The canonical Wnt signaling pathway is mediated by interaction of β-catenin with the T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) transcription factors and subsequent transcription activation of Wnt-target genes. In the hematopoietic system, the function of the pathway has been mainly investigated by rather unspecific genetic manipulations of β-catenin that yielded contradictory results. Here, we used a mouse expressing a truncated dominant negative form of the human TCF4 transcription factor (dnTCF4) that specifically abrogates β-catenin-TCF/LEF interaction. Disruption of the β-catenin-TCF/LEF interaction resulted in the accumulation of immature cells and reduced granulocytic differentiation. Mechanistically, dnTCF4 progenitors exhibited downregulation of the Csf3r gene, reduced granulocyte colony-stimulating factor (G-CSF) receptor levels, attenuation of downstream Stat3 phosphorylation after G-CSF treatment, and impaired G-CSF-mediated differentiation. Chromatin immunoprecipitation assays confirmed direct binding of TCF/LEF factors to the promoter and putative enhancer regions of CSF3R. Inhibition of β-catenin signaling compromised activation of the emergency granulopoiesis program, which requires maintenance and expansion of myeloid progenitors. Consequently, dnTCF4 mice were more susceptible to Candida albicans infection and more sensitive to 5-fluorouracil-induced granulocytic regeneration. Importantly, genetic and chemical inhibition of β-catenin-TCF/LEF signaling in human CD34+ cells reduced granulocytic differentiation, whereas its activation enhanced myelopoiesis. Altogether, our data indicate that the β-catenin-TCF/LEF complex directly regulates G-CSF receptor levels, and consequently controls proper differentiation of myeloid progenitors into granulocytes in steady-state and emergency granulopoiesis. Our results uncover a role for the β-catenin signaling pathway in fine tuning the granulocytic production, opening venues for clinical intervention that require enhanced or reduced production of neutrophils.

Cancer Science Institute of Singapore National University of Singapore Singapore

Childhood Leukemia Investigation Prague Department of Pediatric Hematology and Oncology 2nd Faculty of Medicine Charles University Prague Prague Czech Republic

Department of Cell and Developmental Biology and

Department of Hemato oncology Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

Department of Leukocyte Cell Signaling Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

Division of BioMedical Sciences Faculty of Medicine Memorial University of Newfoundland St John's NL Canada

Faculty of Science Charles University Prague Czech Republic

Harvard Stem Cell Institute Harvard Medical School Boston MA

Institute of Hematology and Blood Transfusion Prague Czech Republic; and

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Blood. 2020 Nov 26;136(22):2487-2489. doi: 10.1182/blood.2020008667. PubMed

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