Main Article Content
Aims: Development of a JAK2 Allelic burden estimation kit using Taqman probes and determine trend in shifting of JAK2 V617F allelic burden in and its use in analysis of patients with essential thrombocythemia (ET) and polycythemia vera (PV) to predict thrombotic complications.
Methodology: Through a retrospect study, a total of 412 ET and PV patient, divided into 3 groups (0-2, 2-5 and 5-10 years) based on time of detection of the disease, were tested in retrospect for presence of the JAK2 V617F mutant allele burden using an in-house developed Taqman probe-based kit, trend in shifting of the mutant allele burden was studied and segregated into ET (n=167) and PV (n=126) group based on their disease profile. It was then categorized into 3 time periods (0-2, 2-5 and 5-10 years) based on time of detection of the disease.
Results: Around 293 (71%) were positive for JAK2 V617F while 59 (14.2%) and 8 (1.9%) positive for CALR exon 9 frame shift & MPL mutations (W515L/W515K) respectively. The 1-25% allelic burden group size gradually fell in ET population over time and this trend continued in the PV population also. In the former the fall was 7% & 11% for 2-5 & 5-10 years category while in the later, it was 1% and 15% respectively for the same time period-category.
Conclusions: There is a distinct molecular continuum in the JAK2 V617F allelic burden in the ET & PV patients which followed a predictable trend and was associated with increasingly complicated vascular events.
de Freitas RM, da Costa Maranduba CM. Myeloproliferative neoplasms and the JAK/STAT signaling pathway: An overview. Revista brasileira de hematologiae hemoterapia. 2015;37(5): 348–353.
Dameshek W. Editorial: Some speculations on the myeloproliferative syndromes. Blood. 1951;6(4):372–5.
Spivak JL, Barosi G, Tognoni G, Barbui T, Finazzi G, Marchioli R, et al. Chronic myeloproliferative disorders. Hematology-American Society of Hematology Education Program. 2003;(1):200–24.
Finazzi G, Barbui T. How I treat patients with polycythemia vera. Blood. 2007; 109(12):5104–11.
Haferlach T, Bacher U, Kern W, Schnittger S, Haferlach C. The diagnosis of BCR/ABL-negative chronic myeloprolifera-tive diseases (CMPD): A comprehensive approach based on morphology, cytogenetics, and molecular markers. Annals of Hematology. 2008;87(1):1–10.5.
Singdong R, Siriboonpiputtana T, Chareonsirisuthigul T, Kongruang A, Limsuwanachot N, Sirirat T, et al. Characterization and prognosis significance of JAK2 (V617F), MPL and CALR mutations in Philadelphia-Negative Myeloproliferative Neoplasms. Asian Pacific Journal of Cancer Prevention. 2016;17(10):4647–53.
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;365:1054-61.
James C, Ugo V, Le Couédic JP, Staerk J, Delhommeau F, Lacout C, et al. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005;434: 1144-8.
Kralovics R, Passamonti F, Buser AS, Teo SS, Tiedt R, Passweg JR, et al. A gain-of-function mutation of JAK2 in myeloproliferative disorders. The New England Journal of Medicine. 2005;352: 1779-90.
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: 387-97.
Tefferi A, Thiele J, Orazi A, Kvasnicka HM, Barbui T, Hanson CA, et al. Proposals and rationale for revision of the World Health Organization diagnostic criteria for polycythemia vera, essential thrombocythemia, and primary myelofibrosis: Recommendations from an ad hoc international expert panel. Blood. 2007;110:1092-7.
Chen Q, Lu P, Jones AV, Cross NC, Silver, RT, Wang YL. Amplification refractory mutation system, a highly sensitive and simple polymerase chain reaction assay, for the detection of JAK2 V617F mutation in chronic myeloproliferative disorders. The Journal of Molecular Diagnostics. 2007; 9(2):272–276.
Tefferi A, Lasho TL, Schwager SM, Steensma DP, Mesa RA, Li CY .The JAK2(V617F) tyrosine kinase mutation in myelofibrosis with myeloid metaplasia: Lineage specificity and clinical correlates. British Journal of Haematology. 2005;131: 320–328.
Campbell PJ, Scott LM, Baxter EJ, Bench AJ, Green AR, Erber WN. Methods for the detection of the JAK2 V617F mutation in human myeloproliferative disorders. Methods in Molecular Medicine. 2006;125: 253-64.
Passamonti F, Rumi E, Pietra D, Della Porta MG, Boveri E, Pascutto C, et al. Relation between JAK2 (V617F) mutation status, granulocyte activation, and constitutive mobilization of CD34+ cells into peripheral blood in myeloproliferative disorders. Blood. 2006;107:3676-82.
Tefferi A, Strand JJ, Lasho TL, Knudson RA, Finke CM, Gangat N, et al. Bone marrow JAK2 V617F allele burden and clinical correlates in polycythemia vera. Leukemia. 2007;21:2074-5.
Kittur J, Knudson RA, Lasho TL, Finke CM, Gangat N, Wolanskyj AP, et al. Clinical correlates of JAK2 V617F allele burden in essential thrombocythemia. Cancer. 2007;109:2279-84.
Hussein K, Bock O, Theophile K, von Neuhoff N, Buhr T, Schlué J, et al. JAK2(V617F) allele burden discriminates essential thrombocythemia from a subset of prefibrotic-stage primary myelofibrosis. Experimental Hematology. 2009;37:1186-93.
Hammond E, Shaw K, Carnley B, P'ng S, James I, Herrmann R. Quantitative determination of JAK2 V617F by TaqMan: An absolute measure of averaged copies per cell that may be associated with the different types of myeloproliferative disorders. The Journal of Molecular Diagnostics. 2007;9(2):242-8.
Kiladjian J-J, Gardin C, Renoux M, Bruno F, Bernard J-F. Long-term outcomes of polycythemia vera patients treated with pipobroman as initial therapy. Journal of Hematology 2003;4:198–207.
Mara Jorgelina Ojeda, Irma Margarita Bragós, Karina Lucrecia Calvo, Gladis Marcela Williams, María Magdalena Carbonell & Arianna Flavia Pratti. CALR, JAK2 and MPL mutation status in Argentinean patients with BCR-ABL1- negative myeloproliferative neoplasms, Hematology. 2018;23(4):208-211
Klampfl T, Gisslinger H, Harutyunyan AS, Nivarthi H, Rumi E, Milosevic JD, et al. Somatic mutations of calreticulin in myeloproliferative neoplasms. The New England Journal of Medicine. 2013;369: 2379–2390.
Berglund S, Zettervall O. Incidence of polycythemia vera in a defined population. European Journal of Haematology. 1992; 48:20-26.
Taylor KM, Shetta M, Talpaz M, Kantarjian HM, Hardikar S, Chinault AC, et al. Myeloproliferative disorders: usefulness of X-linked probes in diagnosis. Leukemia. 1989; 3:419-422.
Janssen JW, Anger BR, Drexler HG, Bartram CR, Heimpel H. Essential thrombocythemia in two sisters originating from different stem cell levels. Blood. 1990; 75:1633-1636.
Shih LY, Lee CT. Identification of masked polycythemia vera from patients with idiopathic marked thrombocytosis by endogenous erythroid colony assay. Blood. 1994;83:744-748.
Ellis JT, Peterson P, Geller SA, Rappaport H. Studies of the bone marrow in polycythemia vera and the evolution of myelofibrosis and second hematologic malignancies. Seminars in Hematology. 1986;23:144-155.
Barosi G, Cazzola M, Frassoni F, Orlandi E, Stefanelli M. Erythropoiesis in myelofibrosis with myeloid metaplasia: recognition of different classes of patients by erythrokinetics. British Journal of Haematology. 1981;48:263-272.
Cervantes F, Alvarez-Larran A, Talarn C, Gomez M, Montserrat E. Myelofibrosis with myeloid metaplasia following essential thrombocythaemia: actuarial probability, presenting characteristics and evolution in a series of 195 patients. British Journal of Haematology. 2002;118:786-790.
Bouroncle B, Doan CA. Myelofibrosis: Clinical, hematologic and pathologic study of 110 patients. The American Journal of the Medical Sciences. 1962;243:697- 715.
Pitcock JA, Reinhard EH, Justus BW, Mendelsohn RS. A clinical and pathological study of seventy cases of myelofibrosis. Annals of Internal Medicine. 1962;57:73-84.
Pettit JE, Lewis SM, Nicholas AW. Transitional myeloproliferative disorder. British Journal of Haematology. 1979;43: 167-184
Barosi G, Baraldi A, Cazzola M, Fortunato A, Palestra P, Polino G, et al. Polycythaemia following splenectomy in myelofibrosis with myeloid metaplasia: a reorganization of erythropoiesis. Scandinavian Journal of Haematology. 1984;32:12-18.
Jacobson RJ, Salo A, Fialkow PJ. Agnogenic myeloid metaplasia: A clonal proliferation of hematopoietic stem cells with secondary myelofibrosis. Blood. 1978; 51:189-194.
Fialkow PJ, Faguet GB, Jacobson RJ, Vaidya K, Murphy S. Evidence that essential Thrombocythemia is a clonal disorder with origin in a multipotent stem cell. Blood. 1981; 58: 916-919.
Huang LJ, Constantinescu SN, Lodish HF. The N-terminal domain of Janus kinase 2 is required for Golgi processing and cell surface expression of erythropoietin receptor. Molecular Cell. 2001;8:1327-1338.
Royer Y, Staerk J, Costuleanu M, Courtoy PJ, Constantinescu SN. Janus kinases affect thrombopoietin receptor cell surface localization and stability. The Journal of Biological Chemistry. 2005;280:27251-27261.
Moliterno AR, Williams DM, Rogers O, Spivak JL. Molecular mimicry in the chronic myeloproliferative disorders: reciprocity between quantitative JAK2 V617F and Mpl expression. Blood. 2006; 108:3913-3915.
Cassinat B, Laguillier C, Gardin C, de Beco V, Burcheri S, Fenaux P, et al. Classification of myeloproliferative disorders in the JAK2 era: Is there a role for red cell mass? Leukemia. 2008;22: 452-453.
Rozman C, Giralt M, Feliu E, Rubio D, Cortes MT. Life expectancy of patients with chronic nonleukemic myeloproliferative disorders. Cancer. 1991;67:2658-2663.
Vannucchi AM, Antonioli E, Guglielmelli P, Rambaldi A, Barosi G, Marchioli R, et al. Clinical profile of homozygous JAK2 V617F mutation in patients with polycythemia vera or essential thrombocythemia. Blood. 2007;110:840-846.
Videbaek A. Polycythemia vera: Course and prognosis. Acta Medica Scandinavica. 1950;138:179-187.
Scott LM, Tong W, Levine RL, Scott MA, Beer PA, Stratton MR, et al. JAK2 exon 12 mutations in polycythemia vera and idiopathic erythrocytosis. The New England Journal of Medicine. 2007;356: 459–468.
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 Medicine. 2006;3(7): e270.
Wang WA, Groenendyk J, Michalak M. Calreticulin signalling in health and disease. The International Journal of Biochemistry & Cell Biology. 2012;44(6): 842–846.
Nangalia J, Massie C.E, Baxter E.J, Nice F.L, Gundem G, Wedge D.C, et al. Somatic CALR mutations in myeloproliferative neoplasms with non-mutated JAK2. The New England Journal of Medicine. 2013;369:2391–2405.
Campbell PJ, Scott LM, Buck G, Wheatley K, East CL, Marsden JT, et al. Definition of subtypes of essential thrombocythaemia and relation to polycythaemia vera based on JAK2 V617F mutation status: A prospective study. Lancet 2005;366: 1945–1953.
Pardanani A, Fridley BL, Lasho TL, Gilliland DG, Tefferi A. Host genetic variation contributes to phenotypic diversity in myeloproliferative disorders. Blood 2008;111:2785–2789.
Vannucchi AM, Antonioli E, Guglielmelli P, Pardanani A, Tefferi A. Clinical correlates of JAK2 V617F presence or allele burden in myeloproliferative neoplasms: A critical reappraisal. Leukemia 2008;22:1299–1307.
Carobbio A, Finazzi G, Antonioli E, Guglielmelli P, Vannucchi A, Dellacasa C, et al., Silvia Salmoiraghi, Federica Delaini, Alessandro Rambaldi, and Tiziano Barbui. JAK2 V617F allele burden and thrombosis: a direct comparison in essential thrombocythemia and polycythemia vera. Experimental Hematology. 2009;37(9): 1016–1021.
Gale RE, Allen AJ, Nash MJ, Linch DC. Long-term serial analysis of X-chromosome inactivation patterns and JAK2 V617F mutant levels in patients with essential thrombocythemia show that minor mutant-positive clones can remain stable for many years. Blood. 2007;109: 1241–1243.
Theocharides A, Passweg JR, Medinger M, Looser R, Li S, Hao-Shen H, et al. The allele burden of JAK2 mutations remains stable over several years in patients with myeloproliferative disorders. Haematologica. 2008;93:1890–1893.