Technical Aspects of Flow Cytometry-based Measurable Residual Disease Quantification in Acute Myeloid Leukemia: Experience of the European LeukemiaNet MRD Working Party

Overview

Journal Hemasphere

Publisher Wiley

Specialty Hematology

Date 2021 Dec 29

PMID 34964040

Citations 27

Authors

Jesse M Tettero

Sylvie Freeman

Veit Buecklein

Adriano Venditti

Luca Maurillo

Wolfgang Kern

Roland B Walter

Brent L Wood

Christophe Roumier

Jan Philippe

Barbara Denys

Jeffrey L Jorgensen

Marie C Bene

Francis Lacombe

Adriana Plesa

Monica L Guzman

Agnieszka Wierzbowska

Anna Czyz

Lok Lam Ngai

Adrian Schwarzer

Costa Bachas

Jacqueline Cloos

Marion Subklewe

Michaela Fuering-Buske

Francesco Buccisano

Affiliations

Soon will be listed here.

Abstract

Measurable residual disease (MRD) quantified by multiparameter flow cytometry (MFC) is a strong and independent prognostic factor in acute myeloid leukemia (AML). However, several technical factors may affect the final read-out of the assay. Experts from the MRD Working Party of the European LeukemiaNet evaluated which aspects are crucial for accurate MFC-MRD measurement. Here, we report on the agreement, obtained via a combination of a cross-sectional questionnaire, live discussions, and a Delphi poll. The recommendations consist of several key issues from bone marrow sampling to final laboratory reporting to ensure quality and reproducibility of results. Furthermore, the experiences were tested by comparing two 8-color MRD panels in multiple laboratories. The results presented here underscore the feasibility and the utility of a harmonized theoretical and practical MFC-MRD assessment and are a next step toward further harmonization.

Citing Articles

Impact of high-sensitivity flow cytometry on peri-transplant minimal residual disease kinetics in acute leukemia.

de Azambuja A, Beltrame M, Malvezzi M, Schluga Y, Justus J, Lima A Sci Rep. 2025; 15(1):6942.

PMID: 40011589 PMC: 11865467. DOI: 10.1038/s41598-025-91936-7.

Significance of Measurable Residual Disease in Patients Undergoing Allogeneic Hematopoietic Cell Transplantation for Acute Myeloid Leukemia.

Gang M, Othus M, Walter R Cells. 2025; 14(4).

PMID: 39996762 PMC: 11853423. DOI: 10.3390/cells14040290.

Fludarabine melphalan reduced intensity conditioning vs radiation-based myeloablative conditioning in patients undergoing allogeneic transplantation for acute myeloid leukemia with measurable residual disease.

Blackmon A, Afkhami M, Yang D, Mokhtari S, Samara Y, Pourhassan H Bone Marrow Transplant. 2024; 60(2):165-174.

PMID: 39695333 PMC: 11810767. DOI: 10.1038/s41409-024-02491-0.

Impact of measurable residual disease clearance kinetics in patients with AML undergoing intensive chemotherapy.

Jen W, Jen W, Sasaki K, Ravandi F, Kadia T, Wang S Blood Adv. 2024; 9(4):783-792.

PMID: 39631072 PMC: 11869955. DOI: 10.1182/bloodadvances.2024013826.

Detection methods and prognosis implications of measurable residual disease in acute myeloid leukemia.

Zhao Z, Lan J Ann Hematol. 2024; 103(12):4869-4881.

PMID: 39283479 DOI: 10.1007/s00277-024-06008-z.

References

Jongen-Lavrencic M, Grob T, Hanekamp D, Kavelaars F, Al Hinai A, Zeilemaker A . Molecular Minimal Residual Disease in Acute Myeloid Leukemia. N Engl J Med. 2018; 378(13):1189-1199. DOI: 10.1056/NEJMoa1716863. View

Jawhar M, Dohner K, Kreil S, Schwaab J, Shoumariyeh K, Meggendorfer M . KIT D816 mutated/CBF-negative acute myeloid leukemia: a poor-risk subtype associated with systemic mastocytosis. Leukemia. 2019; 33(5):1124-1134. PMC: 6756067. DOI: 10.1038/s41375-018-0346-z. View

Holdrinet R, von Egmond J, Wessels J, Haanen C . A method for quantification of peripheral blood admixture in bone marrow aspirates. Exp Hematol. 1980; 8(1):103-7. View

Craddock C, Jackson A, Loke J, Siddique S, Hodgkinson A, Mason J . Augmented Reduced-Intensity Regimen Does Not Improve Postallogeneic Transplant Outcomes in Acute Myeloid Leukemia. J Clin Oncol. 2020; 39(7):768-778. PMC: 8078252. DOI: 10.1200/JCO.20.02308. View

Flores-Montero J, Sanoja-Flores L, Paiva B, Puig N, Garcia-Sanchez O, Bottcher S . Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma. Leukemia. 2017; 31(10):2094-2103. PMC: 5629369. DOI: 10.1038/leu.2017.29. View

Elghetany M, Davis B . Impact of preanalytical variables on granulocytic surface antigen expression: a review. Cytometry B Clin Cytom. 2005; 65(1):1-5. DOI: 10.1002/cyto.b.20051. View

Tomlinson B, Lazarus H . Enhancing acute myeloid leukemia therapy - monitoring response using residual disease testing as a guide to therapeutic decision-making. Expert Rev Hematol. 2017; 10(6):563-574. DOI: 10.1080/17474086.2017.1326811. View

Hanekamp D, Bachas C, van de Loosdrecht A, Ossenkoppele G, Cloos J . Re: Myeloblasts in normal bone marrows expressing leukaemia-associated immunophenotypes. Pathology. 2019; 52(2):289-291. DOI: 10.1016/j.pathol.2019.09.021. View

Westers T, Ireland R, Kern W, Alhan C, Balleisen J, Bettelheim P . Standardization of flow cytometry in myelodysplastic syndromes: a report from an international consortium and the European LeukemiaNet Working Group. Leukemia. 2012; 26(7):1730-41. DOI: 10.1038/leu.2012.30. View

10.

Jaiswal S, Fontanillas P, Flannick J, Manning A, Grauman P, Mar B . Age-related clonal hematopoiesis associated with adverse outcomes. N Engl J Med. 2014; 371(26):2488-98. PMC: 4306669. DOI: 10.1056/NEJMoa1408617. View

11.

Armbruster D, Pry T . Limit of blank, limit of detection and limit of quantitation. Clin Biochem Rev. 2008; 29 Suppl 1:S49-52. PMC: 2556583. View

12.

Hasserjian R . Flow cytometry reveals the nuances of clonal haematopoiesis. Br J Haematol. 2021; 192(6):949-950. DOI: 10.1111/bjh.17348. View

13.

Wood B, Jevremovic D, Bene M, Yan M, Jacobs P, Litwin V . Validation of cell-based fluorescence assays: practice guidelines from the ICSH and ICCS - part V - assay performance criteria. Cytometry B Clin Cytom. 2013; 84(5):315-23. DOI: 10.1002/cyto.b.21108. View

14.

Zeijlemaker W, Kelder A, Oussoren-Brockhoff Y, Scholten W, Snel A, Veldhuizen D . A simple one-tube assay for immunophenotypical quantification of leukemic stem cells in acute myeloid leukemia. Leukemia. 2015; 30(2):439-46. DOI: 10.1038/leu.2015.252. View

15.

Monaco G, Chen H, Poidinger M, Chen J, de Magalhaes J, Larbi A . flowAI: automatic and interactive anomaly discerning tools for flow cytometry data. Bioinformatics. 2016; 32(16):2473-80. DOI: 10.1093/bioinformatics/btw191. View

16.

Buccisano F, Maurillo L, Gattei V, Del Poeta G, Del Principe M, Cox M . The kinetics of reduction of minimal residual disease impacts on duration of response and survival of patients with acute myeloid leukemia. Leukemia. 2006; 20(10):1783-9. DOI: 10.1038/sj.leu.2404313. View

17.

Buckley S, Wood B, Othus M, Hourigan C, Ustun C, Linden M . Minimal residual disease prior to allogeneic hematopoietic cell transplantation in acute myeloid leukemia: a meta-analysis. Haematologica. 2017; 102(5):865-873. PMC: 5477605. DOI: 10.3324/haematol.2016.159343. View

18.

Wood B . Principles of minimal residual disease detection for hematopoietic neoplasms by flow cytometry. Cytometry B Clin Cytom. 2015; 90(1):47-53. DOI: 10.1002/cyto.b.21239. View

19.

Hourigan C, Gale R, Gormley N, Ossenkoppele G, Walter R . Measurable residual disease testing in acute myeloid leukaemia. Leukemia. 2017; 31(7):1482-1490. DOI: 10.1038/leu.2017.113. View

20.

Delgado J, Guillen-Grima F, Moreno C, Panizo C, Perez-Robles C, Mata J . A simple flow-cytometry method to evaluate peripheral blood contamination of bone marrow aspirates. J Immunol Methods. 2017; 442:54-58. DOI: 10.1016/j.jim.2016.12.006. View