Annals of Hematology

, Volume 96, Issue 5, pp 725–732 | Cite as

Clinicopathological differences exist between CALR- and JAK2-mutated myeloproliferative neoplasms despite a similar molecular landscape: data from targeted next-generation sequencing in the diagnostic laboratory

  • Rishu Agarwal
  • Piers Blombery
  • Michelle McBean
  • Kate Jones
  • Andrew Fellowes
  • Ken Doig
  • Cecily Forsyth
  • David A. Westerman
Original Article
  • 499 Downloads

Abstract

Mutations in CALR have recently been detected in JAK2-negative myeloproliferative neoplasms (MPNs) and are key pathological drivers in these diseases. CALR-mutated MPNs are shown to have numerous clinicopathological differences to JAK2-mutated MPNs. The basis of these differences is poorly understood. It is unknown whether these differences result directly from any differences in intracellular signalling abnormalities induced by JAK2/CALR mutations or whether they relate to other phenomena such as a differing spectrum of genetic lesions between the two groups. We aimed to review the clinicopathological and molecular features of CALR- and JAK2-mutated MPNs from samples referred for diagnostic testing using a custom-designed targeted next-generation sequencing (NGS) panel. Eighty-nine CALR-mutated cases were compared with 70 JAK2-mutated cases. CALR-mutated MPNs showed higher platelet counts and a female predominance as compared to JAK2-mutated MPNs in our cohort. We have also observed differences between CALR mutation subtypes in terms of disease phenotype, mutational frequency and allelic burden. Type 1 CALR mutations were found to be more common in myelofibrosis, associated with a higher frequency and number of additional mutations and a higher mutant allelic burden as compared to type 2 CALR mutations. Despite these biological differences, our molecular characterisation suggests that CALR- and JAK2-mutated MPNs are broadly similar in terms of the quantity, frequency and spectrum of co-occurring mutations and therefore observed biological differences are likely to not be heavily influenced by the nature and quantity of co-mutated genes.

Keywords

Myeloproliferative neoplasms Primary myelofibrosis Essential thrombocythaemia Calreticulin Targeted amplicon sequencing 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

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References

  1. 1.
    Swerdlow SH, Campo E, Haris NL, et al.,eds (2008) WHO classification of tumours of haemopoeitic and lymphoid tissues. 4th edn Lyon, France: International Agency for Research on Cancer PressGoogle Scholar
  2. 2.
    James C, Ugo V, Le Couedic JP, Staerk J, Delhommeau F, Lacout C, Garcon L, Raslova H, Berger R, Bennaceur-Griscelli A, Villeval JL, Constantinescu SN, Casadevall N, Vainchenker W (2005) A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature 434(7037):1144–1148. doi: 10.1038/nature03546 CrossRefPubMedGoogle Scholar
  3. 3.
    Kralovics R, Passamonti F, Buser AS, Teo SS, Tiedt R, Passweg JR, Tichelli A, Cazzola M, Skoda RC (2005) A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med 352(17):1779–1790. doi: 10.1056/NEJMoa051113 CrossRefPubMedGoogle Scholar
  4. 4.
    Nangalia J, Massie CE, Baxter EJ, Nice FL, Gundem G, Wedge DC, Avezov E, Li J, Kollmann K, Kent DG, Aziz A, Godfrey AL, Hinton J, Martincorena I, Van Loo P, Jones AV, Guglielmelli P, Tarpey P, Harding HP, Fitzpatrick JD, Goudie CT, Ortmann CA, Loughran SJ, Raine K, Jones DR, Butler AP, Teague JW, O'Meara S, McLaren S, Bianchi M, Silber Y, Dimitropoulou D, Bloxham D, Mudie L, Maddison M, Robinson B, Keohane C, Maclean C, Hill K, Orchard K, Tauro S, Du MQ, Greaves M, Bowen D, Huntly BJ, Harrison CN, Cross NC, Ron D, Vannucchi AM, Papaemmanuil E, Campbell PJ, Green AR (2013) Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2. N Engl J Med 369(25):2391–2405. doi: 10.1056/NEJMoa1312542 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Klampfl T, Gisslinger H, Harutyunyan AS, Nivarthi H, Rumi E, Milosevic JD, Them NC, Berg T, Gisslinger B, Pietra D, Chen D, Vladimer GI, Bagienski K, Milanesi C, Casetti IC, Sant'Antonio E, Ferretti V, Elena C, Schischlik F, Cleary C, Six M, Schalling M, Schonegger A, Bock C, Malcovati L, Pascutto C, Superti-Furga G, Cazzola M, Kralovics R (2013) Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med 369(25):2379–2390. doi: 10.1056/NEJMoa1311347 CrossRefPubMedGoogle Scholar
  6. 6.
    Pietra D, Rumi E, Ferretti VV, Di Buduo CA, Milanesi C, Cavalloni C, Sant'Antonio E, Abbonante V, Moccia F, Casetti IC, Bellini M, Renna MC, Roncoroni E, Fugazza E, Astori C, Boveri E, Rosti V, Barosi G, Balduini A, Cazzola M (2016) Differential clinical effects of different mutation subtypes in CALR-mutant myeloproliferative neoplasms. Leukemia 30(2):431–438. doi: 10.1038/leu.2015.277 CrossRefPubMedGoogle Scholar
  7. 7.
    Cazzola M, Kralovics R (2014) From Janus kinase 2 to calreticulin: the clinically relevant genomic landscape of myeloproliferative neoplasms. Blood 123(24):3714–3719. doi: 10.1182/blood-2014-03-530865 CrossRefPubMedGoogle Scholar
  8. 8.
    Rumi E, Pietra D, Ferretti V, Klampfl T, Harutyunyan AS, Milosevic JD, Them NC, Berg T, Elena C, Casetti IC, Milanesi C, Sant'antonio E, Bellini M, Fugazza E, Renna MC, Boveri E, Astori C, Pascutto C, Kralovics R, Cazzola M, Associazione Italiana per la Ricerca sul Cancro Gruppo Italiano Malattie Mieloproliferative I (2014) JAK2 or CALR mutation status defines subtypes of essential thrombocythemia with substantially different clinical course and outcomes. Blood 123(10):1544–1551. doi: 10.1182/blood-2013-11-539098 CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Chen CC, Gau JP, Chou HJ, You JY, Huang CE, Chen YY, Lung J, Chou YS, Leu YW, Lu CH, Lee KD, Tsai YH (2014) Frequencies, clinical characteristics, and outcome of somatic CALR mutations in JAK2-unmutated essential thrombocythemia. Ann Hematol 93(12):2029–2036. doi: 10.1007/s00277-014-2151-8 CrossRefPubMedGoogle Scholar
  10. 10.
    Rotunno G, Mannarelli C, Guglielmelli P, Pacilli A, Pancrazzi A, Pieri L, Fanelli T, Bosi A, Vannucchi AM, Associazione Italiana per la Ricerca sul Cancro Gruppo Italiano Malattie Mieloproliferative I (2014) Impact of calreticulin mutations on clinical and hematological phenotype and outcome in essential thrombocythemia. Blood 123(10):1552–1555. doi: 10.1182/blood-2013-11-538983 CrossRefPubMedGoogle Scholar
  11. 11.
    Tefferi A, Lasho TL, Finke CM, Knudson RA, Ketterling R, Hanson CH, Maffioli M, Caramazza D, Passamonti F, Pardanani A (2014) CALR vs JAK2 vs MPL-mutated or triple-negative myelofibrosis: clinical, cytogenetic and molecular comparisons. Leukemia 28(7):1472–1477. doi: 10.1038/leu.2014.3 CrossRefPubMedGoogle Scholar
  12. 12.
    Rumi E, Pietra D, Pascutto C, Guglielmelli P, Martinez-Trillos A, Casetti I, Colomer D, Pieri L, Pratcorona M, Rotunno G, Sant'Antonio E, Bellini M, Cavalloni C, Mannarelli C, Milanesi C, Boveri E, Ferretti V, Astori C, Rosti V, Cervantes F, Barosi G, Vannucchi AM, Cazzola M, Associazione Italiana per la Ricerca sul Cancro Gruppo Italiano Malattie Mieloproliferative I (2014) Clinical effect of driver mutations of JAK2, CALR, or MPL in primary myelofibrosis. Blood 124(7):1062–1069. doi: 10.1182/blood-2014-05-578435 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Vainchenker W, Delhommeau F, Constantinescu SN, Bernard OA (2011) New mutations and pathogenesis of myeloproliferative neoplasms. Blood 118(7):1723–1735. doi: 10.1182/blood-2011-02-292102 CrossRefPubMedGoogle Scholar
  14. 14.
    Lundberg P, Karow A, Nienhold R, Looser R, Hao-Shen H, Nissen I, Girsberger S, Lehmann T, Passweg J, Stern M, Beisel C, Kralovics R, Skoda RC (2014) Clonal evolution and clinical correlates of somatic mutations in myeloproliferative neoplasms. Blood 123(14):2220–2228. doi: 10.1182/blood-2013-11-537167 CrossRefPubMedGoogle Scholar
  15. 15.
    Tenedini E, Bernardis I, Artusi V, Artuso L, Roncaglia E, Guglielmelli P, Pieri L, Bogani C, Biamonte F, Rotunno G, Mannarelli C, Bianchi E, Pancrazzi A, Fanelli T, Malagoli Tagliazucchi G, Ferrari S, Manfredini R, Vannucchi AM, Tagliafico E, investigators A (2014) Targeted cancer exome sequencing reveals recurrent mutations in myeloproliferative neoplasms. Leukemia 28(5):1052–1059. doi: 10.1038/leu.2013.302 CrossRefPubMedGoogle Scholar
  16. 16.
    Tefferi A, Guglielmelli P, Lasho TL, Rotunno G, Finke C, Mannarelli C, Belachew AA, Pancrazzi A, Wassie EA, Ketterling RP, Hanson CA, Pardanani A, Vannucchi AM (2014) CALR and ASXL1 mutations-based molecular prognostication in primary myelofibrosis: an international study of 570 patients. Leukemia 28(7):1494–1500. doi: 10.1038/leu.2014.57 CrossRefPubMedGoogle Scholar
  17. 17.
    Vannucchi AM, Lasho TL, Guglielmelli P, Biamonte F, Pardanani A, Pereira A, Finke C, Score J, Gangat N, Mannarelli C, Ketterling RP, Rotunno G, Knudson RA, Susini MC, Laborde RR, Spolverini A, Pancrazzi A, Pieri L, Manfredini R, Tagliafico E, Zini R, Jones A, Zoi K, Reiter A, Duncombe A, Pietra D, Rumi E, Cervantes F, Barosi G, Cazzola M, Cross NC, Tefferi A (2013) Mutations and prognosis in primary myelofibrosis. Leukemia 27(9):1861–1869. doi: 10.1038/leu.2013.119 CrossRefPubMedGoogle Scholar
  18. 18.
    Tefferi A, Lasho TA, Guglielmelli P, Finke CM, Rotunno G, Elala Y, Pacilli A, Hanson CA, Pancrazzi A, Ketterling RP, Mannarelli C, Barraco D, Fanelli T, Pardanani A, Gangat N, Vannucchi AM (2016) Targeted deep sequencing in polycythemia vera and essential thrombocythemia. Blood advances 1:1CrossRefGoogle Scholar
  19. 19.
    Guglielmelli P, Lasho TL, Rotunno G, Score J, Mannarelli C, Pancrazzi A, Biamonte F, Pardanani A, Zoi K, Reiter A, Duncombe A, Fanelli T, Pietra D, Rumi E, Finke C, Gangat N, Ketterling RP, Knudson RA, Hanson CA, Bosi A, Pereira A, Manfredini R, Cervantes F, Barosi G, Cazzola M, Cross NC, Vannucchi AM, Tefferi A (2014) The number of prognostically detrimental mutations and prognosis in primary myelofibrosis: an international study of 797 patients. Leukemia 28(9):1804–1810. doi: 10.1038/leu.2014.76 CrossRefPubMedGoogle Scholar
  20. 20.
    Araki M, Yang Y, Masubuchi N, Hironaka Y, Takei H, Morishita S, Mizukami Y, Kan S, Shirane S, Edahiro Y, Sunami Y, Ohsaka A, Komatsu N (2016) Activation of the thrombopoietin receptor by mutant calreticulin in CALR-mutant myeloproliferative neoplasms. Blood 127(10):1307–1316. doi: 10.1182/blood-2015-09-671172 CrossRefPubMedGoogle Scholar
  21. 21.
    Vannucchi AM, Pieri L, Guglielmelli P (2011) JAK2 allele burden in the myeloproliferative neoplasms: effects on phenotype, prognosis and change with treatment. Ther Adv Hematol 2(1):21–32. doi: 10.1177/2040620710394474 CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Tefferi A, Wassie EA, Guglielmelli P, Gangat N, Belachew AA, Lasho TL, Finke C, Ketterling RP, Hanson CA, Pardanani A, Wolanskyj AP, Maffioli M, Casalone R, Pacilli A, Vannucchi AM, Passamonti F (2014) Type 1 versus type 2 calreticulin mutations in essential thrombocythemia: a collaborative study of 1027 patients. Am J Hematol 89(8):E121–E124. doi: 10.1002/ajh.23743 CrossRefPubMedGoogle Scholar
  23. 23.
    Tefferi A, Lasho TL, Tischer A, Wassie EA, Finke CM, Belachew AA, Ketterling RP, Hanson CA, Pardanani AD (2014) The prognostic advantage of calreticulin mutations in myelofibrosis might be confined to type 1 or type 1-like CALR variants. Blood 124(15):2465–2466. doi: 10.1182/blood-2014-07-588426 CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Guglielmelli P, Rotunno G, Fanelli T, Pacilli A, Brogi G, Calabresi L, Pancrazzi A, Vannucchi AM (2015) Validation of the differential prognostic impact of type 1/type 1-like versus type 2/type 2-like CALR mutations in myelofibrosis. Blood Cancer J 5:e360. doi: 10.1038/bcj.2015.90 CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Cabagnols X, Defour JP, Ugo V, Ianotto JC, Mossuz P, Mondet J, Girodon F, Alexandre JH, Mansier O, Viallard JF, Lippert E, Murati A, Mozziconacci MJ, Saussoy P, Vekemans MC, Knoops L, Pasquier F, Ribrag V, Solary E, Plo I, Constantinescu SN, Casadevall N, Vainchenker W, Marzac C, Bluteau O (2015) Differential association of calreticulin type 1 and type 2 mutations with myelofibrosis and essential thrombocytemia: relevance for disease evolution. Leukemia 29(1):249–252. doi: 10.1038/leu.2014.270 CrossRefPubMedGoogle Scholar
  26. 26.
    Visconte V, Makishima H, Jankowska A, Szpurka H, Traina F, Jerez A, O'Keefe C, Rogers HJ, Sekeres MA, Maciejewski JP, Tiu RV (2012) SF3B1, a splicing factor is frequently mutated in refractory anemia with ring sideroblasts. Leukemia 26(3):542–545. doi: 10.1038/leu.2011.232 CrossRefPubMedGoogle Scholar
  27. 27.
    Jeromin S, Haferlach T, Weissmann S, Meggendorfer M, Eder C, Nadarajah N, Alpermann T, Kohlmann A, Kern W, Haferlach C, Schnittger S (2015) Refractory anemia with ring sideroblasts and marked thrombocytosis cases harbor mutations in SF3B1 or other spliceosome genes accompanied by JAK2V617F and ASXL1 mutations. Haematologica 100(4):e125–e127. doi: 10.3324/haematol.2014.119032 CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Lasho TL, Finke CM, Hanson CA, Jimma T, Knudson RA, Ketterling RP, Pardanani A, Tefferi A (2012) SF3B1 mutations in primary myelofibrosis: clinical, histopathology and genetic correlates among 155 patients. Leukemia 26(5):1135–1137. doi: 10.1038/leu.2011.320 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.Division of Cancer Medicine, Department of PathologyPeter MacCallum Cancer CentreMelbourneAustralia
  2. 2.Bioinformatics CorePeter MacCallum Cancer CentreMelbourneAustralia
  3. 3.University of MelbourneMelbourneAustralia
  4. 4.Jarrett Street Specialist CentreGosfordAustralia

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