Timely Leukapheresis May Interfere with the "Fitness" of Lymphocytes Collected for CAR-T Treatment in High Risk DLBCL Patients

Mirko Farina, Marco Chiarini, Camillo Almici, Eugenia Accorsi Buttini, Francesco Zuccalà, Simone Piva, Irene Volonghi, Loris Poli, Simona Bernardi, Federica Colnaghi, Federica Re, Alessandro Leoni, Nicola Polverelli, Alessandro Turra, Enrico Morello, Anna Galvagni, Daniele Moratto, Duilio Brugnoni, Chiara Cattaneo, Emilio Ferrari, Andrea Bianchetti, Michele Malagola, Alessandro Re, Domenico Russo, Mirko Farina, Marco Chiarini, Camillo Almici, Eugenia Accorsi Buttini, Francesco Zuccalà, Simone Piva, Irene Volonghi, Loris Poli, Simona Bernardi, Federica Colnaghi, Federica Re, Alessandro Leoni, Nicola Polverelli, Alessandro Turra, Enrico Morello, Anna Galvagni, Daniele Moratto, Duilio Brugnoni, Chiara Cattaneo, Emilio Ferrari, Andrea Bianchetti, Michele Malagola, Alessandro Re, Domenico Russo

Abstract

The development of chimeric antigen receptor (CAR)-T cell therapy has revolutionized the treatment of hematological diseases. However, approximately 60% of patients relapse after CAR-T cell therapy, and no clear cause for this failure has been identified. The objective of the Bio-CAR-T BS study (ClinicalTrials.gov: NCT05366569) is to improve our understanding of the lymphocyte harvest to maximize the quality of the CAR-T cell product. Of the 14 patients enrolled, 11 were diagnosed with DLBCL, 2 with PMBCL, and 1 with ALL. Five of 11 DLBCL patients met the criteria for "pre-emptive" Lymphocytes-apheresis (being at high risk of second relapse), and 6 were included in the standard-of-care Lymphocytes-apheresis group. Previous autologous stem cell transplantation (ASCT) and age were significantly different between the two groups. At the time of Lymphocyte-apheresis, patients in the "pre-emptive" group had more "fit" lymphocytes (higher CD4+/CD8+ ratio; higher naïve T cells levels) compared with standard group, probably due to the impact of ASCT. At the same time, also being older than 60 years results in a more "exhausted" lymphocyte profile. Overall, "pre-emptive" Ly-apheresis in DLBCL patients at high risk of relapse appears to be feasible and may allow the timely collection of "fit" lymphocytes for CAR-T cell manufacturing.

Keywords: CAR-T; Lymphocytes fitness; T-cells repertoire; pre-emptive lymphocytoapheresis.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
BIO-CAR-T BS study flow chart. ALL, acute lymphoblastic leukemia; CAR, chimeric antigen receptor; CR, complete remission; DLBCL, diffuse large B-cell lymphoma; PMBCL, primary mediastinal B-cell lymphoma; PR, partial remission.
Figure 2
Figure 2
Ly-subsets in DLBCL patients who received “pre-emptive” (blue) or standard Ly-apheresis (red). (A) CD4+/CD8+ ratio. (B) CD4+ T cell subpopulations evaluated as absolute counts (cells/μL) and as a percentage. (C) CD8+ T cell subpopulations evaluated as absolute counts (cells/μL) and as a percentage. * = statistical significance (i.e., p < 0.05).
Figure 3
Figure 3
Ly-subsets in patients aged ≤60 years old (blue) and >60 years old (red). (A) CD4+/CD8+ ratio. (B) CD4+ T cell subpopulations evaluated as absolute counts (cells/μL) and as a percentage. (C) CD8+ T cell subpopulations evaluated as absolute counts (cells/μL) and as a percentage. * = statistical significance (i.e., p < 0.05); ** = statistical significance with p-value between 0.01 and 0.001. *** = statistical significance with p-value between 0.001 and 0.0001.

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