Mutations in JAK2, MPL and CALR genes have been identified in the majority of myeloproliferative neoplasm (MPN) patients, and patients negative for these three mutations are the so-called triple-negative (TN) MPN. In this study, we examined the mutational profiles of 16 triple-negative MPN patients including 7 essential thrombocythemia (ET), 1 primary myelofibrosis and 8 polycythemia vera (PV). Targeted next-generation sequencing was performed using the ACTOnco Comprehensive Cancer Panel (Ion AmpliSeq Comprehensive Cancer Panel, Life Technologies) to target all coding exons of 409 cancer-related genes. Overall, 30 nonsynonymous somatic mutations were detected in 12 (75%) patients with a range of 1–5 mutations per sample. Notably, one ET patient was found to have JAK2V617F and KITP551L mutations at very low allele frequency. One MPLP70L and 1 MPLM602T mutations were identified each in 1 ET and 1 PV, respectively. Other recurrent mutations were also identified including KMT2C, KMT2D, IRS2, SYNE1, PDE4DIP, SETD2, ATM, TNFAIP3 and CCND2. In addition, germline mutations were also found in some cancer-related genes. Copy number changes were rare in this cohort of TN MPNs. In conclusion, both somatic and germline mutations can be detected in TN MPN patients.
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We are grateful to Drs. Kuei-Fang Chou, Po-Nien Liao and Guan-Jhe Cai for their help in patient enrollment.
This study was supported by Grants from the Ministry of Science and Technology of Taiwan to KHL (Grant Numbers: MOST 102-2314-B-195-015-MY2 and MOST 104-2314-B-195-017-MY3), and the intramural Grants from the Department of Medical Research of MacKay Memorial Hospital to KHL and YCC. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Compliance with ethical standards
Conflict of interest
Dr. Shu-Jen Chen is an employee of ACTGenomics, Co. Ltd. All the other authors declare that they have no conflict of interest.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
James C, Ugo V, Le Couedic JP, Staerk J, Delhommeau F, Lacout C, et al. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005;434(7037):1144–8.CrossRefPubMedGoogle Scholar
Baxter EJ, Scott LM, Campbell PJ, East C, Fourouclas N, Swanton S, et al. Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. Lancet. 2005;365(9464):1054–61.CrossRefPubMedGoogle Scholar
Kralovics R, Passamonti F, Buser AS, Teo SS, Tiedt R, Passweg JR, et al. A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med. 2005;352(17):1779–90.CrossRefPubMedGoogle Scholar
Levine RL, Wadleigh M, Cools J, Ebert BL, Wernig G, Huntly BJ, et al. Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. Cancer Cell. 2005;7(4):387–97.CrossRefPubMedGoogle Scholar
Klampfl T, Gisslinger H, Harutyunyan AS, Nivarthi H, Rumi E, Milosevic JD, et al. Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med. 2013;369(25):2379–90.CrossRefPubMedGoogle Scholar
Nangalia J, Massie CE, Baxter EJ, Nice FL, Gundem G, Wedge DC, et al. Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2. N Engl J Med. 2013;369(25):2391–405.CrossRefPubMedPubMedCentralGoogle Scholar
Lim K-H, Lin H-C, Chen CG-S, Chiang Y-H, Hsiao C-D, Kuo Y-Y. CALR mutations in myeloproliferative neoplasms. Int J Gerontol. 2014;8(2):105.CrossRefGoogle Scholar
Kilpivaara O, Levine RL. JAK2 and MPL mutations in myeloproliferative neoplasms: discovery and science. Leukemia. 2008;22(10):1813–7.CrossRefPubMedGoogle Scholar
Pikman Y, Lee BH, Mercher T, McDowell E, Ebert BL, Gozo M, et al. MPLW515L is a novel somatic activating mutation in myelofibrosis with myeloid metaplasia. PLoS Med. 2006;3(7):e270.CrossRefPubMedPubMedCentralGoogle Scholar
Tefferi A. Novel mutations and their functional and clinical relevance in myeloproliferative neoplasms: JAK2, MPL, TET2, ASXL1, CBL, IDH and IKZF1. Leukemia. 2010;24(6):1128–38.CrossRefPubMedPubMedCentralGoogle Scholar
Pardanani A, Lasho T, Finke C, Oh ST, Gotlib J, Tefferi A. LNK mutation studies in blast-phase myeloproliferative neoplasms, and in chronic-phase disease with TET2, IDH, JAK2 or MPL mutations. Leukemia. 2010;24(10):1713–8.CrossRefPubMedGoogle Scholar
Lin H-C, Wang S-C, Chen CG-S, Chang M-C, Wang W-T, Su N-W, et al. Mutation and lineage analysis of DNMT3A in BCR-ABL1-negative chronic myeloproliferative neoplasms. Int J Gerontol. 2013;7(3):186–8.CrossRefGoogle Scholar
Milosevic Feenstra JD, Nivarthi H, Gisslinger H, Leroy E, Rumi E, Chachoua I, et al. Whole-exome sequencing identifies novel MPL and JAK2 mutations in triple-negative myeloproliferative neoplasms. Blood. 2016;127(3):325–32.CrossRefPubMedPubMedCentralGoogle Scholar
Cabagnols X, Favale F, Pasquier F, Messaoudi K, Defour JP, Ianotto JC, et al. Presence of atypical thrombopoietin receptor (MPL) mutations in triple-negative essential thrombocythemia patients. Blood. 2016;127(3):333–42.CrossRefPubMedGoogle Scholar
Lin H-C, Chen CG-S, Chang M-C, Wang W-T, Kao CW, Lo A-C, et al. JAK2 V617F mutation in adult Taiwanese patients with essential thrombocythemia: more prevalent in old patients and correlated with higher hemoglobin level and higher leukocyte count. Int J Gerontol. 2013;7(1):40–4.CrossRefGoogle Scholar
Li S, Kralovics R, De Libero G, Theocharides A, Gisslinger H, Skoda RC. Clonal heterogeneity in polycythemia vera patients with JAK2 exon12 and JAK2-V617F mutations. Blood. 2008;111(7):3863–6.CrossRefPubMedGoogle Scholar
Lim KH, Lin HC, Chen CG, Wang WT, Chang YC, Chiang YH, et al. Rapid and sensitive detection of CALR exon 9 mutations using high-resolution melting analysis. Clin Chim Acta. 2015;440:133–9.CrossRefPubMedGoogle Scholar
Chao A, Chang TC, Lapke N, Jung SM, Chi P, Chen CH, et al. Prevalence and clinical significance of BRCA1/2 germline and somatic mutations in Taiwanese patients with ovarian cancer. Oncotarget. 2016;7(51):85529–41.PubMedGoogle Scholar