Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Rare type 1-like and type 2-like calreticulin mutants induce similar myeloproliferative neoplasms as prevalent type 1 and 2 mutants in mice

Oncogene volume 38, pages 1651–1660 (2019)Cite this article

Subjects

Abstract

Frameshift mutations in the calreticulin (CALR) gene are present in 30% of essential thrombocythemia and myelofibrosis patients. The two most frequent mutations are CALR del52 (type 1, approximately 60%) and CALR ins5 (type 2, around 30%), but many other rarer mutations exist accounting each for less than 2% of all CALR mutations. Most of them are structurally classified as type 1-like and type 2-like CALR mutations according to the absence or presence of a residual wild-type calcium-binding motif and the modification of the alpha-helix structure. Yet, several key questions remain unanswered, especially the reason of such low frequencies of these other mutations. In an attempt to investigate specific pathogenic differences between type 1-like and type 2-like CALR mutations and del52 and ins5, we modeled two type 1-like (del34 and del46) and one type 2-like (del19) mutations in cell lines and in mice. All CALR mutants constitutively activate JAK2 and STAT5/3/1 in a similar way in the presence of the thrombopoietin receptor (MPL) and induced cytokine-independent cell growth but to a lesser extent with rare mutants over time. This correlates with reduced expression levels of rare CALR mutants compared to del52 and ins5. Lethally irradiated mice that were engrafted with bone marrow transduced with the different CALR mutations developed thrombocytosis, but to a much lesser extent with ins5 and the type 2-like CALR mutation. In contrast to type 2-like mice, type 1-like mice developed marked myelofibrosis and splenomegaly 10 months after engraftment. Similar to del52, type 1-like CALR mutations induced an expansion at an early stage of hematopoiesis compared to ins5 and type 2-like mutation. Thus, type 1-like and type 2-like CALR mutants structurally and functionally resemble del52 and ins5 mutants, respectively.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Barbui T, Thiele J, Gisslinger H, Finazzi G, Vannucchi AM, Tefferi A. The 2016 revision of WHO classification of myeloproliferative neoplasms: clinical and molecular advances. Blood Rev. 2016;30:453–9.

    Article  CAS  Google Scholar 

  2. Vainchenker W, Kralovics R. Genetic basis and molecular pathophysiology of classical myeloproliferative neoplasms. Blood. 2017;129:667–79.

    Article  CAS  Google Scholar 

  3. 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;369:2391–405.

    Article  CAS  Google Scholar 

  4. 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;369:2379–90.

    Article  CAS  Google Scholar 

  5. Chachoua I, Pecquet C, El-Khoury M, Nivarthi H, Albu RI, Marty C, et al. Thrombopoietin receptor activation by myeloproliferative neoplasm associated calreticulin mutants. Blood. 2016;127:1325–35.

    Article  CAS  Google Scholar 

  6. Elf S, Abdelfattah NS, Chen E, Perales-Paton J, Rosen EA, Ko A, et al. Mutant calreticulin requires both its mutant C-terminus and the thrombopoietin receptor for oncogenic transformation. Cancer Discov. 2016;6:368–81.

    Article  CAS  Google Scholar 

  7. Araki M, Yang Y, Masubuchi N, Hironaka Y, Takei H, Morishita S, et al. Activation of the thrombopoietin receptor by mutant calreticulin in CALR-mutant myeloproliferative neoplasms. Blood. 2016;127:1307–16.

    Article  CAS  Google Scholar 

  8. Marty C, Pecquet C, Nivarthi H, El-Khoury M, Chachoua I, Tulliez M, et al. Calreticulin mutants in mice induce an MPL-dependent thrombocytosis with frequent progression to myelofibrosis. Blood. 2016;127:1317–24.

    Article  CAS  Google Scholar 

  9. Elala YC, Lasho TL, Gangat N, Finke C, Barraco D, Haider M, et al. Calreticulin variant stratified driver mutational status and prognosis in essential thrombocythemia. Am J Hematol. 2016;91:503–6.

    Article  CAS  Google Scholar 

  10. Pietra D, Rumi E, Ferretti VV, Di Buduo CA, Milanesi C, Cavalloni C, et al. Differential clinical effects of different mutation subtypes in CALR-mutant myeloproliferative neoplasms. Leukemia. 2016;30:431–8.

    Article  CAS  Google Scholar 

  11. Cabagnols X, Defour JP, Ugo V, Ianotto JC, Mossuz P, Mondet J, et al. Differential association of calreticulin type 1 and type 2 mutations with myelofibrosis and essential thrombocytemia: relevance for disease evolution. Leukemia. 2015;29:249–52.

    Article  CAS  Google Scholar 

  12. Tefferi A, Wassie EA, Guglielmelli P, Gangat N, Belachew AA, Lasho TL, et al. Type 1 versus Type 2 calreticulin mutations in essential thrombocythemia: a collaborative study of 1027 patients. Am J Hematol. 2014;89:E121–4.

    Article  CAS  Google Scholar 

  13. Eder-Azanza L, Navarro D, Aranaz P, Novo FJ, Cross NC, Vizmanos JL. Bioinformatic analyses of CALR mutations in myeloproliferative neoplasms support a role in signaling. Leukemia. 2014;28:2106–9.

    Article  CAS  Google Scholar 

  14. Tefferi A, Lasho TL, Tischer A, Wassie EA, Finke CM, Belachew AA, et al. The prognostic advantage of calreticulin mutations in myelofibrosis might be confined to type 1 or type 1-like CALR variants. Blood. 2014;124:2465–6.

    Article  CAS  Google Scholar 

  15. Balligand T, Achouri Y, Pecquet C, Chachoua I, Nivarthi H, Marty C, et al. Pathologic activation of thrombopoietin receptor and JAK2-STAT5 pathway by frameshift mutants of mouse calreticulin. Leukemia. 2016;30:1775–8.

    Article  CAS  Google Scholar 

  16. Shide K, Kameda T, Yamaji T, Sekine M, Inada N, Kamiunten A, et al. Calreticulin mutant mice develop essential thrombocythemia that is ameliorated by the JAK inhibitor ruxolitinib. Leukemia. 2017;31:1136–44.

    Article  CAS  Google Scholar 

  17. Li J, Prins D, Park HJ, Grinfeld J, Gonzalez-Arias C, Loughran S, et al. Mutant calreticulin knockin mice develop thrombocytosis and myelofibrosis without a stem cell self-renewal advantage. Blood. 2018;131:649–61.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from Ligue Nationale Contre le Cancer (“Equipe labellisée 2016”, H.R.), Institut National du Cancer (PLBIO-2015, I.P.), MPN Research Foundation (I.P.), Agence Nationale de la Recherche (ANR-DFG 2014, JAKPOT, J.L.V.) and Institut National de la Santé et de la Recherche Médicale (Inserm). B.P-S. was funded by Institut de Médecine Moléculaire Personnalisée du Cancer (MMO, ANR). O.B. was supported by Agence Nationale de la Recherche. Support to S.N.C. was from Ludwig Institute for Cancer Research, Salus Sanguinis, Fondation contre le Cancer, Action de Recherche Concertée and Belgian Genetic Medical Initiative Interuniversity Attraction Pole, Belspo. We thank O. Bawa for histology, the staff of the animal facilities of Gustave Roussy directed by P. Gonin, and P. Rameau and Y. Lecluse at the Imaging and Cytometry Platform.

Author information

Author notes

  1. These authors contributed equally: Katte Rao Toppaldoddi, Maira da Costa Cacemiro

  2. These authors contributed equally: William Vainchenker, Caroline Marty

Authors and Affiliations

  1. INSERM, UMR 1170, Gustave Roussy, Villejuif, France

    Katte Rao Toppaldoddi, Maira da Costa Cacemiro, Olivier Bluteau, Barbara Panneau-Schmaltz, Amélie Pioch, Delphine Muller, Jean-Luc Villeval, Hana Raslova, Isabelle Plo, William Vainchenker & Caroline Marty

  2. Université Paris-Saclay, UMR 1170, Gustave Roussy, Villejuif, France

    Katte Rao Toppaldoddi, Maira da Costa Cacemiro, Olivier Bluteau, Barbara Panneau-Schmaltz, Amélie Pioch, Delphine Muller, Jean-Luc Villeval, Hana Raslova, Isabelle Plo, William Vainchenker & Caroline Marty

  3. Gustave Roussy, UMR 1170, Villejuif, France

    Katte Rao Toppaldoddi, Maira da Costa Cacemiro, Olivier Bluteau, Barbara Panneau-Schmaltz, Amélie Pioch, Delphine Muller, Jean-Luc Villeval, Hana Raslova, Isabelle Plo, William Vainchenker & Caroline Marty

  4. Department of Clinical Analysis, Toxicology and Food Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo-USP, Ribeirão Preto, São Paulo, Brazil

    Maira da Costa Cacemiro

  5. Ludwig Institute for Cancer Research, Brussels, Belgium

    Stefan N. Constantinescu

  6. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium

    Stefan N. Constantinescu

Authors
  1. Katte Rao Toppaldoddi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maira da Costa Cacemiro
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Olivier Bluteau
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Barbara Panneau-Schmaltz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Amélie Pioch
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Delphine Muller
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jean-Luc Villeval
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hana Raslova
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Stefan N. Constantinescu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Isabelle Plo
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. William Vainchenker
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Caroline Marty
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Caroline Marty.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Toppaldoddi, K.R., da Costa Cacemiro, M., Bluteau, O. et al. Rare type 1-like and type 2-like calreticulin mutants induce similar myeloproliferative neoplasms as prevalent type 1 and 2 mutants in mice. Oncogene 38, 1651–1660 (2019). https://doi.org/10.1038/s41388-018-0538-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-018-0538-z

This article is cited by

Search

Advanced search

Quick links