Strong expansion of normal CD19‐negative B‐cell precursors after the use of blinatumomab in the first‐line therapy of acute lymphoblastic leukaemia in children

Mikhailova, Ekaterina; Roumiantseva, Julia; Illarionova, Olga; Lagoyko, Svetlana; Miakova, Natalia; Zerkalenkova, Elena; Zharikova, Liudmila; Olshanskaya, Yulia; Novichkova, Galina; Maschan, Michael; Henze, Guenter; Karachunskiy, Alexander; Popov, Alexander

doi:10.1111/bjh.17760

Cited by 7 publications

(8 citation statements)

References 15 publications

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“…20,28 Taken together with changes in expression of other antigens from MRD-oriented panels, 20,21 specific regeneration patterns 13 and possible lineage switching, 29,30 MFC-MRD after CD19-directed therapy may need to be confirmed with additional tests, especially when blinatumomab is implemented as front-line therapy for BCP-ALL. 26,31 Similarly, the pivotal role of MRD data in the definition of remission achievement, 32 risk group stratification 10 and imminent relapse prediction 32 in primary BCP-ALL treated with chemotherapy alone requires absolute confidence in MFC data. Treatment intensity reduction, which is one of the main trends in contemporary treatment strategies, 33,34 can lead to the appearance of normal BCPs early during therapy, even at the end of induction.…”

Section: Discussionmentioning

confidence: 99%

“…Immunophenotypic changes induced by CD19‐directed agents (CAR‐T or antibody therapy) render MFC‐MRD analysis more sophisticated, as possible loss of CD19 on leukemic cells 20,21,27 significantly changes the gating strategy for proper evaluation of the B‐lineage compartment 20,28 . Taken together with changes in expression of other antigens from MRD‐oriented panels, 20,21 specific regeneration patterns 13 and possible lineage switching, 29,30 MFC‐MRD after CD19‐directed therapy may need to be confirmed with additional tests, especially when blinatumomab is implemented as front‐line therapy for BCP‐ALL 26,31 . Similarly, the pivotal role of MRD data in the definition of remission achievement, 32 risk group stratification 10 and imminent relapse prediction 32 in primary BCP‐ALL treated with chemotherapy alone requires absolute confidence in MFC data.…”

Section: Discussionmentioning

confidence: 99%

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Flow cell sorting followed by PCR‐based clonality testing may assist in questionable diagnosis and monitoring of acute lymphoblastic leukemia

Semchenkova

Zhogov

Захарова

et al. 2023

Int J Lab Hematology

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Introduction Multicolor flow cytometry (MFC) has highly reliable and flexible algorithms for diagnosis and monitoring of acute lymphoblastic leukemia (ALL). However, MFC analysis can be affected by poor sample quality or novel therapeutic options (e.g., targeted therapies and immunotherapy). Therefore, an additional confirmation of MFC data may be needed. We propose a simple approach for validation of MFC findings in ALL by sorting questionable cells and analyzing immunoglobulin/T‐cell receptor (IG/TR) gene rearrangements via EuroClonality‐based multiplex PCR. Patients and Methods We obtained questionable MFC results for 38 biological samples from 37 patients. In total, 42 cell populations were isolated by flow cell sorting for downstream multiplex PCR. Most of the patients (n = 29) had B‐cell precursor ALL and were investigated for measurable residual disease (MRD); 79% of them received CD19‐directed therapy (blinatumomab or CAR‐T). Results We established the clonal nature of 40 cell populations (95.2%). By using this technique, we confirmed very low MRD levels (<0.01% MFC‐MRD). We also applied it to several ambiguous findings for diagnostic samples, including those with mixed‐phenotype acute leukemia, and the results obtained impacted the final diagnosis. Conclusion We have demonstrated possibilities of a combined approach (cell sorting and PCR‐based clonality assessment) to validate MFC findings in ALL. The technique is easy to implement in diagnostic and monitoring workflows, as it does not require isolation of a large number of cells and knowledge of individual clonal rearrangements. We believe it provides important information for further treatment.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Flow cell sorting followed by PCR‐based clonality testing may assist in questionable diagnosis and monitoring of acute lymphoblastic leukemia

Semchenkova

Zhogov

Захарова

et al. 2023

Int J Lab Hematology

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“…The examples of B‐lineage compartment gating using main described routes in follow‐up samples are shown on Figure 4. Moreover, using antigens which appear very early in B‐lineage development, it is necessary also to adjust gating strategy to the possible appearance of normal CD19‐negative progenitors, found to be detectable frequently after blinatumomab and CAR‐T application (Mikhailova, Roumiantseva, et al, 2022; Mikhailova, Semchenkova, et al, 2021). Applicability of the described algorithm for MFC‐MRD monitoring after CD19‐directed therapy with comparison to IG/TR gene rearrangements by NGS and/or fusion gene transcript monitoring by molecular techniques is currently under investigation in our laboratory (manuscript in preparation).…”

Section: Discussionmentioning

confidence: 99%

B‐lineage antigens that are useful to substitute CD19 for minimal residual disease monitoring in B cell precursor acute lymphoblastic leukemia after CD19 targeting

Mikhailova

Itov

Zerkalenkova

et al. 2022

Cytometry Part B Clinical

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Background: The potential loss of CD19 during targeted treatment of B cell precursor acute lymphoblastic leukemia (BCP-ALL) can hamper flow cytometric minimal residual disease (MRD) monitoring. In the current study, we present expression data for antigens that are candidates for CD19 substitution: surface CD22, CD24, CD10, and intracellular (i) CD79a.Methods: Bone marrow samples from 519 consecutive children (below 18 y.o.) with primary BCP-ALL were studied with a focus on expression of CD19, CD10, CD22, CD24, and iCD79a. As these antigens are planned to be used as substitutions for CD19 for primary B cell gating, only total expression on the leukemic population (≥95% cells) was considered appropriate.Results: It was found that each of these antigens is totally expressed in nearly 90% of patients. For each single marker, a subgroup of patients without complete positivity presented with BCP-ALL harboring diverse cytogenetic and molecular genetic aberrations. Based on expression data, we have developed algorithm of simultaneous application of these antigens for initial B-lineage compartment gating, that is applicable for nearly all patients after CD19 targeting. Conclusion:We conclude that the addition of CD22, CD24, and iCD79a to the conventional antibody panel and their application together with CD10 allow for the identification of B-lineage compartment including residual tumor blasts, for MFC-MRD searching in virtually all patients with BCP-ALL after CD19-directed treatment.

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“…Based on the published data from other groups [ 15 , 16 , 17 , 50 ] and our consecutive studies [ 18 , 19 , 20 , 22 , 23 , 24 ], we have developed, validated and implemented an 11-color single-tube approach for MFC-MRD detection in routine practice. Ready-to-use premanufactured tubes containing the majority of used antibodies can seriously improve the reliability of the assay by minimizing possible pipetting errors and providing highly stable cytometric data, which can be analyzed not only manually but also with programmed approaches [ 28 , 51 , 52 ].…”

Section: Discussionmentioning

confidence: 99%

“…We have described the expression of MRD-related antigens by these early BCPs and found their relative expansion in R/R BCP-ALL patients treated with either blinatumomab or CAR-T cells [ 22 ]. Moreover, in children who received blinatumomab during frontline treatment immediately after induction, normal CD19-negative BCPs in substantial quantities were detected in all patients at all evaluated time points [ 23 ]. Thus, the described changes in the normal background inside the B-lineage compartment should also be considered during MFC data analysis and interpretation.…”

Section: Methodsmentioning

confidence: 99%

Reliable Flow-Cytometric Approach for Minimal Residual Disease Monitoring in Patients with B-Cell Precursor Acute Lymphoblastic Leukemia after CD19-Targeted Therapy

Mikhailova

Illarionova

Komkov

et al. 2022

Cancers

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We aimed to develop an antibody panel and data analysis algorithm for multicolor flow cytometry (MFC), which is a reliable method for minimal residual disease (MRD) detection in patients with B-cell precursor acute lymphoblastic leukemia (BCP-ALL) treated with CD19-directed therapy. The development of the approach, which was adapted for the case of possible CD19 loss, was based on the additional B-lineage marker expression data obtained from a study of primary BCP-ALL patients, an analysis of the immunophenotypic changes that occur during blinatumomab or CAR-T therapy, and an analysis of very early CD19-negative normal BCPs. We have developed a single-tube 11-color panel for MFC-MRD detection. CD22- and iCD79a-based primary B-lineage gating (preferably consecutive) was recommended. Based on patterns of antigen expression changes and the relative expansion of normal CD19-negative BCPs, guidelines for MFC data analysis and interpretation were established. The suggested approach was tested in comparison with the molecular techniques: IG/TR gene rearrangement detection by next-generation sequencing (NGS) and RQ-PCR for fusion-gene transcripts (FGTs). Qualitative concordance rates of 82.8% and 89.8% were obtained for NGS-MRD and FGT-MRD results, respectively. We have developed a sensitive and reliable approach that allows MFC-MRD monitoring after CD19-directed treatment, even in the case of possible CD19 loss.

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Strong expansion of normal CD19‐negative B‐cell precursors after the use of blinatumomab in the first‐line therapy of acute lymphoblastic leukaemia in children

Cited by 7 publications

References 15 publications

Flow cell sorting followed by PCR‐based clonality testing may assist in questionable diagnosis and monitoring of acute lymphoblastic leukemia

Flow cell sorting followed by PCR‐based clonality testing may assist in questionable diagnosis and monitoring of acute lymphoblastic leukemia

B‐lineage antigens that are useful to substitute CD19 for minimal residual disease monitoring in B cell precursor acute lymphoblastic leukemia after CD19 targeting

Reliable Flow-Cytometric Approach for Minimal Residual Disease Monitoring in Patients with B-Cell Precursor Acute Lymphoblastic Leukemia after CD19-Targeted Therapy

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