Abstract
The zinc finger transcriptional repressor Gfi-1 has been shown to play a critical role in early granulopoiesis; however, its role in late neutrophilic development is poorly understood. We report here that forced expression of a dominant negative Gfi-1 mutant, N382S, resulted in augmented mRNA levels of eosinophil major basic protein (MBP) in myeloid cells induced with G-CSF to undergo terminal neutrophilic differentiation. MBP is a cytotoxic protein that is abundantly expressed in eosinophils, but not in neutrophils. Ectopic expression of MBP inhibited the proliferation and survival of differentiating myeloid cells in response to G-CSF. Significantly, while GFI-1 is upregulated during neutrophilic differentiation, it is rapidly downregulated upon induction of eosinophilic differentiation, which was associated with increased MBP expression. Knockdown of GFI-1 in eosinophilic cells also led to increased level of MBP mRNA. These results indicate that Gfi-1 functions to inhibit the expression of MBP and aberrant expression of MBP as a result of loss of Gfi-1 function may cause premature apoptosis of differentiating neutrophils. In contrast, the rapid downregulation of Gfi-1 during eosinophilic development may allow for abundant expression of MBP, a hallmark of eosinophilic differentiation.
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References
Moroy T. The zinc finger transcription factor Growth factor independence 1 (Gfi1). Int J Biochem Cell Biol. 2005;37(3):541–6.
Kazanjian A, Gross EA, Grimes HL. The growth factor independence-1 transcription factor: new functions and new insights. Crit Rev Oncol Hematol. 2006;59(2):85–97.
van der Meer LT, Jansen JH, van der Reijden BA. Gfi1 and Gfi1b: key regulators of hematopoiesis. Leukemia. 2010;24(11):1834–43.
Zeng H, Yucel R, Kosan C, Klein-Hitpass L, Moroy T. Transcription factor Gfi1 regulates self-renewal and engraftment of hematopoietic stem cells. EMBO J. 2004;23(20):4116–25.
Hock H, Hamblen MJ, Rooke HM, et al. Gfi-1 restricts proliferation and preserves functional integrity of haematopoietic stem cells. Natue. 2004;431(7011):1002–7.
Khandanpour C, Kosan C, Gaudreau MC, et al. Growth Factor Independence 1 (Gfi1) Protects hematopoietic stem cells against apoptosis but also prevents the development of a myeloproliferative-like disease. Stem Cells 2011;29(2):376–85.
Karsunky H, Zeng H, Schmidt T, et al. Inflammatory reactions and severe neutropenia in mice lacking the transcriptional repressor Gfi1. Nat Genet. 2002;30(3):295–300.
Hock H, Hamblen MJ, Rooke HM, et al. Intrinsic requirement for zinc finger transcription factor Gfi-1 in neutrophil differentiation. Immunity. 2003;18(1):109–20.
Zhu J, Jankovic D, Grinberg A, Guo L, Paul WE. Gfi-1 plays an important role in IL-2-mediated Th2 cell expansion. Proc Natl Acad Sci USA. 2006;103(48):18214–9.
Zarebski A, Velu CS, Baktula AM, et al. Mutations in growth factor independent-1 associated with human neutropenia block murine granulopoiesis through colony stimulating factor-1. Immunity. 2008;28(3):370–80.
de la Luz Sierra M, Sakakibara S, Gasperini P, et al. The transcription factor Gfi1 regulates G-CSF signaling and neutrophil development through the Ras activator RasGRP1. Blood. 2010;115(19):3970–9.
Person RE, Li FQ, Duan Z, et al. Mutations in proto-oncogene GFI1 cause human neutropenia and target ELA2. Nat Genet. 2003;34(3):308–12.
Zhuang D, Qiu Y, Kogan SC, Dong F. Increased C/EBPepsilon expression and premature apoptosis in myeloid cells expressing Gfi-1 N382S mutant associated with severe congenital neutropenia. J Biol Chem. 2006;281(16):10745–51.
Zweidler-Mckay PA, Grimes HL, Flubacher MM, Tsichlis PN. Gfi-1 encodes a nuclear zinc finger protein that binds DNA and functions as a transcriptional repressor. Mol Cell Biol. 1996;16(8):4024–34.
Duan Z, Zarebski A, Montoya-Durango D, Grimes HL, Horwitz M. Gfi1 coordinates epigenetic repression of p21Cip/WAF1 by recruitment of histone lysine methyltransferase G9a and histone deacetylase 1. Mol Cell Biol. 2005;25(23):10338–51.
Khanna-Gupta A, Sun H, Zibello T, 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. Blood. 2007;109(10):4181–90.
Plager DA, Weiler DA, Loegering DA, et al. Comparative structure, proximal promoter elements, and chromosome location of the human eosinophil major basic protein genes. Genomics. 2001;71(3):271–81.
Zhuang D, Qiu Y, Haque SJ, Dong F. Tyrosine 729 of the G-CSF receptor controls the duration of receptor signaling: involvement of SOCS3 and SOCS1. J Leukoc Biol. 2005;78(4):1008–15.
Anastassiadis K, Kim J, Daigle N, Sprengel R, Scholer HR, Stewart AF. A predictable ligand regulated expression strategy for stably integrated transgenes in mammalian cells in culture. Gene. 2002;298(2):159–72.
Dong F, Brynes RK, Tidow N, Welte K, Lowenberg B, Touw IP. Mutations in the gene for the granulocyte colony-stimulating-factor receptor in patients with acute myeloid leukemia preceded by severe congenital neutropenia. N Engl J Med. 1995;333(8):487–93.
Lee KH, Kinashi T, Tohyama K, et al. Different stromal cell lines support lineage-selective differentiation of the multipotential bone marrow stem cell clone LyD9. J Exp Med. 1991;173(5):1257–66.
Ward AC, Smith L, de Koning JP, van Aesch Y, Touw IP. Multiple signals mediate proliferation, differentiation, and survival from the granulocyte colony-stimulating factor receptor in myeloid 32D cells. J Biol Chem. 1999;274(21):14956–62.
Kroll SL, Barth-Baus D, Hensold JO. The carboxyl-terminal domain of the granulocyte colony-stimulating factor receptor uncouples ribosomal biogenesis from cell cycle progression in differentiating 32D myeloid cells. J Biol Chem. 2001;276(52):49410–8.
van de Geijn GJ, Gits J, Touw IP. Distinct activities of suppressor of cytokine signaling (SOCS) proteins and involvement of the SOCS box in controlling G-CSF signaling. J Leukoc Biol. 2004;76(1):237–44.
Kitabayashi I, Yokoyama A, Shimizu K, Ohki M. Interaction and functional cooperation of the leukemia-associated factors AML1 and p300 in myeloid cell differentiation. EMBO J. 1998;17(11):2994–3004.
Shimizu K, Kitabayashi I, Kamada N, et al. AML1-MTG8 leukemic protein induces the expression of granulocyte colony-stimulating factor (G-CSF) receptor through the up-regulation of CCAAT/enhancer binding protein epsilon. Blood. 2000;96(1):288–96.
Fischkoff SA. Graded increase in probability of eosinophilic differentiation of HL-60 promyelocytic leukemia cells induced by culture under alkaline conditions. Leuk Res. 1988;12(8):679–86.
Ishihara K, Hong J, Zee O, Ohuchi K. Possible mechanism of action of the histone deacetylase inhibitors for the induction of differentiation of HL-60 clone 15 cells into eosinophils. Br J Pharmacol. 2004;142(6):1020–30.
Fischkoff SA, Pollak A, Gleich GJ, Testa JR, Misawa S, Reber TJ. Eosinophilic differentiation of the human promyelocytic leukemia cell line, HL-60. J Exp Med. 1984;160(1):179–96.
Weyer K, Glerup S. Placental regulation of peptide hormone and growth factor activity by proMBP. Biol Reprod. 2011;84(6):1077–86.
De La Luz Sierra M, Gasperini P, McCormick PJ, Zhu J, Tosato G. Transcription factor Gfi-1 induced by G-CSF is a negative regulator of CXCR4 in myeloid cells. Blood. 2007;110(7):2276–85.
Plager DA, Adolphson CR, Gleich GJ. A novel human homolog of eosinophil major basic protein. Immunol Rev. 2001;179:192–202.
Popken-Harris P, Checkel J, Loegering D, et al. Regulation and processing of a precursor form of eosinophil granule major basic protein (ProMBP) in differentiating eosinophils. Blood. 1998;92(2):623–31.
Ishihara K, Satoh I, Mue S, Ohuchi K. Generation of rat eosinophils by recombinant rat interleukin-5 in vitro and in vivo. Biochim Biophys Acta. 2000;1501(1):25–32.
Acknowledgments
This work was supported in part by Grants RSG-08-307-01 (FD) from The American Cancer Society and R15HL091511 (FD) from the National Institutes of Health.
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The authors declare that they have no conflict of interest.
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Liu, Q., Dong, F. Gfi-1 inhibits the expression of eosinophil major basic protein (MBP) during G-CSF-induced neutrophilic differentiation. Int J Hematol 95, 640–647 (2012). https://doi.org/10.1007/s12185-012-1078-x
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DOI: https://doi.org/10.1007/s12185-012-1078-x