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Chronic myeloproliferative neoplasms

Knock-in of murine Calr del52 induces essential thrombocythemia with slow-rising dominance in mice and reveals key role of Calr exon 9 in cardiac development

Leukemia volume 34, pages 510–521 (2020)Cite this article

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Abstract

Frameshifting mutations (−1/+2) of the calreticulin (CALR) gene are responsible for the development of essential thrombocythemia (ET) and primary myelofibrosis (PMF). The mutant CALR proteins activate the thrombopoietin receptor (TpoR) inducing cytokine-independent megakaryocyte progenitor proliferation. Here, we generated via CRISPR/Cas9 technology two knock-in mouse models that are heterozygous for a type-I murine Calr mutation. These mice exhibit an ET phenotype with elevated circulating platelets compared with wild-type controls, consistent with our previous results showing that murine CALR mutants activate TpoR. We also show that the mutant CALR proteins can be detected in plasma. The phenotype of Calr del52 is transplantable, and the Calr mutated hematopoietic cells have a slow-rising advantage over wild-type hematopoiesis. Importantly, a homozygous state of a type-1 Calr mutation is lethal at a late embryonic development stage, showing narrowed ventricular myocardium walls, similar to the murine Calr knockout phenotype, pointing to the C terminus of CALR as crucial for heart development.

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Acknowledgements

We thank Dr Patrick Jacquemin for advice on CRISPR/Cas9 and transgenesis, Dr Pedro Gomez for support with mouse facility, Dr Nicolas Dauguet for expert flow cytometry support, Dr Jing Jing Zhu for her help with the T-cell intracellular staining assay, Lidvine Genet and Céline Mouton for expert technical and administrative support. TB was supported by a Télévie PhD Fellowship. SNC is Honorary Research Director at FRS-FNRS Belgium. Funding to SNC is acknowledged from Ludwig Institute for Cancer Research, Fondation contre le cancer, Fondation « Les avions de Sébastien », Fondation Salus Sanguinis and projects Action de Recherche Concertée (ARC) 16/21-073 and WelBio (Walloon Excellence In Life Sciences and Biotechnology F 44/8/5- MCF/UIG-10955), Brussels, Belgium.

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Authors and Affiliations

  1. Ludwig Institute for Cancer Research, Brussels, Belgium

    Thomas Balligand, Christian Pecquet, Gilles Gaudray, Didier Colau, Yacine Rahmani, Vincent Stroobant, Benoît J. Van den Eynde, Jean-Philippe Defour & Stefan N. Constantinescu

  2. de Duve Institute, Université catholique de Louvain, Brussels, Belgium

    Thomas Balligand, Christian Pecquet, Gilles Gaudray, Yacine Rahmani, Benoît J. Van den Eynde, Jean-Philippe Defour & Stefan N. Constantinescu

  3. WELBIO (Walloon Excellence in Life Sciences and Biotechnology), Brussels, Belgium

    Thomas Balligand, Christian Pecquet, Yacine Rahmani, Benoît J. Van den Eynde, Jean-Philippe Defour & Stefan N. Constantinescu

  4. de Duve Institute, LPAD Unit, Université catholique de Louvain, Brussels, Belgium

    Younes Achouri

  5. MyeloPro Research and Diagnostics GmbH, Vienna, Austria

    Eva Hug

  6. INSERM, Unité Mixte de Recherche 1170, Gustave Roussy, Villejuif, France

    Isabelle Plo & William Vainchenker

  7. Paris-Saclay, Unité Mixte de Recherche 1170, Institut Gustave Roussy, Villejuif, France

    Isabelle Plo & William Vainchenker

  8. Gustave Roussy, Mixte de Recherche 1170, Villejuif, France

    Isabelle Plo & William Vainchenker

  9. CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria

    Robert Kralovics

  10. Cliniques Universitaires de Saint-Luc, Université catholique de Louvain, Brussels, Belgium

    Jean-Philippe Defour & Stefan N. Constantinescu

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  1. Thomas Balligand
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Correspondence to Stefan N. Constantinescu.

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RK and SNC are cofounders of MyeloPro Research and Diagnostics GmbH, Vienna, Austria. The remaining authors declare that they have no conflict of interest.

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Balligand, T., Achouri, Y., Pecquet, C. et al. Knock-in of murine Calr del52 induces essential thrombocythemia with slow-rising dominance in mice and reveals key role of Calr exon 9 in cardiac development. Leukemia 34, 510–521 (2020). https://doi.org/10.1038/s41375-019-0538-1

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