Calreticulin mutant mice develop essential thrombocythemia that is ameliorated by the JAK inhibitor ruxolitinib

Leukemia. 2017 May;31(5):1136-1144. doi: 10.1038/leu.2016.308. Epub 2016 Nov 3.


Mutations of calreticulin (CALR) are detected in 25-30% of patients with essential thrombocythemia (ET) or primary myelofibrosis and cause frameshifts that result in proteins with a novel C-terminal. We demonstrate that CALR mutations activated signal transducer and activator of transcription 5 (STAT5) in 293T cells in the presence of thrombopoietin receptor (MPL). Human megakaryocytic CMK11-5 cells and erythroleukemic F-36P-MPL cells with knocked-in CALR mutations showed increased growth and acquisition of cytokine-independent growth, respectively, accompanied by STAT5 phosphorylation. Transgenic mice expressing a human CALR mutation with a 52 bp deletion (CALRdel52-transgenic mice (TG)) developed ET, with an increase in platelet count, but not hemoglobin level or white blood cell count, in association with an increase in bone marrow (BM) mature megakaryocytes. CALRdel52 BM cells did not drive away wild-type (WT) BM cells in in vivo competitive serial transplantation assays, suggesting that the self-renewal capacity of CALRdel52 hematopoietic stem cells (HSCs) was comparable to that of WT HSCs. Therapy with the Janus kinase (JAK) inhibitor ruxolitinib ameliorated the thrombocytosis in TG mice and attenuated the increase in number of BM megakaryocytes and HSCs. Taken together, our study provides a model showing that the C-terminal of mutant CALR activated JAK-STAT signaling specifically downstream of MPL and may have a central role in CALR-induced myeloproliferative neoplasms.

MeSH terms

  • Animals
  • Calreticulin / genetics*
  • Cell Self Renewal
  • HEK293 Cells
  • Hematopoietic Stem Cells
  • Humans
  • Janus Kinases / antagonists & inhibitors
  • Mice
  • Mice, Transgenic
  • Myeloproliferative Disorders / chemically induced
  • Myeloproliferative Disorders / etiology
  • Nitriles
  • Protein Kinase Inhibitors / pharmacology
  • Protein Kinase Inhibitors / therapeutic use
  • Pyrazoles / pharmacology
  • Pyrazoles / therapeutic use
  • Pyrimidines
  • Receptors, Thrombopoietin
  • STAT5 Transcription Factor / metabolism
  • Thrombocythemia, Essential / drug therapy
  • Thrombocythemia, Essential / genetics


  • Calreticulin
  • Nitriles
  • Protein Kinase Inhibitors
  • Pyrazoles
  • Pyrimidines
  • Receptors, Thrombopoietin
  • STAT5 Transcription Factor
  • ruxolitinib
  • Janus Kinases