Logic-Gated ROR1 Chimeric Antigen Receptor Expression Rescues T Cell-Mediated Toxicity to Normal Tissues and Enables Selective Tumor Targeting

Cancer Cell. 2019 Mar 18;35(3):489-503.e8. doi: 10.1016/j.ccell.2019.02.003.


Many potential targets for CAR-T cells in solid tumors are expressed in some normal tissues, raising concern for off-tumor toxicity. Following lymphodepletion, CAR-T cells targeting the tumor-associated antigen ROR1 lysed tumors in mice but induced lethal bone marrow failure due to recognition of ROR1+ stromal cells. To improve selectivity, we engineered T cells with synthetic Notch (synNotch) receptors specific for EpCAM or B7-H3, which are expressed on ROR1+ tumor cells but not ROR1+ stromal cells. SynNotch receptors induced ROR1 CAR expression selectively within the tumor, resulting in tumor regression without toxicity when tumor cells were segregated from, but not when co-localized with, normal ROR1+ cells. This strategy, thus, permits safe targeting of tumors that are sufficiently separated from normal cells.

Keywords: B7-H3; ROR1; T cells; chimeric antigen receptor; combinatorial antigen recognition; immunotherapy; logic gating; mesenchymal stem cell; synthetic Notch receptors; toxicity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Immunotherapy, Adoptive / methods*
  • K562 Cells
  • Mice
  • Neoplasms / immunology
  • Neoplasms / therapy*
  • Receptor Tyrosine Kinase-like Orphan Receptors / genetics*
  • Receptor Tyrosine Kinase-like Orphan Receptors / metabolism
  • Receptors, Chimeric Antigen / immunology*
  • T-Lymphocytes / immunology
  • Xenograft Model Antitumor Assays


  • Receptors, Chimeric Antigen
  • ROR1 protein, human
  • Receptor Tyrosine Kinase-like Orphan Receptors