A high-throughput RNAi screen for detection of immune-checkpoint molecules that mediate tumor resistance to cytotoxic T lymphocytes

EMBO Mol Med. 2015 Apr;7(4):450-63. doi: 10.15252/emmm.201404414.

Abstract

The success of T cell-based cancer immunotherapy is limited by tumor's resistance against killing by cytotoxic T lymphocytes (CTLs). Tumor-immune resistance is mediated by cell surface ligands that engage immune-inhibitory receptors on T cells. These ligands represent potent targets for therapeutic inhibition. So far, only few immune-suppressive ligands have been identified. We here describe a rapid high-throughput siRNA-based screening approach that allows a comprehensive identification of ligands on human cancer cells that inhibit CTL-mediated tumor cell killing. We exemplarily demonstrate that CCR9, which is expressed in many cancers, exerts strong immune-regulatory effects on T cell responses in multiple tumors. Unlike PDL1, which inhibits TCR signaling, CCR9 regulates STAT signaling in T cells, resulting in reduced T-helper-1 cytokine secretion and reduced cytotoxic capacity. Moreover, inhibition of CCR9 expression on tumor cells facilitated immunotherapy of human tumors by tumor-specific T cells in vivo. Taken together, this method allows a rapid and comprehensive determination of immune-modulatory genes in human tumors which, as an entity, represent the 'immune modulatome' of cancer.

Keywords: RNAi screen; cancer immunotherapy; immune suppression.

MeSH terms

  • Animals
  • B7-H1 Antigen / immunology
  • CD8-Positive T-Lymphocytes* / immunology
  • CD8-Positive T-Lymphocytes* / pathology
  • Female
  • Humans
  • Immunity, Cellular*
  • Immunotherapy / methods*
  • MCF-7 Cells
  • Mice
  • Neoplasms, Experimental* / immunology
  • Neoplasms, Experimental* / pathology
  • Neoplasms, Experimental* / therapy
  • RNA Interference*
  • Receptors, CCR / immunology
  • Th1 Cells* / immunology
  • Th1 Cells* / pathology

Substances

  • B7-H1 Antigen
  • CC chemokine receptor 9
  • CD274 protein, human
  • Receptors, CCR