We demonstrate here that lymphoid enhancer-binding factor 1 (LEF-1) mediates the proliferation, survival and differentiation of granulocyte progenitor cells. We initially documented the importance of this transcription factor in the bone marrow of individuals with severe congenital neutropenia (CN) with a 'differentiation block' at the promyelocytic stage of myelopoiesis1. LEF-1 expression was greatly reduced or even absent in CN arrested promyelocytes, resulting in defective expression of the LEF-1 target genes CCND1, MYC and BIRC5, encoding cyclin D1 (ref. 2), c-Myc3 and survivin4, respectively. In contrast, healthy individuals showed highest LEF-1 expression in promyelocytes. Reconstitution of LEF-1 in early hematopoietic progenitors of two individuals with CN corrected the defective myelopoiesis and resulted in the differentiation of these progenitors into mature granulocytes. Repression of endogenous LEF-1 by specific short hairpin RNA inhibited proliferation and induced apoptosis of CD34+ progenitors from healthy individuals and of cells from two myeloid lines (HL-60 and K562). C/EBPα, a key transcription factor in granulopoiesis5,6, was directly regulated by LEF-1. These observations indicate that LEF-1 is an instructive factor regulating neutrophilic granulopoiesis whose absence plays a critical role in the defective maturation program of myeloid progenitors in individuals with CN. NOTE: In the version of this article initially published, the DOI was incorrect. The correct DOI is 10.1038/nm1474. The error has been corrected in the HTML and PDF versions of the article.
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We thank M. Morgan for critically reading the manuscript; V. Bucan and D. Lan for sequencing, G. Asgedom and A. Müller-Brechlin for excellent technical assistance, and M. Ballmaier and C. Reimer for assistance in cell sorting. We also thank the study subjects and their families for cooperation. This work was supported by the German Network “Congenital Bone Marrow Failure Syndromes” of the Federal Ministry of Education and Research (BMBF), Deutsche Forschungsgemeinschaft (SFB 566), Madeleine-Schickedanz-Kinderkrebsstiftung, and José Carreras Leukämie-Stiftung e.V.
The authors declare no competing financial interests.
mRNA expression of secretory granule proteins; LEF-1 mRNA/protein expression as well as TCF-3,-4 mRNA expression. (PDF 220 kb)
mRNA expression of indicated genes in sorted GFP− CD34+ CN cells from the LEF-1 lv and dnLEF-1 lv experiments. (PDF 26 kb)
mRNA expression of indicated genes in sorted RFP− cells from the shRNA transduction experiments. Morphology of LEF-1 and β-catenin shRNA transduced cells. (PDF 65 kb)
Effects of LEF-1 inhibition in two myeloid cell lines HL-60 and K562. (PDF 160 kb)
mRNA expression of indicated genes in sorted RFP− cells from the shRNA transduction experiments in K562 and HL-60 cell lines. (PDF 26 kb)
mRNA expression of indicated genes in sorted GFP− cells from the experiments in LEF-1 lv transduced CD34+ cells of healthy controls. (PDF 23 kb)
CN patients' characteristics and ELA2 mutations status. (PDF 15 kb)
Oligonucleotide sequences used for NoShift assay. (PDF 12 kb)
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Skokowa, J., Cario, G., Uenalan, M. et al. LEF-1 is crucial for neutrophil granulocytopoiesis and its expression is severely reduced in congenital neutropenia. Nat Med 12, 1191–1197 (2006). https://doi.org/10.1038/nm1474
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