Sobral-da-Costa, E. (Elaine)

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    Automated database-guided expert-supervised orientation for immunophenotypic diagnosis and classification of acute leukemia
    (Springer Nature, 2018) Verde, J. (Javier); Marvelde, J. (Jeroen) te; Grigore, G. (Georgiana); Martin-Ayuso, M. (M.); Asnaf, V. (V.); Novakova, M. (Michaela); Buracchi, C. (Chiara); Fernandez, P. (P.); Bulsa, J. (J.); Orfao, A. (Alberto); Vidriales, M.B. (María Belén); Trinquand, A. (A.); Kalina, T. (Tomas); Mejstrikova, E. (Ester); Sedek, L. (Lukasz); Bras, A.E. (Anne E.); Matarraz, S. (Sergio); Lhermitte, L. (Ludovic); Sobral-da-Costa, E. (Elaine); Szczepanski, T. (Tomasz); Hrusak, O. (O.); Burgos, L. (Leire); Brüggemann, M. (Monika); López, A. (Andrés); Dongen, J.J.M. (Jacques J. M.) van; Paiva, B. (Bruno); Gaipa, G. (Giuseppe); Sonneveld, E. (E.); Sluijs-Gelling, A. (Alita) van der; Lecrevisse, Q. (Quentin); Sá-Bacelar, T. (T.) de; Velden, V.H.J. (Vicent H. J.) van der; Pedreira, C.E. (Carlos E.)
    Precise classification of acute leukemia (AL) is crucial for adequate treatment. EuroFlow has previously designed an AL orientation tube (ALOT) to guide towards the relevant classification panel (T-cell acute lymphoblastic leukemia (T-ALL), B-cell precursor (BCP)-ALL and/or acute myeloid leukemia (AML)) and final diagnosis. Now we built a reference database with 656 typical AL samples (145 T-ALL, 377 BCP-ALL, 134 AML), processed and analyzed via standardized protocols. Using principal component analysis (PCA)-based plots and automated classification algorithms for direct comparison of single-cells from individual patients against the database, another 783 cases were subsequently evaluated. Depending on the database-guided results, patients were categorized as: (i) typical T, B or Myeloid without or; (ii) with a transitional component to another lineage; (iii) atypical; or (iv) mixed-lineage. Using this automated algorithm, in 781/783 cases (99.7%) the right panel was selected, and data comparable to the final WHO-diagnosis was already provided in >93% of cases (85% T-ALL, 97% BCP-ALL, 95% AML and 87% mixed-phenotype AL patients), even without data on the full-characterization panels. Our results show that database-guided analysis facilitates standardized interpretation of ALOT results and allows accurate selection of the relevant classification panels, hence providing a solid basis for designing future WHO AL classifications.
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    Automated identification of leukocyte subsets improves standardization of database-guided expert-supervised diagnostic orientation in acute leukemia: a EuroFlow study
    (2021) Fluxá, R. (Rafael); Montero, J. (Juan); Grigore, G. (Georgiana); Buracchi, C. (Chiara); Fernández, P. (Paula); Morf, D. (Daniela); Orfao, A. (Alberto); Nierkens, S. (Stefan); Mejstrikova, E. (Ester); Barrena, S. (Susana); Sedek, L. (Lukasz); Bie, M. (Maaike) de; Lhermitte, L. (Ludovic); Sobral-da-Costa, E. (Elaine); Szczepanski, T. (Tomasz); Barreau, S. (Sylvain); Aanei, C.M. (Carmen Mariana); Burgos, L. (Leire); Brüggemann, M. (Monika); Dongen, J.J.M. (Jacques J. M.) van; Caetano, J. (Joana); Gaipa, G. (Giuseppe); Hernández-Delgado, A. (Alejandro); Sluijs-Gelling, A. (Alita) van der; Lecrevisse, Q. (Quentin); Velden, V.H.J. (Vicent H. J.) van der; Pedreira, C.E. (Carlos E.)
    Precise classification of acute leukemia (AL) is crucial for adequate treatment. EuroFlow has previously designed an AL orientation tube (ALOT) to guide toward the relevant classification panel and final diagnosis. In this study, we designed and validated an algorithm for automated (database-supported) gating and identification (AGI tool) of cell subsets within samples stained with ALOT. A reference database of normal peripheral blood (PB,n = 41) and bone marrow (BM;n = 45) samples analyzed with the ALOT was constructed, and served as a reference for the AGI tool to automatically identify normal cells. Populations not unequivocally identified as normal cells were labeled as checks and were classified by an expert. Additional normal BM (n = 25) and PB (n = 43) and leukemic samples (n = 109), analyzed in parallel by experts and the AGI tool, were used to evaluate the AGI tool. Analysis of normal PB and BM samples showed low percentages of checks (<3% in PB, <10% in BM), with variations between different laboratories. Manual analysis and AGI analysis of normal and leukemic samples showed high levels of correlation between cell numbers (r(2) > 0.95 for all cell types in PB andr(2) > 0.75 in BM) and resulted in highly concordant classification of leukemic cells by our previously published automated database-guided expert-supervised orientation tool for immunophenotypic diagnosis and classification of acute leukemia (Compass tool).
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    B-cell regeneration profile and minimal residual disease status in bone marrow of treated multiple myeloma patients
    (2021) Aguilera-Sanz, C. (Carmen); Durie, B. (B.); Montero, J. (Juan); Marvelde, J. (Jeroen) te; Aguilar, C. (Carlos); Pérez-Morán, J. (José); García-Mateo, A. (A.); Mendonca-de-Pontes, R. (Robéria); Corral-Mateos, A. (A.); Orfao, A. (Alberto); Maiolino, A. (Angelo); Mateos, M.V. (María Victoria); Barez, A. (A.); Leoz, P. (Pilar); Sobral-da-Costa, E. (Elaine); Puig, N. (Noemí); Labrador, J. (Jorge); Burgos, L. (Leire); García-Sanchez, O. (O.); Dongen, J.J.M. (Jacques J. M.) van; Paiva, B. (Bruno); EuroFlow-Consortium; Sanoja-Flores, L. (L.); Salgado, A.B. (Anna Beatriz); Pessoa-de-Magalhaes, R.J. (Roberto J.); Velden, V.H.J. (Vicent H. J.) van der
    Simple Summary B-cell regeneration during therapy has been associated with the outcome of multiple myeloma (MM) patients. However, the effects of therapy and hemodilution in bone marrow (BM) B-cell recovery have not been systematically evaluated. Here, we show that hemodilution is present in a significant fraction of MM BM samples, leading to lower total B-cell, B-cell precursor (BCP), and normal plasma cell (nPC) counts. Among MM BM samples, decreased percentages (vs. healthy donors) of BCP, transitional/naive B-cell (TBC/NBC) and nPC populations were observed at diagnosis. BM BCP, but not TBC/NBC, increased after induction therapy. At day+100 post-autolo-gous stem cell transplantation, a greater increase in BCP with recovered TBC/NBC numbers but persistently low memory B-cell and nPC counts were found. At the end of therapy, complete response (CR) BM samples showed higher CD19(-) nPC counts vs. non-CR specimens with no clear association between BM B-cell regeneration profiles and patient outcomes. B-cell regeneration during therapy has been considered as a strong prognostic factor in multiple myeloma (MM). However, the effects of therapy and hemodilution in bone marrow (BM) B-cell recovery have not been systematically evaluated during follow-up. MM (n = 177) and adult (>= 50y) healthy donor (HD; n = 14) BM samples were studied by next-generation flow (NGF) to simultaneously assess measurable residual disease (MRD) and residual normal B-cell populations. BM hemodilution was detected in 41 out of 177 (23%) patient samples, leading to lower total B-cell, B-cell precursor (BCP) and normal plasma cell (nPC) counts. Among MM BM, decreased percentages (vs. HD) of BCP, transitional/naive B-cell (TBC/NBC) and nPC populations were observed at diagnosis. BM BCP increased after induction therapy, whereas TBC/NBC counts remained abnormally low. At day+100 postautologous stem cell transplantation, a greater increase in BCP with recovered TBC/NBC cell numbers but persistently low memory B-cell and nPC counts were found. At the end of therapy, complete response (CR) BM samples showed higher CD19(-) nPC counts vs. non-CR specimens. MRD positivity was associated with higher BCP and nPC percentages. Hemodilution showed a negative impact on BM B-cell distribution. Different BM B-cell regeneration profiles are present in MM at diagnosis and after therapy with no significant association with patient outcome.