Activating c-KIT mutations confer oncogenic cooperativity and rescue RUNX1/ETO-induced DNA damage and apoptosis in human primary CD34+ hematopoietic progenitors

Leukemia. 2015 Feb;29(2):279-89. doi: 10.1038/leu.2014.179. Epub 2014 Jun 4.

Abstract

The RUNX1/ETO (RE) fusion protein, which originates from the t(8;21) chromosomal rearrangement, is one of the most frequent translocation products found in de novo acute myeloid leukemia (AML). In RE leukemias, activated forms of the c-KIT tyrosine kinase receptor are frequently found, thereby suggesting oncogenic cooperativity between these oncoproteins in the development and maintenance of t(8;21) malignancies. In this report, we show that activated c-KIT cooperates with a C-terminal truncated variant of RE, REtr, to expand human CD34+ hematopoietic progenitors ex vivo. CD34+ cells expressing both oncogenes resemble the AML-M2 myeloblastic cell phenotype, in contrast to REtr-expressing cells which largely undergo granulocytic differentiation. Oncogenic c-KIT amplifies REtr-depended clonogenic growth and protects cells from exhaustion. Activated c-KIT reverts REtr-induced DNA damage and apoptosis. In the presence of activated c-KIT, REtr-downregulated DNA-repair genes are re-expressed leading to an enhancement of DNA-repair efficiency via homologous recombination. Together, our results provide new mechanistic insight into REtr and c-KIT oncogenic cooperativity and suggest that augmented DNA repair accounts for the increased chemoresistance observed in t(8;21)-positive AML patients with activated c-KIT mutations. This cell-protective mechanism might represent a new therapeutic target, as REtr cells with activated c-KIT are highly sensitive to pharmacological inhibitors of DNA repair.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / metabolism
  • Apoptosis*
  • Benzamides / administration & dosage
  • Cell Cycle
  • Cell Separation
  • Chromosomes, Human, Pair 21
  • Chromosomes, Human, Pair 8
  • Cloning, Molecular
  • Core Binding Factor Alpha 2 Subunit / genetics
  • Core Binding Factor Alpha 2 Subunit / metabolism*
  • DNA Damage*
  • DNA Repair
  • Down-Regulation
  • Enzyme Inhibitors / chemistry
  • Flow Cytometry
  • HEK293 Cells
  • Hematopoietic Stem Cells / cytology*
  • Humans
  • Imatinib Mesylate
  • Mutation
  • Oncogene Proteins, Fusion / genetics
  • Oncogene Proteins, Fusion / metabolism*
  • Phenotype
  • Piperazines / administration & dosage
  • Protein Structure, Tertiary
  • Proto-Oncogene Proteins c-kit / genetics
  • Proto-Oncogene Proteins c-kit / metabolism*
  • Pyrimidines / administration & dosage
  • RUNX1 Translocation Partner 1 Protein
  • Translocation, Genetic
  • U937 Cells

Substances

  • AML1-ETO fusion protein, human
  • Antigens, CD34
  • Benzamides
  • Core Binding Factor Alpha 2 Subunit
  • Enzyme Inhibitors
  • Oncogene Proteins, Fusion
  • Piperazines
  • Pyrimidines
  • RUNX1 Translocation Partner 1 Protein
  • Imatinib Mesylate
  • Proto-Oncogene Proteins c-kit