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Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling

Persistent measurable residual disease (MRD) is an increasingly important prognostic marker in acute myeloid leukemia (AML). Currently, MRD is determined by multi‐parameter flow cytometry (MFC) or PCR‐based methods detecting leukemia‐specific fusion transcripts and mutations. However, while MFC is h...

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Autores principales: Daga, Shruti, Rosenberger, Angelika, Kashofer, Karl, Heitzer, Ellen, Quehenberger, Franz, Halbwedl, Iris, Graf, Ricarda, Krisper, Nina, Prietl, Barbara, Höfler, Gerald, Reinisch, Andreas, Zebisch, Armin, Sill, Heinz, Wölfler, Albert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7540028/
https://www.ncbi.nlm.nih.gov/pubmed/32602117
http://dx.doi.org/10.1002/ajh.25918
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author Daga, Shruti
Rosenberger, Angelika
Kashofer, Karl
Heitzer, Ellen
Quehenberger, Franz
Halbwedl, Iris
Graf, Ricarda
Krisper, Nina
Prietl, Barbara
Höfler, Gerald
Reinisch, Andreas
Zebisch, Armin
Sill, Heinz
Wölfler, Albert
author_facet Daga, Shruti
Rosenberger, Angelika
Kashofer, Karl
Heitzer, Ellen
Quehenberger, Franz
Halbwedl, Iris
Graf, Ricarda
Krisper, Nina
Prietl, Barbara
Höfler, Gerald
Reinisch, Andreas
Zebisch, Armin
Sill, Heinz
Wölfler, Albert
author_sort Daga, Shruti
collection PubMed
description Persistent measurable residual disease (MRD) is an increasingly important prognostic marker in acute myeloid leukemia (AML). Currently, MRD is determined by multi‐parameter flow cytometry (MFC) or PCR‐based methods detecting leukemia‐specific fusion transcripts and mutations. However, while MFC is highly operator‐dependent and difficult to standardize, PCR‐based methods are only available for a minority of AML patients. Here we describe a novel, highly sensitive and broadly applicable method for MRD detection by combining MFC‐based leukemic cell enrichment using an optimized combinatorial antibody panel targeting CLL‐1, TIM‐3, CD123 and CD117, followed by mutational analysis of recurrently mutated genes in AML. In dilution experiments this method showed a sensitivity of 10(−4) to 10(−5) for residual disease detection. In prospectively collected remission samples this marker combination allowed for a median 67‐fold cell enrichment with sufficient DNA quality for mutational analysis using next generation sequencing (NGS) or digital PCR in 39 out of 41 patients. Twenty‐one samples (53.8%) tested MRD positive, whereas 18 (46.2%) were negative. With a median follow‐up of 559 days, 71.4% of MRD positive (15/21) and 27.8% (5/18) of MRD negative patients relapsed (P = .007). The cumulative incidence of relapse (CIR) was higher for MRD positive patients (5‐year CIR: 90.5% vs 28%, P < .001). In multivariate analysis, MRD positivity was a prominent factor for CIR. Thus, MFC‐based leukemic cell enrichment using antibodies against CLL‐1, TIM‐3, CD123 and CD117 followed by mutational analysis allows high sensitive MRD detection and is informative on relapse risk in the majority of AML patients.
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spelling pubmed-75400282020-10-09 Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling Daga, Shruti Rosenberger, Angelika Kashofer, Karl Heitzer, Ellen Quehenberger, Franz Halbwedl, Iris Graf, Ricarda Krisper, Nina Prietl, Barbara Höfler, Gerald Reinisch, Andreas Zebisch, Armin Sill, Heinz Wölfler, Albert Am J Hematol Research Articles Persistent measurable residual disease (MRD) is an increasingly important prognostic marker in acute myeloid leukemia (AML). Currently, MRD is determined by multi‐parameter flow cytometry (MFC) or PCR‐based methods detecting leukemia‐specific fusion transcripts and mutations. However, while MFC is highly operator‐dependent and difficult to standardize, PCR‐based methods are only available for a minority of AML patients. Here we describe a novel, highly sensitive and broadly applicable method for MRD detection by combining MFC‐based leukemic cell enrichment using an optimized combinatorial antibody panel targeting CLL‐1, TIM‐3, CD123 and CD117, followed by mutational analysis of recurrently mutated genes in AML. In dilution experiments this method showed a sensitivity of 10(−4) to 10(−5) for residual disease detection. In prospectively collected remission samples this marker combination allowed for a median 67‐fold cell enrichment with sufficient DNA quality for mutational analysis using next generation sequencing (NGS) or digital PCR in 39 out of 41 patients. Twenty‐one samples (53.8%) tested MRD positive, whereas 18 (46.2%) were negative. With a median follow‐up of 559 days, 71.4% of MRD positive (15/21) and 27.8% (5/18) of MRD negative patients relapsed (P = .007). The cumulative incidence of relapse (CIR) was higher for MRD positive patients (5‐year CIR: 90.5% vs 28%, P < .001). In multivariate analysis, MRD positivity was a prominent factor for CIR. Thus, MFC‐based leukemic cell enrichment using antibodies against CLL‐1, TIM‐3, CD123 and CD117 followed by mutational analysis allows high sensitive MRD detection and is informative on relapse risk in the majority of AML patients. John Wiley & Sons, Inc. 2020-08-10 2020-10 /pmc/articles/PMC7540028/ /pubmed/32602117 http://dx.doi.org/10.1002/ajh.25918 Text en © 2020 The Authors. American Journal of Hematology published by Wiley Periodicals LLC. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Daga, Shruti
Rosenberger, Angelika
Kashofer, Karl
Heitzer, Ellen
Quehenberger, Franz
Halbwedl, Iris
Graf, Ricarda
Krisper, Nina
Prietl, Barbara
Höfler, Gerald
Reinisch, Andreas
Zebisch, Armin
Sill, Heinz
Wölfler, Albert
Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling
title Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling
title_full Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling
title_fullStr Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling
title_full_unstemmed Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling
title_short Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling
title_sort sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry‐based leukemic cell enrichment followed by mutational profiling
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7540028/
https://www.ncbi.nlm.nih.gov/pubmed/32602117
http://dx.doi.org/10.1002/ajh.25918
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