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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

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1437-4331
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Volume 57, Issue 5

Issues

Droplet digital PCR for the simultaneous analysis of minimal residual disease and hematopoietic chimerism after allogeneic cell transplantation

Miguel Waterhouse
  • Corresponding author
  • Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
  • Core Facility, Department of Hematology, Oncology and Stem cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
  • Molecular Diagnostics Lab, Department of Hematology, Oncology and Stem cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
  • Department of Hematology/Oncology, University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany
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/ Dietmar Pfeifer
  • Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
  • Core Facility, Department of Hematology, Oncology and Stem cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
  • Molecular Diagnostics Lab, Department of Hematology, Oncology and Stem cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
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/ Jesus Duque-Afonso
  • Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
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/ Marie Follo
  • Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
  • Core Facility, Department of Hematology, Oncology and Stem cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
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/ Justus Duyster
  • Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
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/ Melanie Depner
  • Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
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/ Hartmut Bertz
  • Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
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/ Jürgen Finke
  • Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany
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Published Online: 2018-11-20 | DOI: https://doi.org/10.1515/cclm-2018-0827

Abstract

Background

Minimal residual disease (MRD) and hematopoietic chimerism testing influences clinical decision and therapeutic intervention in patients after allogeneic stem cell transplantation (HSCT). However, treatment approaches to induce complete donor chimerism and MRD negativity can lead to complications such as graft-versus-host disease (GvHD) and marrow aplasia. Therefore, there is a need for comprehensive characterization of the molecular remission status after transplantation.

Methods

We analyzed 764 samples from 70 patients after HSCT for the simultaneous measurement of chimerism and molecular targets used for MRD testing with a digital PCR (dPCR) platform.

Results

Mixed chimerism (MC) was detected in 219 samples from 37 patients. The mean percentage of host derived DNA in these clinical samples was 4.3%. Molecular relapse with a positive MRD marker and/or increased WT1 expression was observed in 15 patients. In addition to WT1 overexpression, other MRD positive markers were: NPM1 (Type A, B, K), DNMT3A (R882H), MLL-PTD, IDH1 (R132H) and KRAS (G12S). Increasing MC was observed in 15 patients. This group of patients showed either a positive MRD marker, increased WT1 expression or both. Next, we analyzed whether MC or the molecular target for MRD was first detected. MC and MRD marker positivity in this group was first detected in six and two patients, respectively. In the remaining seven patients MC and MRD positivity was detected simultaneously.

Conclusions

The combination of MRD and chimerism markers in a dPCR platform represents a practical, sensitive and accurate diagnostic tool for the comprehensive assessment of the molecular remission status of patients undergoing HSCT.

Keywords: allogeneic hematopoietic cell transplantation; chimerism; droplet digital PCR; minimal residual disease

References

  • 1.

    Jacobsohn DA, Loken MR, Fei M, Adams A, Brodersen LE, Logan BR, et al. Outcomes of measurable residual disease in pediatric acute myeloid leukemia pre- and post-hematopoietic stem cell transplant: validation of difference from normal flow cytometry with chimerism studies and Wilms tumor 1 gene expression. Biol Blood Marrow Transplant 2018 Jun 19. pii: S1083-8791(18)30322-7. doi: 10.1016/j.bbmt.2018.06.010. [Epub ahead of print].Google Scholar

  • 2.

    Araki D, Wood BL, Othus M, Radich JP, Halpern AB, Zhou Y, et al. Allogeneic hematopoietic cell transplantation for acute myeloid leukemia: time to move toward a minimal residual disease-based definition of complete remission? J Clin Oncol 2016;34:329–36.CrossrefWeb of ScienceGoogle Scholar

  • 3.

    Choi SJ, Lee KH, Lee JH, Kim S, Chung HJ, Lee JS, et al. Pronogstic value of hematopoietic chimerism in patients with acute leukemia after allogeneic bone marrow transplantation: a prospective study. Bone Marrow Transplant 2000;26:327–32.CrossrefGoogle Scholar

  • 4.

    Kreyenberg H, Hölle W, Möhrle S, Niethammer D, Bader P. Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Tuebingen experience. Leukemia 2003;17:237–40.CrossrefPubMedGoogle Scholar

  • 5.

    Alizadeh M, Bernard M, Danic B, Dauriac C, Birebent B, Lapart C, et al. Quantitative assessment of hematopoietic chimerism after bone marrow transplantation by real-time quantitative polymerase chain reaction. Blood 2002;99:4618–25.CrossrefPubMedGoogle Scholar

  • 6.

    Waterhouse M, Pfeifer D, Follo M, Duyster J, Schäfer H, Bertz H, et al. Early mixed hematopoietic chimerism detection by digital droplet PCR in patients undergoing gender-mismatched hematopoietic stem cell transplantation. Clin Chem Lab Med 2017;55:1115–21.CrossrefWeb of SciencePubMedGoogle Scholar

  • 7.

    Aloisio M, Licastro D, Caenazzo L, Torboli V, D’Eustacchio A, Severini GM, et al. A technical application of quantitative next generation sequencing for chimerism evaluation. Mol Med Rep 2016;14:2967–74.CrossrefWeb of SciencePubMedGoogle Scholar

  • 8.

    Lee HC, Saliba RM, Rondon G, Chen J, Charafeddine Y, Medeiros LJ, et al. Mixed T lymphocyte chimerism after allogeneic hematopoietic transplantation is predictive for relapse of acute myeloid leukemia and myelodysplastic syndromes. Biol Blood Marrow Transplant 2015;21:1948–54.Web of ScienceCrossrefPubMedGoogle Scholar

  • 9.

    Terwey TH, Hemmati PG, Nagy M, Pfeifer H, Gökbuget N, Brüggemann M, et al. Comparison of chimerism and minimal residual disease monitoring for relapse prediction after allogeneic stem cell transplantation for acute lymphoblastic leukemia. Biol Blood Marrow Transplant 2014;20:1522–9.CrossrefWeb of SciencePubMedGoogle Scholar

  • 10.

    Zeiser R, Spyridonidis A, Wäsch R, Ihorst G, Grüllich C, Bertz H, et al. Evaluation of immunomodulatory treatment based on conventional and lineage-specific chimerism analysis in patients with myeloid malignancies after myeloablative allogeneic hematopoietic cell transplantation. Leukemia 2005;19:814–21.CrossrefPubMedGoogle Scholar

  • 11.

    Stahl T, Böhme MU, Kröger N, Fehse B. Digital PCR to assess hematopoietic chimerism after allogeneic stem cell transplantation. Exp Hematol 2015;43:462–8.CrossrefWeb of SciencePubMedGoogle Scholar

  • 12.

    George D, Czech J, John B, Yu M, Jennings LJ. Detection and quantification of chimerism by droplet digital PCR. Chimerism 2013;4:102–8.PubMedCrossrefGoogle Scholar

  • 13.

    Sellmann L, Rabe K, Bünting I, Dammann E, Göhring G, Ganser A, et al. Diagnostic value of highly-sensitive chimerism analysis after allogeneic stem cell transplantation. Bone Marrow Transplant 2018 May 2. doi: 10.1038/s41409-018-0176-7. [Epub ahead of print].PubMedWeb of ScienceGoogle Scholar

  • 14.

    Marks R, Potthoff K, Hahn J, Ihorst G, Bertz H, Spyridonidis A, et al. Reduced-toxicity conditioning with fludarabine, BCNU and melphalan in allogeneic cell transplantation: particular activity against advanced hematologic malignancies. Blood 2008;112:415–25.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 15.

    Clark JR, Scott SD, Jack AL, Lee H, Mason J, Carter GI, et al. Monitoring of chimerism following allogeneic haematopoietic stem cell transplantation (HSCT): technical recommendations for the use of short tandem repeats (STR) based techniques, on behalf of the United Kingdom National External Quality Assessment service for Leucocyte Immunophenotyping Chimerism Working Group. Br J Haematol 2015;168:26–37.Google Scholar

  • 16.

    Clinical and Laboratory Standards Institute. EP17-A2 Evaluation of detection capability for Clinical Laboratory Measurement Procedures; Approved Guideline-Second Edition. Wayne, PA: CLSI;2013.Google Scholar

  • 17.

    Cilloni D, Renneville A, Hermitte F, Hills RK, Daly S, Jovanovic JV, et al. Real-time quantitative polymerase chain reaction detection of minimal residual disease by standardized WT1 assay to enhance risk stratification in acute myeloid leukemia: a European LeukemiaNet study. J Clin Oncol 2009;27:5195–201.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 18.

    Frey NV, Porter DL. Graft-versus-host disease after donor leukocyte infusions: presentation and management. Best Pract Res Clin Haematol 2008;38:399–405.Web of ScienceGoogle Scholar

  • 19.

    Garicochea B, van Rhee F, Spencer A, Chase A, Lin F, Cross NC, et al. Aplasia after donor lymphocyte infusion (DLI) for CML in relapse after sex-mismatched BMT: recovery of donor-type haemopoiesis predicted by non-isotopic in situ hybridization (ISH). Br J Haematol 1994;88:400–2.CrossrefPubMedGoogle Scholar

  • 20.

    Waterhouse M, Follo M, Pfeifer D, von Bubnoff N, Duyster J, Bertz H, et al. Sensitive and accurate quantification of JAK2 V617F mutation in chronic myeloproliferative neoplasms by droplet digital PCR. Ann Hematol 2016;95:739–44.Web of SciencePubMedCrossrefGoogle Scholar

  • 21.

    Ommen HB, Hokland P, Haferlach T, Abildgaard L, Alpermann T, Haferlach C, et al. Relapse kinetics in acute myeloid leukemias with MLL translocations or partial tandem duplications within the MLL gene. Br J Haematol 2014;165:618–28.CrossrefPubMedGoogle Scholar

  • 22.

    Papadaki C, Dufour A, Seibl M, Schneider S, Bohlander SK, Zellmeier E, et al. Monitoring minimal residual disease in acute myeloid leukaemia with NPM1 mutations by quantitative PCR: clonal evolution is a limiting factor. Br J Haematol 2009;144:517–23.Web of SciencePubMedCrossrefGoogle Scholar

About the article

Corresponding author: Miguel Waterhouse, PhD, Department of Hematology, Oncology and Stem Cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany; Core Facility, Department of Hematology, Oncology and Stem cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany; Molecular Diagnostics Lab, Department of Hematology, Oncology and Stem cell Transplantation, University of Freiburg, Faculty of Medicine, Freiburg, Germany; and Department of Hematology/Oncology, University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany, Phone: +49 761 270 3646, Fax: +49 761 270 3582


Received: 2018-08-02

Accepted: 2018-10-25

Published Online: 2018-11-20

Published in Print: 2019-04-24


Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 57, Issue 5, Pages 641–647, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2018-0827.

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