Rodriguez-Marquez, Calleja-Cervantes, and Serrano et al. addressed the impact of CAR density on BCMA CAR T cell antitumor activity and regulation. Functional assays and scRNAseq revealed that a high CAR density on T cells (CARHighT) resulted in increased signaling in the absence of antigen (tonic signaling), increased activation and upregulation of exhaustion gene markers, and decreased bone marrow infiltration in vivo, compared to CARLowT cells. A CARHighT gene signature was shown to be associated with a worse clinical response to CAR T cell therapy in several hematological malignancies, suggesting translational relevance in patients treated with CAR T cells.
Contributed by Katherine Turner
ABSTRACT: Identification of new markers associated with long-term efficacy in patients treated with CAR T cells is a current medical need, particularly in diseases such as multiple myeloma. In this study, we address the impact of CAR density on the functionality of BCMA CAR T cells. Functional and transcriptional studies demonstrate that CAR T cells with high expression of the CAR construct show an increased tonic signaling with up-regulation of exhaustion markers and increased in vitro cytotoxicity but a decrease in in vivo BM infiltration. Characterization of gene regulatory networks using scRNA-seq identified regulons associated to activation and exhaustion up-regulated in CAR(High) T cells, providing mechanistic insights behind differential functionality of these cells. Last, we demonstrate that patients treated with CAR T cell products enriched in CAR(High) T cells show a significantly worse clinical response in several hematological malignancies. In summary, our work demonstrates that CAR density plays an important role in CAR T activity with notable impact on clinical response.
Author Info: (1) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (2) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Computational
Author Info: (1) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (2) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Computational Biology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (3) Computational Biology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (4) Department of Hematology, Hospital Clinic de Barcelona, IDIBAPS, Universidad de Barcelona, Barcelona, Spain. (5) Hematology and Cell Therapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. (6) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (7) Hematology and Cell Therapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. (8) Department of Immunology, Hospital Clinic de Barcelona, IDIBAPS, Universidad de Barcelona, Barcelona, Spain. (9) Hematology Service, Hospital Universitario de Navarra, IdiSNA, Pamplona, Spain. (10) Immunology and Immunotherapy Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (11) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (12) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. (13) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (14) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. (15) Hematology and Cell Therapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. (16) Flow Cytometry Core, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (17) Hematology Service, Hospital Universitario de Navarra, IdiSNA, Pamplona, Spain. (18) Hematology Service, Hospital Universitario de Navarra, IdiSNA, Pamplona, Spain. (19) Department of Immunology, Hospital Clinic de Barcelona, IDIBAPS, Universidad de Barcelona, Barcelona, Spain. (20) Department of Hematology, Hospital Clinic de Barcelona, IDIBAPS, Universidad de Barcelona, Barcelona, Spain. (21) Department of Immunology, Hospital Clinic de Barcelona, IDIBAPS, Universidad de Barcelona, Barcelona, Spain. Immunotherapy platform Hospital Sant Joan de Déu, Barcelona, Spain. (22) Department of Hematology, Hospital Clinic de Barcelona, IDIBAPS, Universidad de Barcelona, Barcelona, Spain. (23) Hematology and Cell Therapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. (24) Hematology and Cell Therapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. (25) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. Flow Cytometry Core, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (26) Immunology and Immunotherapy Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. (27) Hematology and Cell Therapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. Immunology and Immunotherapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. (28) Hematology and Cell Therapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. Immunology and Immunotherapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. (29) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Hematology and Cell Therapy Department, Clnica Universidad de Navarra (CUN), Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. Cancer Center Universidad de Navarra (CCUN), Pamplona, Spain. (30) Department of Hematology, Hospital Clinic de Barcelona, IDIBAPS, Universidad de Barcelona, Barcelona, Spain. (31) Computational Biology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. Data Science and Artificial Intelligence Institute (DATAI), Universidad de Navarra, Pamplona, Spain. (32) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. (33) Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, Pamplona, Spain. Hematology and Cell Therapy Department, Clínica Universidad de Navarra (CUN), Pamplona, Spain. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain. Cancer Center Universidad de Navarra (CCUN), Pamplona, Spain.
