A distal single nucleotide polymorphism alters long-range regulation of the PU.1 gene in acute myeloid leukemia
Ulrich Steidl, … , Frank Griesinger, Daniel G. Tenen
Ulrich Steidl, … , Frank Griesinger, Daniel G. Tenen
Published September 4, 2007
Citation Information: J Clin Invest. 2007;117(9):2611-2620. https://doi.org/10.1172/JCI30525.
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Research Article Oncology

A distal single nucleotide polymorphism alters long-range regulation of the PU.1 gene in acute myeloid leukemia

Abstract

Targeted disruption of a highly conserved distal enhancer reduces expression of the PU.1 transcription factor by 80% and leads to acute myeloid leukemia (AML) with frequent cytogenetic aberrations in mice. Here we identify a SNP within this element in humans that is more frequent in AML with a complex karyotype, leads to decreased enhancer activity, and reduces PU.1 expression in myeloid progenitors in a development-dependent manner. This SNP inhibits binding of the chromatin-remodeling transcriptional regulator special AT-rich sequence binding protein 1 (SATB1). Overexpression of SATB1 increased PU.1 expression, and siRNA inhibition of SATB1 downregulated PU.1 expression. Targeted disruption of the distal enhancer led to a loss of regulation of PU.1 by SATB1. Interestingly, disruption of SATB1 in mice led to a selective decrease of PU.1 RNA in specific progenitor types (granulocyte-macrophage and megakaryocyte-erythrocyte progenitors) and a similar effect was observed in AML samples harboring this SNP. Thus we have identified a SNP within a distal enhancer that is associated with a subtype of leukemia and exerts a deleterious effect through remote transcriptional dysregulation in specific progenitor subtypes.

Authors

Ulrich Steidl, Christian Steidl, Alexander Ebralidze, Björn Chapuy, Hye-Jung Han, Britta Will, Frank Rosenbauer, Annegret Becker, Katharina Wagner, Steffen Koschmieder, Susumu Kobayashi, Daniel B. Costa, Thomas Schulz, Karen B. O’Brien, Roel G.W. Verhaak, Ruud Delwel, Detlef Haase, Lorenz Trümper, Jürgen Krauter, Terumi Kohwi-Shigematsu, Frank Griesinger, Daniel G. Tenen

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Figure 5

SATB1- and SNP-dependent downregulation of PU.1 in distinct myeloid progenitor subsets in vivo.

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SATB1- and SNP-dependent downregulation of PU.1 in distinct myeloid prog...
(A) wbc from fetal livers of SATB1-knockout mice were harvested and KLS cells (Lin–c-kit+Sca1+), CMPs (Lin–c-kit+Sca1–CD34lowFcγII/IIIRlow), GMPs (Lin–c-kit+Sca1–CD34+FcγII/IIIR+), and MEPs (Lin–c-kit+Sca1–CD34–FcγII/IIIR–) were separated by multicolor FACS sorting. PU.1 expression was determined by quantitative RT-PCR. GAPDH served as a control. SD is indicated by error bars (n = 3). While there are no significant changes in total wbc, KLS cells, or CMPs, PU.1 expression is markedly decreased in GMPs and MEPs of SATB1–/– mice in comparison with wild-type littermates. (B) Total BM of WT/WT (n = 16) and SNP/SNP (n = 5) patients was examined by quantitative RT-PCR. GAPDH expression served as a control. Averages and standard deviations (error bars) are shown. (C) Lin–CD34+CD38–Thy1low HSCs of WT/WT (n = 8) and SNP/SNP (n = 4) patients were separated by multicolor FACS and PU. 1 expression was determined by quantitative RT-PCR. (D) Lin–CD34+CD38+CD123+CD45RA+ GMPs of WT/WT (n = 7) and SNP/SNP patients (n = 3) were FACS sorted and PU.1 expression was measured by quantitative RT-PCR. Expression of GAPDH was used as a control. (E) Lin–CD34+CD38+CD123–CD45RA– MEPs of WT/WT (n = 7) and SNP/SNP patients (n = 3) were FACS sorted and PU.1 expression was measured by quantitative RT-PCR. Error bars represent standard deviation. Statistical significance is indicated by asterisks. P < 0.01.