title: Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma.
abstract: Flow cytometry has become a highly valuable method to monitor minimal residual disease (MRD) and evaluate the depth of complete response (CR) in bone marrow (BM) of multiple myeloma (MM) after therapy. However, current flow-MRD has lower sensitivity than molecular methods and lacks standardization. Here we report on a novel next generation flow (NGF) approach for highly sensitive and standardized MRD detection in MM. An optimized 2-tube 8-color antibody panel was constructed in five cycles of design-evaluation-redesign. In addition, a bulk-lysis procedure was established for acquisition of ⩾107cells/sample, and novel software tools were constructed for automatic plasma cell gating. Multicenter evaluation of 110 follow-up BM from MM patients in very good partial response (VGPR) or CR showed a higher sensitivity for NGF-MRD vs conventional 8-color flow-MRD -MRD-positive rate of 47 vs 34% (P=0.003)-. Thus, 25% of patients classified as MRD-negative by conventional 8-color flow were MRD-positive by NGF, translating into a significantly longer progression-free survival for MRD-negative vs MRD-positive CR patients by NGF (75% progression-free survival not reached vs 7 months; P=0.02). This study establishes EuroFlow-based NGF as a highly sensitive, fully standardized approach for MRD detection in MM which overcomes the major limitations of conventional flow-MRD methods and is ready for implementation in routine diagnostics.
authors.affiliation: Cancer Research Center (IBMCC-CSIC/USAL-IBSAL); Cytometry Service (NUCLEUS) and Department of Medicine, University of Salamanca, Salamanca, Spain (USAL).
authors.forename: J
authors.initials: J
authors.lastname: Flores-Montero
authors.affiliation: Cancer Research Center (IBMCC-CSIC/USAL-IBSAL); Cytometry Service (NUCLEUS) and Department of Medicine, University of Salamanca, Salamanca, Spain (USAL).
authors.forename: L
authors.initials: L
authors.lastname: Sanoja-Flores
authors.affiliation: Clinica Universidad de Navarra; Applied Medical Research Center (CIMA), IDISNA, Pamplona, Spain (UNAV).
authors.forename: B
authors.initials: B
authors.lastname: Paiva
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: N
authors.initials: N
authors.lastname: Puig
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: O
authors.initials: O
authors.lastname: García-Sánchez
authors.affiliation: Second Department of Medicine, University Hospital of Schleswig-Holstein, Campus Kiel, Kiel, Germany (UNIKIEL).
authors.forename: S
authors.initials: S
authors.lastname: Böttcher
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands (EMC).
authors.forename: V H J
authors.initials: VHJ
authors.lastname: van der Velden
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: J-J
authors.initials: JJ
authors.lastname: Pérez-Morán
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: M-B
authors.initials: MB
authors.lastname: Vidriales
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: R
authors.initials: R
authors.lastname: García-Sanz
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: C
authors.initials: C
authors.lastname: Jimenez
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: M
authors.initials: M
authors.lastname: González
authors.affiliation: Department of Hematology, Hospital 12 de Octubre, Madrid, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Martínez-López
authors.affiliation: Cancer Research Center (IBMCC-CSIC/USAL-IBSAL); Cytometry Service (NUCLEUS) and Department of Medicine, University of Salamanca, Salamanca, Spain (USAL).
authors.forename: A
authors.initials: A
authors.lastname: Corral-Mateos
authors.affiliation: Cytognos SL, Salamanca, Spain.
authors.forename: G-E
authors.initials: GE
authors.lastname: Grigore
authors.affiliation: Cytognos SL, Salamanca, Spain.
authors.forename: R
authors.initials: R
authors.lastname: Fluxá
authors.affiliation: Faculty of Medicine, Federal University of Rio de Janeiro and Institute of Pediatrics and Childhood Care, Rio de Janeiro, Brazil.
authors.forename: R
authors.initials: R
authors.lastname: Pontes
authors.affiliation: Department of Hematology, Portuguese Institute of Oncology, Lisbon, Portugal (IPOLFG).
authors.forename: J
authors.initials: J
authors.lastname: Caetano
authors.affiliation: Department of Pediatric Hematology and Oncology, Medical University of Silesia in Katowice, Zabrze, Poland (SUM).
authors.forename: L
authors.initials: L
authors.lastname: Sedek
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: M-C
authors.initials: MC
authors.lastname: Del Cañizo
authors.affiliation: Department of Hematology, Hospital Clinic I Provincial, Barcelona, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Bladé
authors.affiliation: Department of Hematology, Hospital 12 de Octubre, Madrid, Spain.
authors.forename: J-J
authors.initials: JJ
authors.lastname: Lahuerta
authors.affiliation: Department of Hematology, Hospital General de Santa Bárbara, Soria, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Aguilar
authors.affiliation: Department of Hematology, Complejo Asistencial de Ávila, Ávila, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Bárez
authors.affiliation: Department of Hematology, Complejo Asistencial de Segovia, Segovia, Spain.
authors.forename: A
authors.initials: A
authors.lastname: García-Mateo
authors.affiliation: Hematology Department, Hospital Universitario de Burgos, Burgos, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Labrador
authors.affiliation: Cancer Research Center (IBMCC-CSIC/USAL-IBSAL); Cytometry Service (NUCLEUS) and Department of Medicine, University of Salamanca, Salamanca, Spain (USAL).
authors.forename: P
authors.initials: P
authors.lastname: Leoz
authors.affiliation: Department of Hematology, Hospital El Bierzo, Ponferrada, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Aguilera-Sanz
authors.affiliation: Clinica Universidad de Navarra; Applied Medical Research Center (CIMA), IDISNA, Pamplona, Spain (UNAV).
authors.forename: J
authors.initials: J
authors.lastname: San-Miguel
authors.affiliation: Department of Hematology, University Hospital of Salamanca, IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain (HUSAL).
authors.forename: M-V
authors.initials: MV
authors.lastname: Mateos
authors.affiliation: Cedars-Sinai Samuel Oschin Cancer Center, Los Angeles, CA, USA.
authors.forename: B
authors.initials: B
authors.lastname: Durie
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands (EMC).|Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: J J M
authors.initials: JJM
authors.lastname: van Dongen
authors.affiliation: Cancer Research Center (IBMCC-CSIC/USAL-IBSAL); Cytometry Service (NUCLEUS) and Department of Medicine, University of Salamanca, Salamanca, Spain (USAL).
authors.forename: A
authors.initials: A
authors.lastname: Orfao
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000918:Antibody Specificity; D002452:Cell Count; D004867:Equipment Design; D005260:Female; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping / Q000295:instrumentation / Q000379:methods*; D008297:Male; D008875:Middle Aged; D009101:Multiple Myeloma / Q000175:diagnosis* / Q000188:drug therapy / Q000473:pathology; D018365:Neoplasm, Residual; D010950:Plasma Cells / Q000473:pathology*; D012680:Sensitivity and Specificity; D012984:Software; D013048:Specimen Handling; D016896:Treatment Outcome