In vitro functionality and endurance of GMP-compliant point-of-care BCMA.CAR-T cells at different timepoints of cryopreservation

• Verfasst von: Jiang, Genqiao [VerfasserIn]
• Verfasst von: Neuber, Brigitte [VerfasserIn]
• Verfasst von: Hückelhoven-Krauss, Angela [VerfasserIn]
• Verfasst von: Höpken, Uta [VerfasserIn]
• Verfasst von: Ding, Yuntian [VerfasserIn]
• Verfasst von: Sedloev, David [VerfasserIn]
• Verfasst von: Wang, Lei [VerfasserIn]
• Verfasst von: Reichman, Avinoam [VerfasserIn]
• Verfasst von: Eberhardt, Franziska [VerfasserIn]
• Verfasst von: Wermke, Martin [VerfasserIn]
• Verfasst von: Rehm, Armin [VerfasserIn]
• Verfasst von: Müller-Tidow, Carsten [VerfasserIn]
• Verfasst von: Schmitt, Anita [VerfasserIn]
• Verfasst von: Schmitt, Michael [VerfasserIn]
• Titel: In vitro functionality and endurance of GMP-compliant point-of-care BCMA.CAR-T cells at different timepoints of cryopreservation
• Verf.angabe: Genqiao Jiang, Brigitte Neuber, Angela Hückelhoven-Krauss, Uta E. Höpken, Yuntian Ding, David Sedloev, Lei Wang, Avinoam Reichman, Franziska Eberhardt, Martin Wermke, Armin Rehm, Carsten Müller-Tidow, Anita Schmitt and Michael Schmitt
• E-Jahr: 2024
• Jahr: 23 January 2024
• Umfang: 15 S.
• Illustrationen: Illustrationen
• Fussnoten: Gesehen am 11.06.2024
• Titel Quelle: Enthalten in: International journal of molecular sciences
• Ort Quelle: Basel : Molecular Diversity Preservation International, 2000
• Jahr Quelle: 2024
• Band/Heft Quelle: 25(2024), 3, Artikel-ID 1394, Seite 1-15
• ISSN Quelle: 1422-0067
• ISSN Quelle: 1661-6596
• DOI: doi:10.3390/ijms25031394
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: B-cell maturation antigen (BCMA)
• Sach-SW: CAR-T cells
• Sach-SW: multiple myeloma
• Sach-SW: stability and function of CAR-T cells
• Sach-SW: timepoint
• K10plus-PPN: 1891059165

Abstract

The search for target antigens for CAR-T cell therapy against multiple myeloma defined the B-cell maturation antigen (BCMA) as an interesting candidate. Several studies with BCMA-directed CAR-T cell therapy showed promising results. Second-generation point-of-care BCMA.CAR-T cells were manufactured to be of a GMP (good manufacturing practice) standard using the CliniMACS Prodigy® device. Cytokine release in BCMA.CAR-T cells after stimulation with BCMA positive versus negative myeloma cell lines, U266/HL60, was assessed via intracellular staining and flow cytometry. The short-term cytotoxic potency of CAR-T cells was evaluated by chromium-51 release, while the long-term potency used co-culture (3 days/round) at effector/target cell ratios of 1:1 and 1:4. To evaluate the activation and exhaustion of CAR-T cells, exhaustion markers were assessed via flow cytometry. Stability was tested through a comparison of these evaluations at different timepoints: d0 as well as d + 14, d + 90 and d + 365 of cryopreservation. As results, (1) Killing efficiency of U266 cells correlated with the dose of CAR-T cells in a classical 4 h chromium-release assay. There was no significant difference after cryopreservation on different timepoints. (2) In terms of endurance of BCMA.CAR-T cell function, BCMA.CAR-T cells kept their ability to kill all tumor cells over six rounds of co-culture. (3) BCMA.CAR-T cells released high amounts of cytokines upon stimulation with tumor cells. There was no significant difference in cytokine release after cryopreservation. According to the results, BCMA.CAR-T cells manufactured under GMP conditions exerted robust and specific killing of target tumor cells with a high release of cytokines. Even after 1 year of cryopreservation, cytotoxic functions were maintained at the same level. This gives clinicians sufficient time to adjust the timepoint of BCMA.CAR-T cell application to the patient’s course of the underlying disease.