Establishment of a 6-, 8- and 10-color multiparameter flow cytometry assay for the detection of minimal residual disease in multiple myeloma patients: challenging diversity in a straightforward approach

Abstract: Multiple Myeloma (MM) is characterized by the accumulation of aberrant plasma cells (aPCs) in the bone marrow. The use of novel agents for anti-MM treatment has enabled deeper responses to therapy and prolonged progression free (PFS) and overall survival. These advances have resulted in the need for highly sensitive detection methods of residual aPCs. Multiparameter flow cytometry (MFC) allows residual aPC detection with high sensitivity, predicting high-risk patient groups and post therapeutic outcomes, but so far is only utilized in clinical trials because standardized version are very expensive and need special equipment (e.g. software). Therefore, the incorporation of affordable, readily available and standardized minimal residual disease (MRD) tools into clinical practice is highly desired. It may allow the improvement of clinical decisions like: 1.) timing of stem cell transplantation, 2.) need for consolidation, 3.) duration of maintenance, 4) therapy modulations and 5.) early relapse detection.
In this study, a 6-, 8- and 10-color panel, using 12 different markers, was thoroughly validated in different MM cell lines (MMCLs), 298 MM patients samples and 14 healthy individuals (HIs) samples, and included fluorescence-minus-one-controls and dilution assays to evaluate the sensitivity of the panels. Data analysis was accomplished using the BeckmanCoulter software Kaluza®. The study was performed with written consent from all patients and HIs and approval of the University of Freiburg ethics committee.
The antigen expression levels in MMCLs using the 3 panels could be confirmed as reported and were similar throughout all panels. Easy to adapt gating strategies were established, using commercially available software to identify aPCs vs normal PCs (nPCs), with a high MRD sensitivity of 10-5, determined via independent dilution assays. MRD negativity, with a prolonged PFS, was observed in 26% of the total patient cohort compared to the MRD positive group. Deeper analyses of MM, precursor stages and leukapheresis samples confirmed this assay as highly valid and revealed new insight into MM course of treatment.
In summary, these results demonstrate a highly validated and affordable straightforward approach for MRD determination in MM, which represents a good alternative to expensive MRD/EuroFlow assays and allows improved routine MM diagnostics at the University Medical Center Freiburg. Our assay does not use special software and is easy adaptable. Furthermore, our study was one of the first that demonstrated reliable results in patients routinely punctured in- and outside clinical trials with a straightforward, convenient, thoroughly validated and resource-efficient assay

Object Details

• Location: Deutsche Nationalbibliothek Frankfurt am Main
• Extent: Online-Ressource
• Language: Englisch
• Notes: Universität Freiburg, Dissertation, 2019

Classification and Topics

• Keyword: Patients ; Flow cytometry ; Multiple myeloma ; Color

Contributors, Places and Time

• Event: Veröffentlichung
• (where): Freiburg
• (who): Universität
• (when): 2019
• Creator: Woerner, Sandra Maria
• Contributor: Engelhardt, Monika ; Zeiser, Robert ; Albert-Ludwigs-Universität Freiburg. Fakultät für Biologie

Further information

• DOI: 10.6094/UNIFR/151251
• URN: urn:nbn:de:bsz:25-freidok-1512517
• Rights: Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Last update: 14.08.2025, 10:52 AM CEST