A Chemical Switch System to Modulate Chimeric Antigen Receptor T Cell Activity through Proteolysis-Targeting Chimaera Technology

ACS Synth Biol. 2020 May 15;9(5):987-992. doi: 10.1021/acssynbio.9b00476. Epub 2020 Apr 30.

Abstract

Despite the excellent efficacy of chimeric antigen receptor (CAR T) cell therapy, concerns about its safety have been constantly raised. The side effects of CAR T cells result from an aberrantly upregulation of CAR T cell activity. Therefore, it is crucial to control the CAR T cell activity whenever the patient is at risk. For this purpose, the iCas9 system, which induces apoptosis in CAR T cell through caspase-9 dimerization by compound, has been invented and is currently going under clinical trial. However, the iCas9 system is irreversible, as the entire CAR T cell population is removed from the patient. Thus, CAR T cells, which are very expensive, should be reinfused to the patients after they recovered from the side-effect. Here, we propose a new CAR T cell safety strategy, which targets CAR "protein", not CAR "T cell". In this system, the CAR construct is modified to bear a bromodomain (BD). The addition of a BD in the CAR protein did not interfere with the original CAR functions, such as cytokine secretion and target cell lysis. Our data showed that the use of a proteolysis-targeting chimaera (PROTAC) compound against BD successfully degraded the BD-containing CAR protein. Moreover, the CAR expression is recovered when the PROTAC compound is removed from the cell, demonstrating that our system is reversible. In a target cell lysis assay, the PROTAC compound successfully suppressed the lytic activity of CAR T cells by degrading the CAR protein. In conclusion, we developed a new safety system in which CAR T cells can be "reversibly" controlled by a compound.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Azepines / chemistry
  • Azepines / pharmacology
  • Cell Line
  • Histone Acetyltransferases / chemistry
  • Humans
  • Immunotherapy, Adoptive / methods*
  • Interleukin-2 / metabolism
  • Protein Domains
  • Proteolysis / drug effects
  • Receptors, Chimeric Antigen / chemistry
  • Receptors, Chimeric Antigen / genetics
  • Receptors, Chimeric Antigen / metabolism*
  • T-Lymphocytes / cytology
  • T-Lymphocytes / metabolism
  • Thalidomide / analogs & derivatives
  • Thalidomide / chemistry
  • Thalidomide / pharmacology
  • Transcription Factors / chemistry
  • Ubiquitin-Protein Ligases / metabolism

Substances

  • ARV-825
  • Azepines
  • BRD2 protein, human
  • Interleukin-2
  • Receptors, Chimeric Antigen
  • Transcription Factors
  • Thalidomide
  • BRD1 protein, human
  • Histone Acetyltransferases
  • Ubiquitin-Protein Ligases