JAK2V617F Allele Burden Measurement in Peripheral Blood of Iranian Patients with Myeloproliferative Neoplasms and Effect of Hydroxyurea on JAK2V617F Allele Burden


Background: Myeloproliferative neoplasms (MPNs) are clonal malignant diseases that represent a group of conditions including polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). The aim of this study was to evaluate possible correlations between JAK2V617F allele burden and clinicohematologic characteristics in Iranian patients with MPNs. We also aimed at determining the correlation between JAK2V617F allele burden and use of cytoreductive treatment (hydroxyurea).
Materials and Methods: We performed ARMS-PCR for all MPNs samples and subsequently performed real-time quantitative polymerase chain reaction (qRT-PCR) for JAK2V617F allele burden measurement using DNA from peripheral blood leukocytes.
Results: Two distinct groups of patients were examined at a single time point: group A (n=40; 20 PV, 20 ET) was examined at the time of diagnosis; group B (n=85; 40 PV, 30 ET and 15 PMF) while under treatment with hydroxyurea (HU). The median allele burden of the JAK2 V617F was 72% for PV and 49% for ET patients at the time of diagnosis (p=0.01). For patients with HU treatment, we determined the median JAK2V617F allele burden to be 43%, 40%, and 46.5 % in PV, ET and PMF patients; respectively. HU-treated PV patients had a significant lower %JAK2V617F than PV patients at the time of diagnosis (43% vs. 72%, p=0.005). In ET group, the relationship between the JAK2 V617F allele burden and leukocyte count was significant (p=0.02 and p=0.01 in untreated and treated patients, respectively).
Conclusions: Our results showed that patients with PV have a higher JAK2V617F allele burden. Moreover, our study demonstrated that the JAK2V617F allele burden correlates with clinical features in ET group. We also showed hydroxyurea can affect the JAK2V617F allele burden in PV patients.

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.

Vainchenker W, Delhommeau F, Constantinescu SN, et al. New mutations and pathogenesis of myeloproliferative neoplasms. Blood. 2011; 118(7):1723-35.

Vannucchi AM, Guglielmelli P, Tefferi A. Advances in understanding and management of myeloproliferative neoplasms. CA cancer Journal Clin. 2009; 59(3):171-91.

Rumi E, Pietra D, Ferretti V, et al. JAK2 or CALR mutation status defines subtypes of essential thrombocythemia with substantially different clinical course and outcomes. Blood. 2014; 123(10):1544-51.

Edahiro Y, Morishita S, Takahashi K, et al. JAK2V617F mutation status and allele burden in classical Ph-negative myeloproliferative neoplasms in Japan. Int J Hematol. 2014; 99(5):625-34.

Kim HR, Choi HJ, Kim YK, et al. Allelic expression imbalance of JAK2 V617F mutation in BCR-ABL negative myeloproliferative neoplasms. PloS One. 2013; 8(1): e52518.

Kuriakose E, Vandris K, Wang YL, et al. Decrease in JAK2V617F allele burden is not a prerequisite to clinical response in patients with polycythemia vera. Haematologica. 2012; 97(4):538-42.

Ha JS, Kim YK, Jung SI, et al. Correlations between Janus kinase 2 V617F allele burdens and clinicohematologic parameters in myeloproliferative neoplasms. Ann Lab Med. 2012 Nov; 32(6):385-91.

Zhou J, Ye Y, Zeng S, et al. Impact of JAK2 V617F Mutation on Hemogram Variation in Patients with Non-Reactive Elevated Platelet Counts. PloS One. 2013; 8(2):e57856.

Park SH, Chi HS, Cho YU, et al. The allele burden of JAK2 V617F can aid in differential diagnosis of Philadelphia Chromosome-Negative Myeloproliferative Neoplasm. Blood Res. 2013; 48(2):128-32.

Barbui T. First-Line Therapy and Special Issues Management in Polycythemia Vera and Essential Thrombocythemia. Myeloproliferative Neoplasms: Springer. 2011. p. 93-104.

Besses C, Álvarez‐Larrán A, Martínez‐Avilés L, et al. Modulation of JAK2 V617F allele burden dynamics by hydroxycarbamide in polycythaemia vera and essential thrombocythaemia patients. Br J Haematol. 2011; 152(4):413-9.

Ricksten A, Palmqvist L, Johansson P, et al. Rapid decline of JAK2V617F levels during hydroxyurea treatment in patients with polycythemia vera and essential thrombocythemia. Haematologica. 2008; 93(8):1260-1.

Larsen TS, Pallisgaard N, de Stricker K, et al. Limited efficacy of hydroxyurea in lowering of the JAK2 V617F allele burden. Hematology. 2009;14(1):11-5.

Spanoudakis E, Bazdiara I, Kotsianidis I, et al. Hydroxyurea (HU) is effective in reducing JAK2V617F mutated clone size in the peripheral blood of essential thrombocythemia (ET) and polycythemia vera (PV) patients. Ann Hematol. 2009; 88(7):629-32.

Antonioli E, Carobbio A, Pieri L, et al. Hydroxyurea does not appreciably reduce JAK2 V617F allele burden in patients with polycythemia vera or essential thrombocythemia. Haematologica. 2010 Aug; 95(8):1435-8.

Zalcberg IR, Ayres-Silva J, de Azevedo AM, et al. Hydroxyurea dose impacts hematologic parameters in polycythemia vera and essential thrombocythemia but does not appreciably affect JAK2-V617F allele burden. Haematologica. 2011; 96(3):e18-e20.

Michiels JJ, Juvonen E, editors. Proposal for revised diagnostic criteria of essential thrombocythemia and polycythemia vera by the Thrombocythemia Vera Study Group. Semin Thromb Hemost; 1997: 23(4):339-47.

Jones AV, Kreil S, Zoi K, et al . Widespread occurrence of the JAK2 V617F mutation in chronic myeloproliferative disorders. Blood. 2005; 106(6): 2162–8.

Fantasia F, Di Capua EN, Cenfra N, et al. A highly specific q-RT-PCR assay to address the relevance of the JAK2WT and JAK2V617F expression levels and control genes in Ph-negative myeloproliferative neoplasms. Ann Hematol. 2014; 93(4):609-16.

Vannucchi A, Pancrazzi A, Bogani C, et al. A quantitative assay for JAK2V617F mutation in myeloproliferative disorders by ARMS-PCR and capillary electrophoresis. Leukemia. 2006; 20(6):1055-60.

Sazawal S, Bajaj J, Chikkara S, et al. Prevalence of JAK2 V617F mutation in Indian patients with chronic myeloproliferative disorders. Indian J Med Res. 2010; 132:423-7.

da Silva RR, Domingues HB, Machado C, et al. JAK2 V617F mutation prevalence in myeloproliferative neoplasms in Pernambuco, Brazil. Genet Test Mol Biomarkers. 2012;16(7):802-5.

Ho C-L, Wu Y-Y, Hung H-M, et al. Rapid identification of heterozygous or homozygous JAK2 V617F mutations in myeloproliferative neoplasms using melting curve analysis. Journal of the Formosan Medical Association. 2012; 111(1):34-40.

Karkucak M, Yakut T, Ozkocaman V, et al. Evaluation of the JAK2-V617F gene mutation in Turkish patients with essential thrombocythemia and polycythemia vera. Mol Biol Rep. 2012; 39(9):8663-7.

Hamidah N, Farisah N, Azlinda A, et al. A study of JAK2 (V617F) gene mutation in patients with chronic myeloproliferative disorders. La Clinica Terapeutica. 2011; 163(2):109-13.

Mahfouz RA, Hoteit R, Salem Z, et al. JAK2 V617F gene mutation in the laboratory work-up of myeloproliferative disorders: experience of a major referral center in Lebanon. Genet Test Mol Biomarkers. 2011; 15(4):263-5.

Chao H, Shen Y, Zhang R, et al. A quantitative assay for JAK2 mutation in 135 patients with chronic myeloproliferative neoplasms. Zhonghua xue ye xue za zhi= Zhonghua xueyexue zazhi. 2009;30(5):321-5.

Baxter EJ, Scott LM, Campbell PJ, et al. Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. The Lancet. 2005; 365(9464):1054-61.

Zhao R, Xing S, Li Z, et al. Identification of an acquired JAK2 mutation in polycythemia vera. J Biol Chem. 2005; 280(24):22788-92.

James C, Ugo V, Le Couédic JP, et al. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005; 434(7037):1144-8.

Levine RL, Wadleigh M, Cools J, 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.

Antonioli E, Guglielmelli P, Poli G, et al. Influence of JAK2V617F allele burden on phenotype in essential thrombocythemia. Haematologica. 2008; 93(1):41-8.

Vannucchi A, Antonioli E, Guglielmelli P, et al. Clinical correlates of JAK2V617F presence or allele burden in myeloproliferative neoplasms: a critical reappraisal. Leukemia. 2008; 22(7):1299-307.

Vannucchi A, Antonioli E, Guglielmelli P, et al. Prospective identification of high-risk polycythemia vera patients based on JAK2V617F allele burden. Leukemia. 2007; 21(9):1952-9.

Kittur J, Knudson RA, Lasho TL, et al. Clinical correlates of JAK2V617F allele burden in essential thrombocythemia. Cancer. 2007; 109(11):2279-84.

Alshemmari SH, Rajaan R, Ameen R, et al. JAK2V617F allele burden in patients with myeloproliferative neoplasms. Ann Hematol. 2014; 93(5):791-6.

Silver RT, Vandris K, Wang YL, et al. JAK2 V617F allele burden in polycythemia vera correlates with grade of myelofibrosis, but is not substantially affected by therapy. Leuk Res. 2011; 35(2):177-82.

Tefferi A, Lasho TL, Schwager SM, et al. The clinical phenotype of wild‐type, heterozygous, and homozygous JAK2V617F in polycythemia vera. Cancer. 2006; 106(3): 631-5.

Vannucchi AM, Antonioli E, Guglielmelli P, et al. Clinical profile of homozygous JAK2 617V> F mutation in patients with polycythemia vera or essential thrombocythemia. Blood. 2007; 110(3):840-6.

Passamonti F, Rumi E, Pietra D, et al. A prospective study of 338 patients with polycythemia vera: the impact of JAK2 (V617F) allele burden and leukocytosis on fibrotic or leukemic disease transformation and vascular complications. Leukemia. 2010; 24(9):1574-9.

Tefferi A, Strand J, Lasho T, et al. Bone marrow JAK2V617F allele burden and clinical correlates in polycythemia vera. Leukemia. 2007; 21(9):2074-5.

Girodon F, Schaeffer C, Cleyrat C, et al. Frequent reduction or absence of detection of the JAK2-mutated clone in JAK2V617F-positive patients within the first years of hydroxyurea therapy. Haematologica. 2008; 93(11): 1723-7.

Hussein K, Bock O, Theophile K, et al. JAK2( V617F) allele burden discriminates essential thrombocythemia from a subset of prefibrotic-stage primary myelofibrosis. Exp Hematol. 2009; 37(10):1186-93. e7.

Stein BL, Williams DM, Wang NY, et al. Sex differences in the JAK2V617F allele burden in chronic myeloproliferative disorders. Haematologica. 2010; 95(7):1090-7.

Larsen TS, Pallisgaard N, Møller MB, et al. The JAK2 V617F allele burden in essential thrombocythemia, polycythemia vera and primary myelofibrosis–impact on disease phenotype. Eur J Haematol. 2007;79(6):508-15.

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Hydroxyurea JAK2V617F Myeloproliferative neoplasms

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Ferdowsi S, Ghaffari SH, Amirizadeh N, Azarkeivan A, Atarodi K, Faranoush M, Toogeh G, Shirkoohi R, Vaezi M, Maghsoodlu M, Alimoghaddam K, Ghavamzadeh A, Teimori Naghadeh H. JAK2V617F Allele Burden Measurement in Peripheral Blood of Iranian Patients with Myeloproliferative Neoplasms and Effect of Hydroxyurea on JAK2V617F Allele Burden. Int J Hematol Oncol Stem Cell Res. 2016;10(2):70-78.