We analyzed the structure of the granulocyte colony-stimulating factor (G-CSF) receptor gene in a 6-year-old female patient with severe congenital neutropenia (SCN) who experienced severe recurrent infections since 1 month of age. There is no family history of any similar disease. When the patient was 4 months old, she began receiving treatment with recombinant human G-CSF that resulted in a small increase in the neutrophil count sufficient for the prevention and treatment of bacterial infection. An analysis of complementary DNA for the patient’s G-CSF receptor revealed a 3-base pair deletion in the juxtamembrane intracellular sequence. This deletion at the beginning of exon 16 was thought to be caused by alternative splicing; analysis of the DNA revealed a G-to-A point mutation of the final nucleotide of intron 15. To evaluate the functional activity of the G-CSF receptor with this 3-base pair deletion of the juxtamembrane region, we transfected this G-CSF receptor mutant into an interleukin 3-dependent cell line, BAF/3. BAF/3 cells expressing the mutant G-CSF receptor showed augmented proliferation activity in response to G-CSF compared with cells having the wild-type G-CSF receptor. Although the proliferation signal of G-CSF in normal hematopoiesis is transduced through the activation of MAP kinases, this G-CSF receptor mutant showed decreased activation of ERK1/2 in response to G-CSF compared with the wild type, but the transduced signal for Stat3 activation by G-CSF was of the same magnitude as that of the wild-type G-CSF receptor. This result means that the augmented proliferation activity in response to G-CSF that we observed in cells having the G-CSF receptor gene with the 3-base pair deletion is transduced through an intracellular signaling pathway other than MAP kinase. Because SCN patients with a mutation in the G-CSF receptor frequently develop leukemia, this 3-base pair deletion in the juxtamembrane sequence of the G-CSF receptor gene in this patient may be one step in the course of leukemic transformation.
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Nagata S, Tsuchiya M, Asano S, et al. Molecular cloning and expression of cDNA for human granulocyte colony-stimulating factor.Nature. 1986;319:415–418.
Souza LM, Boone TC, Gabrilove J, et al. Recombinant human granulocyte colony-stimulating factor: effects on normal and leukemic myeloid cells.Science. 1986;232:61–65.
Kostmann R. Infantile genetic agranulocytosis: agranulocytosis infantilis hereditaria.Acta Paediatr. 1956;45(suppl 105):1–78.
Kostman R. Infantile genetic agranulocytosis: a review with presentation of ten new cases.Acta Paediatr Scand. 1975;64:362–368.
Kawaguchi Y, Kobayashi M, Tanabe A, et al. Granulopoiesis in patients with congenital neutropenia.Am J Hematol. 1985;20:223–2344.
Bonilla MA, Gillio AP, Ruggeiro M, et al. Effects of recombinant human granulocyte colony-stimulating factor on neutropenia in patients with congenital agranulocytosis.N Engl J Med. 1989;320:1574–15800.
Weite K, Zeidler C, Reiter A, et al. Differential effects of granulocyte- macrophage colony-stimulating factor and granulocyte colony- stimulating factor in children with severe congenital neutropenia.Blood. 1990;75:1056–1063.
Jones EA, Bolyard AA, Dale DC. Quality of life of patients with severe chronic neutropenia receiving long-term treatment with granulocyte colony-stimulating factor.JAMA. 1993;270:1132–1133.
Glasser L, Duncan BR, Corrigan JJ Jr. Measurement of serum granulocyte colony-stimulating factor in a patient with congenital agranulocytosis (Kostmann’s syndrome).Am J Dis Child. 1991;145:925–9288.
Pietsch T, Buhrer C, Mempel K, et al. Blood mononuclear cells from patients with severe congenital neutropenia are capable of producing granulocyte colony-stimulating factor.Blood. 1991;77:1234–12377.
Kyas U, Pietsch T, Weite K. Expression of receptors for granulocyte colony-stimulating factor on neutrophils from patients with severe congenital neutropenia and cyclic neutropenia.Blood. 1992;79:1144–11477.
Tidow N, Pilz C, Kasper B, Weite K. Frequency of point mutations in the gene for the G-CSF receptor in patients with chronic neutropenia undergoing G-CSF therapy.Stem Cells. 1997;15(suppl 1):113–1199, discussion 120.
Touw IP, Dong F. Severe congenital neutropenia terminating in acute myeloid leukemia: disease progression associated with mutations in the granulocyte-colony stimulating factor receptor gene.Leuk Res. 1996;20:629–631.
Hunter MG, Avalos BR. Granulocyte colony-stimulating factor receptor mutations in severe congenital neutropenia transforming to acute myelogenous leukemia confer resistance to apoptosis and enhance cell survival.Blood. 2000;95:2132–2137.
Dale DC, Person RE, Bolyard AA, et al. Mutations in the gene encoding neutrophil elastase in congenital and cyclic neutropenia.Blood. 2000;95:2132–2137.
Horwitz M, Benson KF, Person RE, Aprikyan AG, Dale DC. Mutations in ELA2, encoding neutrophil elastase, define a 21-day biological clock in cyclic haematopoiesis.Nat Genet. 1999;23:433–436.
Fukunaga R, Ishizaka-Ikeda E, Nagata S. Growth and differentiation signals mediated by different regions in the cytoplasmic domain of granulocyte colony-stimulating factor receptor.Cell. 1993;74:1079–1087.
Mizuno Y, Hara T, Nagata M, et al. Serum granulocyte colony-stimulating factor levels in chronic neutropenia of infancy.Pediatr Hematol Oncol. 1990;7:377–381.
Freedman MH, Alter BP. Risk of myelodysplastic syndrome and acute myeloid leukemia in congenital neutropenias.Semin Hematol. 2002;39:128–133.
Fukunaga R, Ishizaka-Ikeda E, Seto Y, Nagata S. Expression cloning of a receptor for murine granulocyte colony-stimulating factor.Cell. 1990;61:341–350.
Shimoda K, Feng J, Murakami H, et al. Jakl plays an essential role for receptor phosphorylation and Stat activation in response to granulocyte colony-stimulating factor.Blood. 1997;90:597–604.
Fukunaga R, Seto Y, Mizushima S, Nagata S. Three different mRNAs encoding human granulocyte colony-stimulating factor receptor.Proc NatlAcad Sci USA. 1990;87:8702–8706.
Seto Y, Fukunaga R, Nagata S. Chromosomal gene organization od ing human granulocyte colony-stimulating factor receptor.J Immunol. 1992;148:259–266.
Shibata S, Asano Y, Yokoyama T, et al. Analysis of the granulocyte colony-stimulating factor receptor. gene structure using lethalit in myeloid leukemia and myelodysplastic syndrome.Eur J Haematol. 1998;60:197–201.
Dong F, Hoefsloot LH, Schelen AM, et al. Identification of a nonsense mutation in the granulocyte-colony-stimulating factor receptor in severe congenital neutropenia.Proc Natl Acad Sci USA. 1994;91:4480–4484.
Fukunaga R, Ishizaka-Ikeda E, Pan CX, Seto Y, Nagata S. Functional domains of the granulocyte colony-stimulating factor receptor.EMBO J. 1991;10:2855–2865.
Dong F, van BuitenenC, Pouwels K, Hoefsloot LH, Lowenberg B, Touw IP. Distinct cytoplasmic regions of the human granulocyte colony-stimulating factor receptor involved in induction of proliferation and maturation.Mol Cell Biol. 1993;13:7774–7781.
Ziegler SF, Bird TA, Morella KK, Mosley B, Gearing DP, Baumann H. Distinct regions of the human granulocyte-colony- stimulating factor receptor cytoplasmic domain are required for proliferation and gene induction.Mol Cell Biol. 1993;13:2384–23900.
Awaya N, Uchida H, Miyakawa Y, et al. Novel variant isoform of G-CSF receptor involved in induction of proliferation of FDCP-2 cells: relevance to the pathogenesis of myelodysplastic syndrome.J. Cell Physiol. 2002;191:327–335.
Dong F, Brynes RK, Tidow N, Weite 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;333:487–493.
Tidow N, Pilz C, Teichmann B, et al. Clinical relevance of point mutations in the cytoplasmic domain of the granulocyte colony- stimulating factor receptor gene in patients with severe congenital neutropenia.Blood. 1997;89:2369–2375.
Corey SJ, Burkhardt AL, Bolen JB, Geahlen RL, Tkatch LS, Tweardy DJ. Granulocyte colony-stimulating factor receptor signaling involves the formation of a three-component complex with Lyn and Syk protein-tyrosine kinases.Proc Natl Acad Sci USA. 1994;91:4683–4687.
Matsuda T, Hirano T. Association of p72 tyrosine kinase with Stat factors and its activation by interleukin-3, interleukin-6, and granulocyte colony-stimulating factor.Blood. 1994;83:3457–3461.
Weite T, Zhang SS, Wang T, et al. STAT3 deletion during hematopoiesis causes Crohn’s disease-like pathogenesis and lethality: a critical role of STAT3 in innate immunity.Proc Natl Acad Sci USA 2003;100:1879–1884.
Lee CK, Raz R, Gimeno R, et al. STAT3 is a negative regulator of granulopoiesis but is not required for G-CSF-dependent differentiation.Immunity. 2002;17:63–72.
Kamezaki K, Shimoda K, Numata A, et al. Roles of Stat3 and ERK in G-CSF signaling.Stem Cells. 2005;23:252–263.
Meraz MA, White JM, Sheehan KC, et al. Targeted disruption of the Statl gene in mice reveals unexpected physiologic specificity in the JAK-STAT signaling pathway.Cell. 1996;84:431–442.
Teglund S, McKay C, Schuetz E, et al. Stat5a and Stat5b proteins have essential and nonessential, or redundant, roles in cytokine ingpathway.Cell. 1996;84:431–442.
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Yokoyama, T., Okamura, S., Asano, Y. et al. A Novel Mutation in the Juxtamembrane Intracellular Sequence of the Granulocyte Colony-Stimulating Factor (G-CSF) Receptor Gene in a Patient with Severe Congenital Neutropenia Augments G-CSF Proliferation Activity but Not through the MAP Kinase Cascade. Int J Hematol 82, 28–34 (2005). https://doi.org/10.1532/IJH97.05010
- Congenital neutropenia
- G-CSF receptor
- MAP kinase