Reduced Erg Dosage Impairs Survival of Hematopoietic Stem and Progenitor Cells

Stem Cells. 2017 Jul;35(7):1773-1785. doi: 10.1002/stem.2627. Epub 2017 Apr 24.

Abstract

ERG, an ETS family transcription factor frequently overexpressed in human leukemia, has been implicated as a key regulator of hematopoietic stem cells. However, how ERG controls normal hematopoiesis, particularly at the stem and progenitor cell level, and how it contributes to leukemogenesis remain incompletely understood. Using homologous recombination, we generated an Erg knockdown allele (Ergkd ) in which Erg expression can be conditionally restored by Cre recombinase. Ergkd/kd animals die at E10.5-E11.5 due to defects in endothelial and hematopoietic cells, but can be completely rescued by Tie2-Cre-mediated restoration of Erg in these cells. In Ergkd/+ mice, ∼40% reduction in Erg dosage perturbs both fetal liver and bone marrow hematopoiesis by reducing the numbers of Lin- Sca-1+ c-Kit+ (LSK) hematopoietic stem and progenitor cells (HSPCs) and megakaryocytic progenitors. By genetic mosaic analysis, we find that Erg-restored HSPCs outcompete Ergkd/+ HSPCs for contribution to adult hematopoiesis in vivo. This defect is in part due to increased apoptosis of HSPCs with reduced Erg dosage, a phenotype that becomes more drastic during 5-FU-induced stress hematopoiesis. Expression analysis reveals that reduced Erg expression leads to changes in expression of a subset of ERG target genes involved in regulating survival of HSPCs, including increased expression of a pro-apoptotic regulator Bcl2l11 (Bim) and reduced expression of Jun. Collectively, our data demonstrate that ERG controls survival of HSPCs, a property that may be used by leukemic cells. Stem Cells 2017;35:1773-1785.

Keywords: Animal models; Apoptosis; Hematopoietic progenitors; Hematopoietic stem cells; Leukemia; Transcription factors.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antimetabolites / pharmacology
  • Apoptosis / drug effects
  • Apoptosis / genetics*
  • Bcl-2-Like Protein 11 / genetics
  • Bcl-2-Like Protein 11 / metabolism
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / metabolism
  • Female
  • Fluorouracil / pharmacology
  • Gene Dosage*
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Genetic Complementation Test
  • Hematopoiesis / drug effects
  • Hematopoiesis / genetics*
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / metabolism*
  • Integrases / genetics
  • Integrases / metabolism
  • Male
  • Megakaryocytes / cytology
  • Megakaryocytes / drug effects
  • Megakaryocytes / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Oncogene Proteins / deficiency
  • Oncogene Proteins / genetics*
  • Proto-Oncogene Proteins c-jun / genetics
  • Proto-Oncogene Proteins c-jun / metabolism
  • Signal Transduction
  • Transcriptional Regulator ERG / deficiency
  • Transcriptional Regulator ERG / genetics*

Substances

  • Antimetabolites
  • Bcl-2-Like Protein 11
  • Bcl2l11 protein, mouse
  • ERG protein, mouse
  • Oncogene Proteins
  • Proto-Oncogene Proteins c-jun
  • Transcriptional Regulator ERG
  • Cre recombinase
  • Integrases
  • Fluorouracil