Feasibility of controlling CD38-CAR T cell activity with a Tet-on inducible CAR design

PLoS One. 2018 May 30;13(5):e0197349. doi: 10.1371/journal.pone.0197349. eCollection 2018.

Abstract

Recent clinical advances with chimeric antigen receptor (CAR) T cells have led to the accelerated clinical approval of CD19-CARs to treat acute lymphoblastic leukemia. The CAR T cell therapy is nevertheless associated with toxicities, especially if the CARs are not entirely tumor-specific. Therefore, strategies for controlling the CAR T cell activity are required to improve their safety profile. Here, by using the multiple myeloma (MM)-associated CD38 molecule as target molecule, we tested the feasibility and utility of a doxycycline (DOX) inducible Tet-on CD38-CAR design to control the off-target toxicities of CAR T cells. Using CARs with high affinity to CD38, we demonstrate that this strategy allows the proper induction of CD38-CARs and CAR-mediated T cell cytotoxicity in a DOX-dose dependent manner. Especially when the DOX dose was limited to 10ng/ml, its removal resulted in a relatively rapid decay of CAR- related off-tumor effects within 24 hours, indicating the active controllability of undesired CAR activity. This Tet-on CAR design also allowed us to induce the maximal anti-MM cytotoxic activity of affinity-optimized CD38-CAR T cells, which already display a low toxicity profile, hereby adding a second level of safety to these cells. Collectively, these results indicate the possibility to utilize this DOX inducible CAR-design to actively regulate the CAR-mediated activities of therapeutic T cells. We therefore conclude that the Tet-on system may be more advantageous above suicide-genes to control the potential toxicities of CAR T cells without the need to destroy them permanently.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP-ribosyl Cyclase 1 / immunology*
  • ADP-ribosyl Cyclase 1 / metabolism
  • Bone Marrow / metabolism
  • Cell Death
  • Cells, Cultured
  • Doxycycline / pharmacology*
  • Genetic Vectors
  • Humans
  • Leukocytes, Mononuclear / metabolism
  • Multiple Myeloma / metabolism
  • Receptors, Antigen, T-Cell / genetics*
  • Receptors, Antigen, T-Cell / immunology*
  • Receptors, Antigen, T-Cell / metabolism
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism

Substances

  • Receptors, Antigen, T-Cell
  • ADP-ribosyl Cyclase 1
  • Doxycycline

Grants and funding

Esther Drent and Tuna Mutis received financial support from ‘Fonds Stimulans’, a Dutch non-profit foundation stimulating Multiple Myeloma research, The Netherlands. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.