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The KIT D816V expressed allele burden for diagnosis and disease monitoring of systemic mastocytosis

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Abstract

The activating KIT D816V mutation plays a central role in the pathogenesis, diagnosis, and targeted treatment of systemic mastocytosis (SM). For improved and reliable identification of KIT D816V, we have developed an allele-specific quantitative real-time PCR (RQ-PCR) with an enhanced sensitivity of 0.01–0.1 %, which was superior to denaturing high-performance liquid chromatography (0.5–1 %) or conventional sequencing (10–20 %). Overall, KIT D816 mutations were identified in 146/147 (99 %) of patients (D816V, n = 142; D816H, n = 2; D816Y, n = 2) with SM, including indolent SM (ISM, n = 63, 43 %), smoldering SM (n = 8, 5 %), SM with associated hematological non-mast cell lineage disease (SM-AHNMD, n = 16, 11 %), and aggressive SM/mast cell leukemia ± AHNMD (ASM/MCL, n = 60, 41 %). If positive in BM, the KIT D816V mutation was found in PB of all patients with advanced SM (SM-AHNMD, ASM, and MCL) and in 46 % (23/50) of patients with ISM. There was a strong correlation between the KIT D816V expressed allele burden (KIT D816V EAB) with results obtained from DNA by genomic allele-specific PCR and also with disease activity (e.g., serum tryptase level), disease subtype (e.g., indolent vs. advanced SM) and survival. In terms of monitoring of residual disease, qualitative and quantitative assessment of KIT D816V and KIT D816V EAB was successfully used for sequential analysis after chemotherapy or allogeneic stem cell transplantation. We therefore conclude that RQ-PCR assays for KIT D816V are useful complimentary tools for diagnosis, disease monitoring, and evaluation of prognosis in patients with SM.

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Acknowledgments

This work was supported by the ‘Deutsche José Carreras Leukämie-Stiftung e.V.’ (grant no. R09/29f and H11/03).

Authorship

PE, JS, MJ, SS, TE, MM, AF and MG performed the laboratory work for the study; AH, AR, NCPC, KH and WKH provided patient material; JS, MT, GM and AR collected patient information; HPH, AM and KS reviewed the bone marrow biopsies; AR and NCP prepared the study design; JS, PE and AR wrote the paper; PE and JS performed the statistical analyses; AH, WKH and NCPC revised the manuscript; all authors approved the final version of the manuscript.

Conflict-of-interest disclosure

The authors declare no competing financial interests.

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Authors and Affiliations

  1. III. Medizinische Klinik, Universitätsmedizin Mannheim, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany

    Philipp Erben, Juliana Schwaab, Georgia Metzgeroth, Mohamad Jawhar, Alice Fabarius, Martina Teichmann, Martin C. Müller, Michelle Giehl, Wolf-Karsten Hofmann & Andreas Reiter

  2. Pathologisches Institut, Ludwig-Maximilians-Universität München, München, Germany

    Hans-Peter Horny & Karl Sotlar

  3. Institut für Klinische Chemie, Universitätsmedizin Mannheim, Mannheim, Germany

    Sven Schneider

  4. Abteilung Hämatologie/Onkologie, Klinik für Innere Medizin II, Universitätsklinikum Jena, Jena, Germany

    Thomas Ernst & Andreas Hochhaus

  5. Pathologisches Institut, Universitätsmedizin Mannheim, 68167, Mannheim, Germany

    Alexander Marx

  6. Klinik für Dermatologie, Universitätsklinik Köln, Köln, Germany

    Karin Hartmann

  7. Wessex Regional Genetics Laboratory, Salisbury, UK

    Nicholas C. P. Cross

  8. Faculty of Medicine, University of Southampton, Southampton, UK

    Nicholas C. P. Cross

Authors
  1. Philipp Erben
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  18. Andreas Reiter
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Corresponding author

Correspondence to Andreas Reiter.

Additional information

P. Erben and J. Schwaab both authors contributed equally to this work

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Erben, P., Schwaab, J., Metzgeroth, G. et al. The KIT D816V expressed allele burden for diagnosis and disease monitoring of systemic mastocytosis. Ann Hematol 93, 81–88 (2014). https://doi.org/10.1007/s00277-013-1964-1

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  • DOI: https://doi.org/10.1007/s00277-013-1964-1

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