Minimal residual disease assessment following CD19‐targeted therapy in B‐cell precursor acute lymphoblastic leukemia using standardized 12‐color flow cytometry: A EuroFlow study
Detection of minimal/measurable residual disease (MRD) is a critical prognostic marker in B-cell precursor acute lymphoblastic leukemia (BCP-ALL). The EuroFlow Consortium previously developed an 8-color flow cytometric MRD protocol, effective for >98% of BCP-ALL patients treated with chemotherapy. This study aimed to enhance MRD detection, particularly for patients treated with CD19-targeted therapies, by expanding the EuroFlow protocol to a 12-color panel. This new panel incorporates additional B-cell markers and exclusion T/NK-cell markers (CD3 and CD7). Through an evaluation of 237 diagnostic BCP-ALL samples, CD22, CD24, and HLA-DR were selected as additional B-cell gating markers. Two 12-color tubes were technically optimized and clinically validated across 101 patient follow-up samples, demonstrating excellent concordance with molecular MRD levels (<i>R</i> <sup>2</sup> = 0.88). The 12-color BCP-ALL MRD tubes were compatible with the previously developed 8-color automated gating and identification (AGI) tool and demonstrated good reproducibility. Our findings indicate that the 12-color panel performs comparably to the 8-color BCP-ALL MRD panel, including both CD19-positive and CD19-negative cases. However, it offers an improved definition of the B-cell lineage, particularly for expert-guided manual data analysis, and provides additional information on the expression of the targetable marker CD22.
Authors
- Romana Jugooa
- Jacques J. M. van Dongen (ORCID: https://orcid.org/0000-0002-3650-7087)
- Robby Engelmann (ORCID: https://orcid.org/0000-0002-1048-3638)
- Beatriz Rodríguez‐Roca (ORCID: https://orcid.org/0000-0003-0113-3318)
- Malicorne Buysse
- Ester Mejstříková (ORCID: https://orcid.org/0000-0001-5169-4653)
- Vincent H. J. van der Velden (ORCID: https://orcid.org/0000-0001-9457-3763)
- Anna Laqua
- Mattias Hofmans (ORCID: https://orcid.org/0000-0003-2061-7617)
- Michaela Reiterová
- Elen Oliveira (ORCID: https://orcid.org/0000-0001-9673-4145)
- Joana Desterro (ORCID: https://orcid.org/0000-0002-6359-7233)
- Łukasz Sędek (ORCID: https://orcid.org/0000-0001-9384-914X)
- Martijn W. C. Verbeek (ORCID: https://orcid.org/0000-0002-7240-5176)
- Chiara Buracchi (ORCID: https://orcid.org/0000-0002-5345-7448)
- Tomasz Szczepański (ORCID: https://orcid.org/0000-0001-5336-261X)
- Saskia Kohlscheen
- Anja X. De Jong
- Monika Brüggemann (ORCID: https://orcid.org/0000-0001-5514-5010)
- Giuseppe Gaipa (ORCID: https://orcid.org/0000-0002-1006-5946)
- Stefan Nierkens (ORCID: https://orcid.org/0000-0003-3406-817X)
- Alberto Órfão (ORCID: https://orcid.org/0000-0002-0007-7230)
- Susana Bárrena
- Sebastian Böettcher
- Elaine Sobral de Costa
- Joana G. Rodriques
Institutions
- Universidade Federal do Rio de Janeiro (BR)
- Medical University of Silesia (PL)
- Universidad de Salamanca (ES)
- Charles University (CZ)
- Leiden University Medical Center (NL)
- Ghent University Hospital (BE)
- Instituto de Salud Carlos III (ES)
- Erasmus MC (NL)
- Christian-Albrechts-Universität zu Kiel (DE)
- Azienda Ospedaliera San Gerardo (IT)
- Princess Máxima Center (NL)
- Instituto de Investigación Biomédica de Salamanca (ES)
- Centro de Investigación del Cáncer (ES)
- Centro de Investigación Biomédica en Red de Cáncer (ES)
- University of Rostock (DE)
Publication Details
- Journal
- HemaSphere
- Published
- 2025-04-01
- DOI
- https://doi.org/10.1002/hem3.70125
- Primary Topic
- Acute Lymphoblastic Leukemia research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Amgen
- Celgene
- Univerzita Karlova v Praze
- Incyte
- European Commission
- Ministerstvo Zdravotnictví Ceské Republiky
- José Carreras Leukämie-Stiftung
- Uniwersytet Śląski w Katowicach
- Instituto de Salud Carlos III
- European Regional Development Fund