Synergistic effect of ibrutinib and CD19 CAR-T cells on Raji cells in vivo and in vitro

Cancer Sci. 2020 Nov;111(11):4051-4060. doi: 10.1111/cas.14638. Epub 2020 Sep 15.

Abstract

Ibrutinib might improve the efficacy of anti-CD19 chimeric antigen receptor (CD19 CAR) T-cell therapy in chronic lymphocytic leukemia (CLL). We studied the possibility and mechanism of the synergistic effect of ibrutinib and CAR-T cells in other types of lymphoma. In this study, we selected the CD19 CAR-T cells of a patient with lymphoma who failed in his CD19 CAR-T-cell therapy and a dose of 8 mg/kg/d ibrutinib. Subcutaneous and tail vein tumorigenic mice were established with Raji cells. The differences in the synergistic effect between these 2 models were compared by bioluminescence imaging (BLI) monitoring and flow cytometry (FCM). The expression of the STAT-3 signaling pathway was assessed by western blot analysis. There was no synergistic effect of ibrutinib and CD19 CAR-T cells in vitro. Programmed cell death-ligand 1 (PD-L1) was expressed in 0.23 ± 0.06% of Raji cells. In the subcutaneous tumorigenic model, the luciferase signal was reduced significantly in the group receiving ibrutinib combined with CD19 CAR-T cells. Moreover, the proportion of CD19 CAR-T cells was higher in the polytherapy group than in the CAR-T-cell monotherapy group. However, we did not get an analogous synergistic effect in the tail vein tumorigenic model. STAT-3 signaling pathway expression in the residual tumor cells did not differ between those with and those without ibrutinib, suggesting that the IL-10/STAT-3/PD-L1 pathway was not involved in the synergistic effect. Therefore, some other mechanism might be a target for ibrutinib. Our results provide evidence for the use of ibrutinib in polytherapy for other types of B-cell lymphoma.

Keywords: Bruton tyrosine kinase inhibitor; Raji cell line; chimeric antigen receptor; programmed cell death-ligand 1; tumor microenvironment.

MeSH terms

  • Adenine / analogs & derivatives*
  • Adenine / pharmacology
  • Adult
  • Aged
  • Animals
  • Antigens, CD19 / immunology*
  • Biomarkers, Tumor
  • Cell Line, Tumor
  • Combined Modality Therapy
  • Disease Models, Animal
  • Female
  • Humans
  • Immunophenotyping
  • Immunotherapy, Adoptive* / methods
  • Lymphoma, Large B-Cell, Diffuse / genetics
  • Lymphoma, Large B-Cell, Diffuse / immunology
  • Lymphoma, Large B-Cell, Diffuse / pathology
  • Lymphoma, Large B-Cell, Diffuse / therapy
  • Male
  • Mice
  • Middle Aged
  • Neoplasm Staging
  • Piperidines / pharmacology*
  • Protein Kinase Inhibitors / pharmacology*
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Antigen, T-Cell / metabolism
  • STAT3 Transcription Factor / metabolism
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism*
  • Treatment Outcome
  • Xenograft Model Antitumor Assays

Substances

  • Antigens, CD19
  • Biomarkers, Tumor
  • Piperidines
  • Protein Kinase Inhibitors
  • Receptors, Antigen, T-Cell
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • ibrutinib
  • Adenine