Journal Article

Point-Of-Care CAR T-Cell Production (ARI-0001) Using a Closed Semi-automatic Bioreactor: Experience From an Academic Phase I Clinical Trial

Baumann, Tycho, Millán, Lucía, Yagüe, Jordi, Antoñana-Vidósola, Asier, Cid, Joan, Caballero-Baños, Miguel, Canals, Josep Ma, Perpiña, Unai, Martín-Antonio, Beatriz, Lozano, Miquel, Castella, Maria, Suñé, Guillermo, Marzal, Berta, Rives, Susana, Urbano-Ispizua, Álvaro, Trias, Esteve, Juan, Manel, Boronat, Anna, Benítez-Ribas, Daniel, García, Enric, González-Navarro, Europa Azucena, Ortiz-Maldonado, Valentín, Tabera, Jaime, Campo, Elías, Delgado, Julio

2020

Overview

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. 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 and T were the predominant T-cell phenotypes obtained. 38.7% of CAR T-cells obtained presented a T or T 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 cell expansion leads to reduced numbers of T , T , and T cells, while T 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 expansion may yield CAR T-cell products with increased persistence .

• Publisher: Frontiers Media SA,Frontiers Media S.A
• Subject: Academic Medical Centers / Adolescent / Adult / Antibodies / Antigens / Apheresis / Automation / Bioreactors / Cancer / CAR T-cell production / CD19 / CD19 antigen / CD4 antigen / CD8 antigen / Cell culture / Cell Proliferation / Cell therapy / Cells, Cultured / Child / chimeric antigen receptor / Clinical trials / Cytotoxicity / Cytotoxicity, Immunologic / Disease / Female / Hematology / Humans / Immunologic Memory / Immunological memory / Immunology / immunotherapy / Immunotherapy, Adoptive - methods / Leukemia / Lymphocytes B / Lymphocytes T / Lymphoma / Male / Malignancy / Medical research / Patients / Pediatrics / Phenotypes / Point-of-Care Systems / Prodigies / Radiation therapy / Response rates / T-Lymphocyte Subsets - immunology / T-Lymphocytes - immunology / Tumors / Young Adult