 50(4): 280-283

[36] RichardsS, AzizN, BaleS, et al.. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med, 2015, 17(5): 405-424