Calreticulin Ins5 and Del52 mutations impair unfolded protein and oxidative stress responses in K562 cells expressing CALR mutants

Sci Rep. 2019 Jul 22;9(1):10558. doi: 10.1038/s41598-019-46843-z.

Abstract

Somatic mutations of calreticulin (CALR) have been described in approximately 60-80% of JAK2 and MPL unmutated Essential Thrombocythemia and Primary Myelofibrosis patients. CALR is an endoplasmic reticulum (ER) chaperone responsible for proper protein folding and calcium retention. Recent data demonstrated that the TPO receptor (MPL) is essential for the development of CALR mutant-driven Myeloproliferative Neoplasms (MPNs). However, the precise mechanism of action of CALR mutants haven't been fully unraveled. In this study, we showed that CALR mutants impair the ability to respond to the ER stress and reduce the activation of the pro-apoptotic pathway of the unfolded protein response (UPR). Moreover, our data demonstrated that CALR mutations induce increased sensitivity to oxidative stress, leading to increase oxidative DNA damage. We finally demonstrated that the downmodulation of OXR1 in CALR-mutated cells could be one of the molecular mechanisms responsible for the increased sensitivity to oxidative stress mediated by mutant CALR. Altogether, our data identify novel mechanisms collaborating with MPL activation in CALR-mediated cellular transformation. CALR mutants negatively impact on the capability of cells to respond to oxidative stress leading to genomic instability and on the ability to react to ER stress, causing resistance to UPR-induced apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calreticulin / genetics*
  • Calreticulin / metabolism*
  • Cell Transformation, Neoplastic / genetics
  • DNA Repair / genetics
  • Down-Regulation
  • Endoplasmic Reticulum Stress / genetics
  • Gene Knockdown Techniques
  • Humans
  • INDEL Mutation*
  • K562 Cells
  • Mitochondrial Proteins / antagonists & inhibitors
  • Mitochondrial Proteins / genetics
  • Mutant Proteins / genetics
  • Mutant Proteins / metabolism
  • Oxidative Stress / genetics*
  • Phenanthrenes / pharmacology
  • Primary Myelofibrosis / genetics
  • Primary Myelofibrosis / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Superoxide Dismutase / metabolism
  • Thrombocythemia, Essential / genetics
  • Thrombocythemia, Essential / metabolism
  • Transcriptome
  • Unfolded Protein Response / genetics*

Substances

  • CALR protein, human
  • Calreticulin
  • Mitochondrial Proteins
  • Mutant Proteins
  • OXR1 protein, human
  • Phenanthrenes
  • Recombinant Proteins
  • miltirone
  • Superoxide Dismutase