Point-Of-Care CAR T-Cell Production (ARI-0001) Using a Closed Semi-automatic Bioreactor: Experience From an Academic Phase I Clinical Trial
Por:
Castella, M, Caballero-Banos, M, Ortiz-Maldonado, V, Gonzalez-Navarro, EA, Sune, G, Antonana-Vidosola, A, Boronat, A, Marzal, B, Millan, L, Martin-Antonio, B, Cid, J, Lozano, M, Garcia, E, Tabera, J, Trias, E, Perpina, U, Canals, JM, Baumann, T, Benitez-Ribas, D, Campo, E, Yague, J, Urbano-Ispizua, A, Rives, S, Delgado, J, Juan, M
Publicada:
20 mar 2020
Resumen:
Development of semi-automated devices that can reduce the hands-on time and standardize the production of clinical-grade CAR T-cells, such as CliniMACS Prodigy from Miltenyi, is key to facilitate the development of CAR T-cell therapies, especially in academic institutions. However, the feasibility of manufacturing CAR T-cell products from heavily pre-treated patients with this system has not been demonstrated yet. Here we report and characterize the production of 28 CAR T-cell products in the context of a phase I clinical trial for CD19+ B-cell malignancies (NCT03144583). The system includes CD4-CD8 cell selection, lentiviral transduction and T-cell expansion using IL-7/IL-15. Twenty-seven out of 28 CAR T-cell products manufactured met the full list of specifications and were considered valid products. Ex vivo cell expansion lasted an average of 8.5 days and had a mean transduction rate of 30.6 +/- 13.44%. All products obtained presented cytotoxic activity against CD19+ cells and were proficient in the secretion of pro-inflammatory cytokines. Expansion kinetics was slower in patient's cells compared to healthy donor's cells. However, product potency was comparable. CAR T-cell subset phenotype was highly variable among patients and largely determined by the initial product. T-CM and T-EM were the predominant T-cell phenotypes obtained. 38.7% of CAR T-cells obtained presented a T-N or T-CM phenotype, in average, which are the subsets capable of establishing a long-lasting T-cell memory in patients. An in-depth analysis to identify individual factors contributing to the optimal T-cell phenotype revealed that ex vivo cell expansion leads to reduced numbers of T-N, T-SCM, and T-EFF cells, while T-CM cells increase, both due to cell expansion and CAR-expression. Overall, our results show for the first time that clinical-grade production of CAR T-cells for heavily pre-treated patients using CliniMACS Prodigy system is feasible, and that the obtained products meet the current quality standards of the field. Reduced ex vivo expansion may yield CAR T-cell products with increased persistence in vivo.
Filiaciones:
Castella, M:
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hematol, Barcelona, Spain
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Blood & Tissue Bank BST, Barcelona, Spain
Caballero-Banos, M:
Hosp Clin Barcelona, Ctr Diagnost Biomed, Dept Immunol, Barcelona, Spain
Hosp St Joan Deu, Barcelona, Spain
Ortiz-Maldonado, V:
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hematol, Barcelona, Spain
Gonzalez-Navarro, EA:
Hosp Clin Barcelona, Ctr Diagnost Biomed, Dept Immunol, Barcelona, Spain
Sune, G:
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hematol, Barcelona, Spain
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Antonana-Vidosola, A:
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hematol, Barcelona, Spain
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Boronat, A:
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Hosp Clin Barcelona, Ctr Diagnost Biomed, Dept Immunol, Barcelona, Spain
Marzal, B:
Hosp Clin Barcelona, Ctr Diagnost Biomed, Dept Immunol, Barcelona, Spain
Millan, L:
Hosp Clin Barcelona, Ctr Diagnost Biomed, Dept Immunol, Barcelona, Spain
Martin-Antonio, B:
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hematol, Barcelona, Spain
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Cid, J:
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hemotherapy & Hemostasis, Barcelona, Spain
Lozano, M:
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hemotherapy & Hemostasis, Barcelona, Spain
Garcia, E:
Blood & Tissue Bank BST, Barcelona, Spain
Hosp St Joan Deu Barcelona, Apheresis Unit, Barcelona, Spain
Tabera, J:
Blood & Tissue Bank BST, Barcelona, Spain
Hosp Clin Barcelona, Unit Adv Therapies, Barcelona, Spain
Trias, E:
Hosp Clin Barcelona, Unit Adv Therapies, Barcelona, Spain
Perpina, U:
Univ Barcelona, Prod & Validat Ctr Adv Therapies Creatio, Dept Biomed Sci, Stem Cells & Regenerat Med Lab, Barcelona, Spain
Canals, JM:
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Univ Barcelona, Prod & Validat Ctr Adv Therapies Creatio, Dept Biomed Sci, Stem Cells & Regenerat Med Lab, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Baumann, T:
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hematol, Barcelona, Spain
Benitez-Ribas, D:
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Hosp Clin Barcelona, Ctr Diagnost Biomed, Dept Immunol, Barcelona, Spain
Campo, E:
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Hosp Clin Barcelona, Dept Pathol, IDIBAPS, Barcelona, Spain
Ctr Invest Biomed Red Canc, Barcelona, Spain
Inst Catalana Recerca & Estudis Avancats, Barcelona, Spain
Yague, J:
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Hosp Clin Barcelona, Ctr Diagnost Biomed, Dept Immunol, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Urbano-Ispizua, A:
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hematol, Barcelona, Spain
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Univ Barcelona, Sch Med, Josep Carreras Leukemia Res Inst, Dept Biomed, Barcelona, Spain
Hosp Clin Barcelona, Immunotherapy Unit, Blood & Tissue Bank, Barcelona, Spain
Rives, S:
Hosp St Joan de Deu, Dept Pediat Hematol & Oncol, Barcelona, Spain
Inst Recerca St Joan de Deu, Barcelona, Spain
Delgado, J:
Hosp Clin Barcelona, Inst Clin Malalties Hematol & Oncol, Dept Hematol, Barcelona, Spain
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Ctr Invest Biomed Red Canc, Barcelona, Spain
Juan, M:
Inst Invest Biomed August Pi & Sunyer IDIBAPS, Barcelona, Spain
Blood & Tissue Bank BST, Barcelona, Spain
Hosp Clin Barcelona, Ctr Diagnost Biomed, Dept Immunol, Barcelona, Spain
Hosp St Joan Deu, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Hosp Clin Barcelona, Immunotherapy Unit, Blood & Tissue Bank, Barcelona, Spain
Green Published, gold
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