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Mosaicism of an ELANE Mutation in an Asymptomatic Mother in a Familial Case of Cyclic Neutropenia

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Abstract

Purpose

To confirm and characterize mosaicism of the cyclic neutropenia (CyN)-related mutation in the ELANE gene identified in the asymptomatic mother of patients with CyN.

Methods

We identified sibling cases with CyN due to a novel heterozygous splicing site mutation, IVS4 +5SD G>T, in the ELANE gene, resulting in an internal in-frame deletion of 30 nucleotides (corresponding to a ten amino acid deletion, V161–F170). The mutated allele was also detected in their asymptomatic mother but at low frequency. We measured the frequency of the mutant allele from peripheral blood leukocytes (PBLs) by subcloning, and confirmed the allelic frequency of mosaicism in various cell types by massively parallel DNA sequencing (MPS) analysis.

Results

In the subcloning analysis, the mutant allele was identified in 21.36 % of PBLs from the asymptomatic mother, compared with 54.72 % of PBLs from the CyN patient. In the MPS analysis, the mutant allele was observed in approximately 30 % of mononuclear cells, CD3+ T cells, CD14+ monocytes and the buccal mucosa. Conversely, it was detected in low frequency in polymorphonuclear leukocytes (PLMLs) (3–4 %) and CD16+ granulocytes (2–3 %).

Conclusions

Mosaicism of the ELANE mutation has only previously been identified in one confirmed and one unconfirmed case of SCN. This is the first report of mosaicism of the ELANE mutation in a case of CyN. The MPS results suggest that this de novo mutation occurred during the two-cell stage of embryogenesis. PLMLs expressing the ELANE mutation were found to be actively undergoing apoptosis.

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References

  1. Dale DC, Hammond WP. Cyclic neutropenia: a clinical review. Blood Rev. 1988;2:178–85.

    Article  CAS  PubMed  Google Scholar 

  2. Palmer SE, Stephens K, Dale DC. Genetics, phenotype, and natural history of autosomal dominant cyclic hematopoiesis. Am J Med Genet. 1996;66:413–22.

    Article  CAS  PubMed  Google Scholar 

  3. 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.

    CAS  PubMed  Google Scholar 

  4. 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;23:433–6.

    Article  CAS  PubMed  Google Scholar 

  5. Horwitz MS, Duan Z, Korkmaz B, Lee HH, Mealiffe ME, Salipante SJ. Neutrophil elastase in cyclic and severe congenital neutropenia. Blood. 2007;109:1817–24.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. 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.

    Article  CAS  PubMed  Google Scholar 

  7. Schäffer AA, Klein C. Genetic heterogeneity in severe congenital neutropenia: how many aberrant pathways can kill a neutrophil? Curr Opin Allergy Clin Immunol. 2007;7:481–94.

    Article  PubMed Central  PubMed  Google Scholar 

  8. Ishikawa N, Okada S, Miki M, Shirao K, Kihara H, Tsumura M, et al. Neurodevelopmental abnormalities associated with severe congenital neutropenia due to the R86X mutation in the HAX1 gene. J Med Genet. 2008;45:802–7.

    Article  CAS  PubMed  Google Scholar 

  9. Sera Y, Kawaguchi H, Nakamura K, Sato T, Habara M, Okada S, et al. A comparison of the defective granulopoiesis in childhood cyclic neutropenia and in severe congenital neutropenia. Haematologica. 2005;90:1032–41.

    CAS  PubMed  Google Scholar 

  10. Welte K, Zeidler C, Dale DC. Severe congenital neutropenia. Semin Hematol. 2006;43:189–95.

    Article  CAS  PubMed  Google Scholar 

  11. 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.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Izawa K, Hijikata A, Tanaka N, Kawai T, Saito MK, Goldbach-Mansky R, et al. Detection of base substitution-type somatic mosaicism of the NLRP3 gene with >99.9 % statistical confidence by massively parallel sequencing. DNA Res. 2012;19:143–52.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. Flaherty P, Natsoulis G, Muralidharan O, Winters M, Buenrostro J, Bell J, et al. Ultrasensitive detection of rare mutations using next-generation targeted resequencing. Nucleic Acids Res. 2012;40:e2.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. 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.

    Article  CAS  PubMed  Google Scholar 

  15. Benson KF, Horwitz M. Possibility of somatic mosaicism of ELA2 mutation overlooked in an asymptomatic father transmitting severe congenital neutropenia to two offspring. Br J Haematol. 2002;118:923. author reply923–4.

    PubMed  Google Scholar 

  16. 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.

    Article  CAS  PubMed  Google Scholar 

  17. Grenda DS, Murakami M, Ghatak J, Xia J, Boxer LA, Dale D, et al. Mutations of the ELA2 gene found in patients with severe congenital neutropenia induce the unfolded protein response and cellular apoptosis. Blood. 2007;110:4179–87.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Klein C. Genetic defects in severe congenital neutropenia: emerging insights into life and death of human neutrophil granulocytes. Annu Rev Immunol. 2011;29:399–413.

    Article  CAS  PubMed  Google Scholar 

  19. Köllner 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.

    Article  PubMed  Google Scholar 

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Acknowledgments

This study was supported in part by Grants in Aid for Scientific Research from the Japan Society for the Promotion of Science [22591161 to M.K.], [25713039 to S.O.] and [80457241 to S.O.]. This study was also supported in part by Research on Measures for Intractable Diseases funding from the Japanese Ministry of Health, Labour and Welfare [H22-Nanchi-ippan-078 to M.K.]. Sequence analysis was supported by the Analysis Center of Life Science, Natural Science Center for Basic Research and Development, Hiroshima University (Hiroshima, Japan).

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Authors and Affiliations

  1. Department of Pediatrics, Hiroshima University Graduate School of Biomedical & Health Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Osamu Hirata, Satoshi Okada, Miyuki Tsumura, Shuhei Karakawa & Masao Kobayashi

  2. Department of Hematology and Rheumatology, Kinki University Faculty of Medicine, Osaka, Japan

    Itaru Matsumura

  3. Department of Pediatrics, Kyotominami Hospital, Kyoto, Japan

    Yujiro Kimura

  4. Department of Pediatrics, Hyogo Prefectural Tsukaguchi Hospital, Hyogo, Japan

    Toshiro Maihara

  5. Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima, Japan

    Shin’ichiro Yasunaga & Yoshihiro Takihara

  6. Department of Technology Development, Kazusa DNA Research Institute, Chiba, Japan

    Osamu Ohara

Authors
  1. Osamu Hirata
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  2. Satoshi Okada
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  3. Miyuki Tsumura
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  4. Shuhei Karakawa
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  5. Itaru Matsumura
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  6. Yujiro Kimura
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  7. Toshiro Maihara
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  8. Shin’ichiro Yasunaga
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  9. Yoshihiro Takihara
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  10. Osamu Ohara
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  11. Masao Kobayashi
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Corresponding author

Correspondence to Masao Kobayashi.

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Hirata, O., Okada, S., Tsumura, M. et al. Mosaicism of an ELANE Mutation in an Asymptomatic Mother in a Familial Case of Cyclic Neutropenia. J Clin Immunol 35, 512–516 (2015). https://doi.org/10.1007/s10875-015-0165-1

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  • DOI: https://doi.org/10.1007/s10875-015-0165-1

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