Optimization of manufacturing conditions for chimeric antigen receptor T cells to favor cells with a central memory phenotype |
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| Institution: | 1. Translational Oncology Laboratory, Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, South Australia, Australia;2. Cancer Clinical Trials Unit, Royal Adelaide Hospital, Adelaide, South Australia, Australia;3. School of Medicine, the University of Adelaide, Adelaide, South Australia, Australia;1. Institute of Child Health, Molecular and Cellular Immunology Unit, University College London, London, UK;2. University College London Cancer Institute, London, UK;3. Research & Development, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany;1. Research Department of Haematology, University College London, London, UK;2. Department of Haematology, University College London Hospitals National Health Service (NHS) Foundation Trust, London |
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| Abstract: | BackgroundChimeric antigen receptor (CAR)-T cells are genetically engineered to recognize tumor-associated antigens and have potent cytolytic activity against tumors. Adoptive therapy with CAR-T cells has been highly successful in B-cell leukemia and lymphoma. However, in solid tumor settings, CAR-T cells face a particularly hostile tumor microenvironment where multiple immune suppressive factors serve to thwart the anti-cancer immune response. Clinical trials of solid tumor antigen-targeted CAR-T cells have shown limited efficacy, and issues for current CAR-T cell therapies include failures of expansion and persistence, tumor entry, deletion and functional exhaustion.MethodsWe compared our standard protocol for CAR-T cell manufacturing, currently used to generate CAR-T cells for a phase 1 clinical trial, with two alternative approaches for T-cell activation and expansion. The resulting cultures were analyzed using multicolor flow cytometry, cytokine bead array and xCELLigence cytotoxicity assays.ResultsWe have found that by changing the method of activation we can promote generation of CAR-T cells with enhanced CD62L and CCR7 expression, increased interleukin (IL)-2 production and retention of cytolytic activity, albeit with slower kinetics.DiscussionWe propose that these phenotypic characteristics are consistent with a central memory phenotype that will better enable CAR-T cell survival and persistence after activation in vivo, and we aim to test this in a continuation of our current phase 1 clinical trial of CAR-T cells in patients with advanced melanoma. |
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