Mutant calreticulin knockin mice develop thrombocytosis and myelofibrosis without a stem cell self-renewal advantage

Blood. 2018 Feb 8;131(6):649-661. doi: 10.1182/blood-2017-09-806356. Epub 2017 Dec 27.

Abstract

Somatic mutations in the endoplasmic reticulum chaperone calreticulin (CALR) are detected in approximately 40% of patients with essential thrombocythemia (ET) and primary myelofibrosis (PMF). Multiple different mutations have been reported, but all result in a +1-bp frameshift and generate a novel protein C terminus. In this study, we generated a conditional mouse knockin model of the most common CALR mutation, a 52-bp deletion. The mutant novel human C-terminal sequence is integrated into the otherwise intact mouse CALR gene and results in mutant CALR expression under the control of the endogenous mouse locus. CALRdel/+ mice develop a transplantable ET-like disease with marked thrombocytosis, which is associated with increased and morphologically abnormal megakaryocytes and increased numbers of phenotypically defined hematopoietic stem cells (HSCs). Homozygous CALRdel/del mice developed extreme thrombocytosis accompanied by features of MF, including leukocytosis, reduced hematocrit, splenomegaly, and increased bone marrow reticulin. CALRdel/+ HSCs were more proliferative in vitro, but neither CALRdel/+ nor CALRdel/del displayed a competitive transplantation advantage in primary or secondary recipient mice. These results demonstrate the consequences of heterozygous and homozygous CALR mutations and provide a powerful model for dissecting the pathogenesis of CALR-mutant ET and PMF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calreticulin / genetics*
  • Cell Self Renewal / genetics*
  • Cells, Cultured
  • Hematopoietic Stem Cells / physiology*
  • Homozygote
  • Leukocytosis / genetics
  • Leukocytosis / pathology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Mutation, Missense
  • Primary Myelofibrosis / genetics*
  • Splenomegaly / genetics
  • Splenomegaly / pathology
  • Thrombocythemia, Essential / genetics
  • Thrombocythemia, Essential / pathology
  • Thrombocytosis / genetics*

Substances

  • Calreticulin