Abstract
Purpose
We report normal neutrophil count in a mother, who carries the same ELANE mutation as her daughter with severe congenital neutropenia. We hypothesized that the mother possessed wild- and mutant-type clones and the wild-type clones could generate neutrophils, whereas the mutant clones could not.
Methods
We confirmed mutant variant ratio by sequence signals and measured the frequency of the mutant allele by subcloning in various cell types. We established the ELANE-mutated and non-mutated induced pluripotent stem cells (iPSCs) from the mother’s T cells and compared granulopoiesis between these iPSCs.
Results
In the sequence analysis of isolated peripheral blood (PB), nail and hair, the mutant variant was detected in approximately 40–60% of lymphocytes, monocytes, hematopoietic progenitor cells, and hair as well as in a small percentage of nail, but in none of the neutrophils. In the subcloning analysis of extracted DNA from CD3+ and CD34+ cells, the mutant allele was identified in 37.5% and 38.1%, respectively. We reprogrammed the mother’s PB cells and established the ELANE-mutated and non-mutated iPSCs. Granulopoiesis from mutated iPSCs revealed little sensitivity to granulocyte colony-stimulating factor in comparison with non-mutated iPSCs.
Conclusions
These observations strongly suggest that mutant-carrying neutrophils did not appear in the mother’s PB because mutated clones could not differentiate into neutrophils. The mother’s normal hematological phenotype could be explained by the perseverance of normal, non-mutated granulopoiesis.
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References
Welte K, Dale D. Pathophysiology and treatment of severe chronic neutropenia. Ann Hematol. 1996;72:158–65.
Dale DC, Person RE, Bolyard AA, Aprikyan AG, Bos C, Bonilla MA, et al. Mutations in the gene encoding neutrophil elastase in congenital and cyclic neutropenia. Blood. 2000;96:2317–22.
Boxer LA, Newburger PE. A molecular classification of congenital neutropenia syndromes. Pediatr Blood Cancer. 2007;49:609–14.
Bellanne-Chantelot C, Clauin S, Leblanc T, Cassinat B, Rodrigues-Lima F, Beaufils S, et al. Mutations in the ELA2 gene correlate with more severe expression of neutropenia: a study of 81 patients from the French neutropenia register. Blood. 2004;103:4119–25.
Rosenberg PS, Alter BP, Link DC, Stein S, Rodger E, Bolyard AA, et al. Neutrophil elastase mutations and risk of leukaemia in severe congenital neutropenia. Br J Haematol. 2008;140:210–3.
Briars GL, Parry HF, Ansari BM. Dominantly inherited severe congenital neutropenia. J Inf Secur. 1996;33:123–6.
Boxer LA, Stein S, Buckley D, Bolyard AA, Dale DC. Strong evidence for autosomal dominant inheritance of severe congenital neutropenia associated with ELA2 mutations. J Pediatr. 2006;148:633–6.
Kollner I, Sodeik B, Schreek S, Heyn H, von Neuhoff N, Germeshausen M, et al. Mutations in neutrophil elastase causing congenital neutropenia lead to cytoplasmic protein accumulation and induction of the unfolded protein response. Blood. 2006;108:493–500.
Tidwell T, Wechsler J, Nayak RC, Trump L, Salipante SJ, Cheng JC, et al. Neutropenia-associated ELANE mutations disrupting translation initiation produce novel neutrophil elastase isoforms. Blood. 2014;123:562–9.
Newburger PE, Pindyck TN, Zhu Z, Bolyard AA, Aprikyan AAG, Dale DC, et al. Cyclic neutropenia and severe congenital neutropenia in patients with a shared ELANE mutation and paternal haplotype: evidence for phenotype determination by modifying genes. Pediatr Blood Cancer. 2010;55:314–7.
Sakashita K, Kato I, Daifu T, Saida S, Hiramatsu H, Nishinaka Y, et al. In vitro expansion of CD34(+)CD38(−) cells under stimulation with hematopoietic growth factors on AGM-S3 cells in juvenile myelomonocytic leukemia. Leukemia. 2015;29:606–14.
Seki T, Yuasa S, Oda M, Egashira T, Yae K, Kusumoto D, et al. Generation of induced pluripotent stem cells from human terminally differentiated circulating T cells. Cell Stem Cell. 2010;7:11–4.
Ma F, Wang D, Hanada S, Ebihara Y, Kawasaki H, Zaike Y, et al. Novel method for efficient production of multipotential hematopoietic progenitors from human embryonic stem cells. Int J Hematol. 2007;85:371–9.
Hiramoto T, Ebihara Y, Mizoguchi Y, Nakamura K, Yamaguchi K, Ueno K, et al. Wnt3a stimulates maturation of impaired neutrophils developed from severe congenital neutropenia patient-derived pluripotent stem cells. Proc Natl Acad Sci U S A. 2013;110:3023–8.
Nayak RC, Trump LR, Aronow BJ, Myers K, Mehta P, Kalfa T, et al. Pathogenesis of ELANE-mutant severe neutropenia revealed by induced pluripotent stem cells. J Clin Invest. 2015;125:3103–16.
Kanda Y. Investigation of the freely available easy-to-use software 'EZR' for medical statistics. Bone Marrow Transplant. 2013;48:452–8.
Xia J, Bolyard AA, Rodger E, Stein S, Aprikyan AA, Dale DC, et al. Prevalence of mutations in ELANE, GFI1, HAX1, SBDS, WAS and G6PC3 in patients with severe congenital neutropenia. Br J Haematol. 2009;147:535–42.
Shim YJ, Kim HJ, Suh JS, Lee KS. Novel ELANE gene mutation in a Korean girl with severe congenital neutropenia. J Korean Med Sci. 2011;26:1646–9.
Germeshausen M, Ballmaier M, Welte K. Incidence of CSF3R mutations in severe congenital neutropenia and relevance for leukemogenesis: results of a long-term survey. Blood. 2007;109:93–9.
Germeshausen M, Schulze H, Ballmaier M, Zeidler C, Welte K. Mutations in the gene encoding neutrophil elastase (ELA2) are not sufficient to cause the phenotype of congenital neutropenia. Br J Haematol. 2001;115:222–4.
Ancliff PJ, Gale RE, Watts MJ, Liesner R, Hann IM, Strobel S, et al. Paternal mosaicism proves the pathogenic nature of mutations in neutrophil elastase in severe congenital neutropenia. Blood. 2002;100:707–9.
Kim HJ, Song MJ, Lee KO, Kim SH, Kim HJ. Paternal somatic mosaicism of a novel frameshift mutation in ELANE causing severe congenital neutropenia. Pediatr Blood Cancer. 2015;62:2229–31.
Hirata O, Okada S, Tsumura M, Karakawa S, Matsumura I, Kimura Y, et al. Mosaicism of an ELANE mutation in an asymptomatic mother in a familial case of cyclic neutropenia. J Clin Immunol. 2015;35:512–6.
Acknowledgments
The authors would like to thank Enago (www.enago.jp) for the English language review and Yūka Miyajima for assistance in preparing the manuscript.
Funding
This work was supported by the Japan Society Promotion of Science KAKENHI Grant Number 17K10104.
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T.S. and O.O. performed experiments; T.S., N.K., and K.A. treated the patient; and T.S. and Y.N. designed the study and drafted the manuscript.
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This study was performed following approval from the independent ethics committee of Shinshu University School of Medicine, Matsumoto, Japan; written informed consent was obtained from participants.
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The authors declare that they have no competing interests.
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Shigemura, T., Kobayashi, N., Agematsu, K. et al. Mosaicism of an ELANE Mutation in an Asymptomatic Mother. J Clin Immunol 39, 106–111 (2019). https://doi.org/10.1007/s10875-018-0580-1
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DOI: https://doi.org/10.1007/s10875-018-0580-1