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Genomics, Gene Therapy and Proteomics

Engineering CD19-specific T lymphocytes with interleukin-15 and a suicide gene to enhance their anti-lymphoma/leukemia effects and safety

Abstract

T lymphocytes expressing a chimeric antigen receptor (CAR) targeting the CD19 antigen (CAR.19) may be of value for the therapy of B-cell malignancies. Because the in vivo survival, expansion and anti-lymphoma activity of CAR.19+ T cells remain suboptimal even when the CAR contains a CD28 costimulatory endodomain, we generated a novel construct that also incorporates the interleukin-15 (IL-15) gene and an inducible caspase-9-based suicide gene (iC9/CAR.19/IL-15). We found that compared with CAR.19+ T cells, iC9/CAR.19/IL-15+ T cells had: (1) greater numeric expansion upon antigen stimulation (10-fold greater expansion in vitro, and 3- to 15-fold greater expansion in vivo) and reduced cell death rate (Annexin-V+/7-AAD+ cells 10±6% for iC9/CAR.19/IL-15+ T cells and 32±19% for CAR.19+ T cells); (2) reduced expression of the programmed death 1 (PD-1) receptor upon antigen stimulation (PD-1+ cells <15% for iC9/CAR.19/IL-15+ T cells versus >40% for CAR.19+ T cells); and (3) improved antitumor effects in vivo (from 4.7- to 5.4-fold reduced tumor growth). In addition, iC9/CAR.19/IL-15+ T cells were efficiently eliminated upon pharmacologic activation of the suicide gene. In summary, this strategy safely increases the anti-lymphoma/leukemia effects of CAR.19-redirected T lymphocytes and may be a useful approach for treatment of patients with B-cell malignancies.

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Acknowledgements

This work was supported in part from Leukemia and Lymphoma Society Specialized Center of Research (SCOR; Grant no. 7018), the NIH Grants PO1CA94237, P50CA126752 and RO1CA131027, Leukemia and Lymphoma Society Translational Research grants, Doris Duke Charitable Foundation/Clinical Scientist development award and CLL Global Research Foundation.

Author contributions. VH, BS, CQ, MZ and JV performed the experiments. AM and BS performed the animal experiments. GD, BS and VH designed the research and analyzed the data. VH, BS and GD wrote the paper. HEH, CMR and MKB critically reviewed the paper. All authors approved the final version of the paper.

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Correspondence to G Dotti.

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Hoyos, V., Savoldo, B., Quintarelli, C. et al. Engineering CD19-specific T lymphocytes with interleukin-15 and a suicide gene to enhance their anti-lymphoma/leukemia effects and safety. Leukemia 24, 1160–1170 (2010). https://doi.org/10.1038/leu.2010.75

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Keywords

  • chimeric antigen receptor
  • IL-15
  • suicide gene

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