Mutations in growth factor independent-1 associated with human neutropenia block murine granulopoiesis through colony stimulating factor-1.

Severe congenital neutropenia (SCN) is characterized by a deficiency of mature neutrophils, leading to recurrent bacterial and fungal infections. Although mutations in Elastase-2, neutrophil (ELA2) predominate in human SCN, mutation of Ela2 in mice does not recapitulate SCN. The growth factor independent-1 (GFI1) transcription factor regulates ELA2. Mutations in GFI1 are associated with human SCN, and genetic deletion of Gfi1 results in murine neutropenia. We examined whether human SCN-associated GFI1N382S mutant proteins are causal in SCN and found that GFI1 functions as a rate-limiting granulopoietic molecular switch. The N382S mutation inhibited GFI1 DNA binding and resulted in a dominant-negative block to murine granulopoiesis. Moreover, Gfi1N382S selectively derepressed the monopoietic cytokine CSF1 and its receptor. Gfi1N382S-expressing Csf1-/- cells formed neutrophils. These results reveal a common transcriptional program that underlies both human and murine myelopoiesis, and that is central to the pathogenesis of SCN associated with mutations in GFI1. This shared transcriptional pathway may provide new avenues for understanding SCN caused by mutations in other genes and for clinical intervention into human neutropenias.

[1]  J. Trent.  Molecular modeling of drug-DNA complexes: an update. , 2001, Methods in enzymology.

[2]  Sean R. Eddy,et al.  Multiple Alignment Using Hidden Markov Models , 1995, ISMB.

[3]  Hui Zeng,et al.  Gfi1 and Gfi1b act equivalently in haematopoiesis, but have distinct, non‐overlapping functions in inner ear development , 2006, EMBO reports.

[4]  Hui Zeng,et al.  Inflammatory reactions and severe neutropenia in mice lacking the transcriptional repressor Gfi1 , 2002, Nature Genetics.

[5]  Bengt Fadeel,et al.  HAX1 deficiency causes autosomal recessive severe congenital neutropenia (Kostmann disease) , 2007, Nature Genetics.

[6]  L. Notarangelo,et al.  G-CSF treatment of severe congenital neutropenia reverses neutropenia but does not correct the underlying functional deficiency of the neutrophil in defending against microorganisms. , 2007, Blood.

[7]  C. Pabo,et al.  Beyond the "recognition code": structures of two Cys2His2 zinc finger/TATA box complexes. , 2001, Structure.

[8]  T. Möröy,et al.  Auxiliary splice factor U2AF26 and transcription factor Gfi1 cooperate directly in regulating CD45 alternative splicing , 2006, Nature Immunology.

[9]  S. Nishikawa,et al.  The murine mutation osteopetrosis is in the coding region of the macrophage colony stimulating factor gene , 1990, Nature.

[10]  Scott Cameron,et al.  Intrinsic requirement for zinc finger transcription factor Gfi-1 in neutrophil differentiation. , 2003, Immunity.

[11]  Holger Gohlke,et al.  The Amber biomolecular simulation programs , 2005, J. Comput. Chem..

[12]  Ronald McCarthy,et al.  Mice lacking neutrophil elastase reveal impaired host defense against gram negative bacterial sepsis , 1998, Nature Medicine.

[13]  D. Zhuang,et al.  Increased CCAAT enhancer-binding protein epsilon (C/EBPepsilon) expression and premature apoptosis in myeloid cells expressing Gfi-1 N382S mutant associated with severe congenital neutropenia. , 2006, The Journal of biological chemistry.

[14]  Zhijun Duan,et al.  Targeted transcriptional repression of Gfi1 by GFI1 and GFI1B in lymphoid cells. , 2004, Nucleic acids research.

[15]  J. Jansen,et al.  Diminished proteasomal degradation results in accumulation of Gfi1 protein in monocytes. , 2007, Blood.

[16]  Fan Dong,et al.  Increased CCAAT Enhancer-binding Protein ϵ (C/EBPϵ) Expression and Premature Apoptosis in Myeloid Cells Expressing Gfi-1 N382S Mutant Associated with Severe Congenital Neutropenia* , 2006, Journal of Biological Chemistry.

[17]  P. Tsichlis,et al.  The Gfi-1 proto-oncoprotein contains a novel transcriptional repressor domain, SNAG, and inhibits G1 arrest induced by interleukin-2 withdrawal , 1996, Molecular and cellular biology.

[18]  J. Berg,et al.  A 2.2 Å resolution crystal structure of a designed zinc finger protein bound to DNA , 1996, Nature Structural Biology.

[19]  T. Möröy,et al.  High levels of the onco-protein Gfi-1 accelerate T-cell proliferation and inhibit activation induced T-cell death in Jurkat T-cells , 2002, Oncogene.

[20]  Koenraad Devriendt,et al.  Constitutively activating mutation in WASP causes X-linked severe congenital neutropenia , 2001, Nature Genetics.

[21]  P. Tsichlis,et al.  repressor . binds DNA and functions as a transcriptional Gfi-1 encodes a nuclear zinc finger protein , 1996 .

[22]  M. Simon,et al.  The Transcriptional Repressor GFI-1 Antagonizes PU.1 Activity through Protein-Protein Interaction* , 2007, Journal of Biological Chemistry.

[23]  Christine Kinnon,et al.  Two novel activating mutations in the Wiskott-Aldrich syndrome protein result in congenital neutropenia. , 2006, Blood.

[24]  D. Link,et al.  Mice expressing a neutrophil elastase mutation derived from patients with severe congenital neutropenia have normal granulopoiesis. , 2002, Blood.

[25]  Philip S Rosenberg,et al.  The incidence of leukemia and mortality from sepsis in patients with severe congenital neutropenia receiving long-term G-CSF therapy. , 2005, Blood.

[26]  P. Lásló,et al.  Multilineage Transcriptional Priming and Determination of Alternate Hematopoietic Cell Fates , 2006, Cell.

[27]  Jing Chen,et al.  GenomeTrafac: a whole genome resource for the detection of transcription factor binding site clusters associated with conventional and microRNA encoding genes conserved between mouse and human gene orthologs , 2006, Nucleic Acids Res..

[28]  George Eliopoulos,et al.  Mutations in proto-oncogene GFI1 cause human neutropenia and target ELA2 , 2003, Nature Genetics.

[29]  Fred Schaper,et al.  The zinc finger protein Gfi‐1 can enhance STAT3 signaling by interacting with the STAT3 inhibitor PIAS3 , 2000, The EMBO journal.

[30]  W. Wiktor-Jedrzejczak,et al.  Total absence of colony-stimulating factor 1 in the macrophage-deficient osteopetrotic (op/op) mouse. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[31]  R. Hawley.  High‐Titer Retroviral Vectors for Efficient Transduction of Functional Genes into Murine Hematopoietic Stem Cells a , 1994, Annals of the New York Academy of Sciences.

[32]  C. Klein,et al.  Comparative Analysis of Genetically Modified Dendritic Cells and Tumor Cells as Therapeutic Cancer Vaccines , 2000, The Journal of experimental medicine.

[33]  C. Gilks,et al.  The Gfi-1 protooncoprotein represses Bax expression and inhibits T-cell death. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Stephen J Salipante,et al.  Neutrophil elastase in cyclic and severe congenital neutropenia. , 2007, Blood.

[35]  S. Gordon,et al.  Polymorphic expression of a neutrophil differentiation antigen revealed by monoclonal antibody 7/4 , 2004, Immunogenetics.

[36]  Hui Zeng,et al.  Transcription factor Gfi1 regulates self‐renewal and engraftment of hematopoietic stem cells , 2004, The EMBO journal.

[37]  D. Hume,et al.  Mononuclear phagocyte system of the mouse defined by immunohistochemical localization of antigen F4/80. , 1983 .

[38]  Hong Sun,et al.  Growth factor independence-1 (Gfi-1) plays a role in mediating specific granule deficiency (SGD) in a patient lacking a gene-inactivating mutation in the C/EBPepsilon gene. , 2007, Blood.