Detection of the Activating JAK2 V617F Mutation in Paraffin-embedded Trephine Bone Marrow Biopsies of Patients with Chronic Myeloproliferative Diseases

Overview

Journal J Mol Diagn

Publisher Elsevier

Specialties Molecular Biology
Pathology

Date 2006 Jul 11

PMID 16825501

Citations 5

Authors

Thomas Horn

Marcus Kremer

Tobias Dechow

Walther M Pfeifer

Birgit Geist

Michael Perker

Justus Duyster

Leticia Quintanilla-Martinez

Falko Fend

Affiliations

Soon will be listed here.

Abstract

The discovery of the activating V617F mutation in the JAK2 tyrosine kinase in a high proportion of patients with Ph- chronic myeloproliferative diseases (CMPD) represents a diagnostic breakthrough for these disorders. Trephine bone marrow biopsy is an essential part of the diagnostic workup of CMPD and represents a valuable archival source of DNA. Therefore, we studied 152 paraffin-embedded trephines with CMPD and related disorders for the presence of the V617F mutation, using both allele-specific polymerase chain reaction (PCR) and nested PCR with subsequent digestion with BsaXI. Only 6 of 152 (4%) samples were not evaluable because of poor DNA quality. The V617F mutation was detected in 27 of 28 (96%) cases of polycythemia vera, 17 of 23 (74%) cases of essential thrombocythemia, 28 of 45 (62%) cases of chronic idiopathic myelofibrosis, six of eight (75%) cases of CMPD unclassified, and two of four (50%) cases of myelodysplastic/myeloproliferative syndrome. Ph+ chronic myelogenous leukemia (four cases), reactive (secondary) erythrocytosis (14 cases), and thrombocytosis (one case) as well as normal controls (19 cases) all lacked the V617F mutation. Based on results of BsaXI digestion and sequencing, 24 of 54 (44%) evaluable V617F+ cases were considered homozygously mutated. Thus, detection of the V617F JAK2 mutation is feasible in paraffin-embedded trephine biopsies and represents a major advance in the diagnostic evaluation of CMPD.

Citing Articles

Prevalence of the frequency of JAK2 (V617F) mutation in different myeloproliferative disorders in Egyptian patients.

Ebid G, Ghareeb M, Salaheldin O, Kamel M Int J Clin Exp Pathol. 2015; 8(9):11555-9.

PMID: 26617890 PMC: 4637706.

[Bone marrow biopsy: processing and use of molecular techniques].

Quintanilla-Martinez L, Tinguely M, Bonzheim I, Fend F Pathologe. 2012; 33(6):481-9.

PMID: 23085692 DOI: 10.1007/s00292-012-1647-z.

Advantages of the quenching probe method over other PCR-based methods for detection of the JAK2 V617F mutation.

Ono A, Okuhashi Y, Takahashi Y, Itoh M, Nara N, Tohda S Oncol Lett. 2012; 4(2):205-208.

PMID: 22844354 PMC: 3402723. DOI: 10.3892/ol.2012.741.

Survival and proliferative roles of erythropoietin beyond the erythroid lineage.

Noguchi C, Wang L, Rogers H, Teng R, Jia Y Expert Rev Mol Med. 2008; 10:e36.

PMID: 19040789 PMC: 3065109. DOI: 10.1017/S1462399408000860.

JAK2 exon 12 mutations in polycythemia vera and idiopathic erythrocytosis.

Scott L, Tong W, Levine R, Scott M, Beer P, Stratton M N Engl J Med. 2007; 356(5):459-68.

PMID: 17267906 PMC: 2873834. DOI: 10.1056/NEJMoa065202.

References

Vardiman J, Lee Harris N, Brunning R . The World Health Organization (WHO) classification of the myeloid neoplasms. Blood. 2002; 100(7):2292-302. DOI: 10.1182/blood-2002-04-1199. View

Bock O, Busche G, Koop C, Schroter S, Buhr T, Kreipe H . Detection of the single hotspot mutation in the JH2 pseudokinase domain of Janus kinase 2 in bone marrow trephine biopsies derived from chronic myeloproliferative disorders. J Mol Diagn. 2006; 8(2):170-7. PMC: 1867581. DOI: 10.2353/jmoldx.2006.050064. View

Greer C, Peterson S, Kiviat N, Manos M . PCR amplification from paraffin-embedded tissues. Effects of fixative and fixation time. Am J Clin Pathol. 1991; 95(2):117-24. DOI: 10.1093/ajcp/95.2.117. View

Baxter E, Scott L, Campbell P, East C, Fourouclas N, Swanton S . Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. Lancet. 2005; 365(9464):1054-61. DOI: 10.1016/S0140-6736(05)71142-9. View

Levine R, Wadleigh M, Cools J, Ebert B, Wernig G, Huntly B . Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. Cancer Cell. 2005; 7(4):387-97. DOI: 10.1016/j.ccr.2005.03.023. View

Kralovics R, Passamonti F, Buser A, Teo S, Tiedt R, Passweg J . A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med. 2005; 352(17):1779-90. DOI: 10.1056/NEJMoa051113. View

James C, Ugo V, Le Couedic J, Staerk J, Delhommeau F, Lacout C . A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005; 434(7037):1144-8. DOI: 10.1038/nature03546. View

Kaushansky K . On the molecular origins of the chronic myeloproliferative disorders: it all makes sense. Blood. 2005; 105(11):4187-90. DOI: 10.1182/blood-2005-03-1287. View

Zhao R, Xing S, Li Z, Fu X, Li Q, Krantz S . Identification of an acquired JAK2 mutation in polycythemia vera. J Biol Chem. 2005; 280(24):22788-92. PMC: 1201515. DOI: 10.1074/jbc.C500138200. View

10.

Steensma D, Dewald G, Lasho T, Powell H, McClure R, Levine R . The JAK2 V617F activating tyrosine kinase mutation is an infrequent event in both "atypical" myeloproliferative disorders and myelodysplastic syndromes. Blood. 2005; 106(4):1207-9. PMC: 1895198. DOI: 10.1182/blood-2005-03-1183. View

11.

Thiele J, Kvasnicka H, Facchetti F, Franco V, van der Walt J, Orazi A . European consensus on grading bone marrow fibrosis and assessment of cellularity. Haematologica. 2005; 90(8):1128-32. View

12.

Jones A, Kreil S, Zoi K, Waghorn K, Curtis C, Zhang L . Widespread occurrence of the JAK2 V617F mutation in chronic myeloproliferative disorders. Blood. 2005; 106(6):2162-8. DOI: 10.1182/blood-2005-03-1320. View

13.

Jelinek J, Oki Y, Gharibyan V, Bueso-Ramos C, Prchal J, Verstovsek S . JAK2 mutation 1849G>T is rare in acute leukemias but can be found in CMML, Philadelphia chromosome-negative CML, and megakaryocytic leukemia. Blood. 2005; 106(10):3370-3. PMC: 1895065. DOI: 10.1182/blood-2005-05-1800. View

14.

Levine R, Loriaux M, Huntly B, Loh M, Beran M, Stoffregen E . The JAK2V617F activating mutation occurs in chronic myelomonocytic leukemia and acute myeloid leukemia, but not in acute lymphoblastic leukemia or chronic lymphocytic leukemia. Blood. 2005; 106(10):3377-9. PMC: 1895066. DOI: 10.1182/blood-2005-05-1898. View

15.

Fend F, Bock O, Kremer M, Specht K, Quintanilla-Martinez L . Ancillary techniques in bone marrow pathology: molecular diagnostics on bone marrow trephine biopsies. Virchows Arch. 2005; 447(6):909-19. DOI: 10.1007/s00428-005-0069-1. View

16.

Campbell P, Scott L, Buck G, Wheatley K, East C, Marsden J . Definition of subtypes of essential thrombocythaemia and relation to polycythaemia vera based on JAK2 V617F mutation status: a prospective study. Lancet. 2005; 366(9501):1945-53. DOI: 10.1016/S0140-6736(05)67785-9. View

17.

Kremer M, Spitzer M, Mandl-Weber S, Stecker K, Schmidt B, Hofler H . Discordant bone marrow involvement in diffuse large B-cell lymphoma: comparative molecular analysis reveals a heterogeneous group of disorders. Lab Invest. 2003; 83(1):107-14. DOI: 10.1097/01.lab.0000050762.61660.27. View