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

Frontiers in Immunology

Jordi Yagüe¹  Josep Ma Canals¹  Julio Delgado³  Elías Campo⁴  Álvaro Urbano-Ispizua⁶  Manel Juan⁶  Susana Rives⁸  Enric García⁹  Maria Castella^1,10  Valentín Ortiz-Maldonado^1,11  Tycho Baumann^1,11  Joan Cid^1,12  Miquel Lozano^1,12  Berta Marzal^1,13  Europa Azucena González-Navarro^1,13  Lucía Millán^1,13  Anna Boronat^1,13  Daniel Benítez-Ribas^1,13  Miguel Caballero-Baños^1,14  Asier Antoñana-Vidósola^1,15  Beatriz Martín-Antonio^1,15  Guillermo Suñé^1,15  Unai Perpiña^1,16  Jaime Tabera^1,17  Esteve Trias^1,17 

[1] 0Universitat de Barcelona, Barcelona, Spain;1Department of Pathology, Hospital Clínic de Barcelona, IDIBAPS, Barcelona, Spain;2Centro de Investigación Biomedical en Red de Cancer, Barcelona, Spain;3Institució Catalana de Recerca i Estudis Avancats, Barcelona, Spain;4Department of Biomedicine, School of Medicine, Josep Carreras Leukemia Research Institute, Universitat de Barcelona, Barcelona, Spain;5Immunotherapy Unit Blood and Tissue Bank-Hospital Clínic de Barcelona, Barcelona, Spain;6Department of Pediatric Hematology and Oncology, Hospital Sant Joan de Déu, Barcelona, Spain;7Institut de Recerca Sant Joan de Déu, Barcelona, Spain;Apheresis Unit, Hospital Sant Joan de Déu de Barcelona, Barcelona, Spain;Blood and Tissue Bank (BST), Barcelona, Spain;Department of Hematology, Institut Clínic de Malalties Hematològiques i Oncològiques, Hospital Clínic de Barcelona, Barcelona, Spain;Department of Hemotherapy and Hemostasis, Institut Clínic de Malalties Hematològiques i Oncològiques, Hospital Clínic de Barcelona, Barcelona, Spain;Department of Immunology, Centro de Diagnóstico Biomédico, Hospital Clínic de Barcelona, Barcelona, Spain;Hospital Sant Joan de Déu, Barcelona, Spain;Institut D'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain;Stem Cells and Regenerative Medicine Laboratory, Department of Biomedical Sciences, Production and Validation Center of Advanced Therapies (Creatio), Universitat de Barcelona, Barcelona, Spain;Unit of Advanced Therapies, Hospital Clínic de Barcelona, Barcelona, Spain;

关键词: chimeric antigen receptor;    CD19;    leukemia;    lymphoma;    immunotherapy;    CAR T-cell production;   

DOI  :  10.3389/fimmu.2020.00482

来源: DOAJ

【 摘 要 】

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. TCM and TEM were the predominant T-cell phenotypes obtained. 38.7% of CAR T-cells obtained presented a TN or TCM 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 TN, TSCM, and TEFF cells, while TCM 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.

【 授权许可】

Unknown