STAT3 Silencing and TLR7/8 Pathway Activation Repolarize and Suppress Myeloid-Derived Suppressor Cells From Breast Cancer Patients

Front Immunol. 2021 Feb 19:11:613215. doi: 10.3389/fimmu.2020.613215. eCollection 2020.

Abstract

Cancer cells escape immune destruction. From this perspective, myeloid-derived suppressor cells (MDSCs), which are immunosuppressive in various cancers including breast cancer (BC), are significant. However, the precise mechanisms are unknown. We isolated HLA-DR-CD33+ MDSCs and CD3+ T cells from BC patients' peripheral blood and healthy donors through MACS and immunophenotyped by flow cytometry. Transfection of short-interfering RNAs and treatment with a TLR7/8 agonist altered pathway activities in vitro. Gene expression was analyzed using qRT-PCR, western blotting, and immunohistochemistry. Our findings showed an association between the progression of BC and increased levels of circulating HLA-DR-CD33+ MDSCs. These cells strongly suppress both autologous and analogous CD3+ T cell proliferation and enter the tumor microenvironment. We also identified increased STAT3 signaling and increased IDO and IL-10 expression in BC-derived MDSCs as immunosuppression mechanisms. Further, STAT3 inhibition and TLR7/8 pathway stimulation reduce the immunosuppressive activity of patient-derived MDSCs on T cells by inducing MDSC repolarization and differentiation into mature myeloid cells. This also alters the expression of critical cytokines and transcription factors in CD3+ T cells and, importantly, reduces breast cancer cells' proliferation. Finally, while chemotherapy is able to significantly reduce circulating MDSCs' level in patients with breast cancer, these MDSCs remained highly T cell-suppressive. We identified a novel molecular mechanism of MDSC-mediated immunosuppression. STAT3 inhibition and TLR7/8 pathway stimulation in MDSCs repolarize and suppress MDSCs from breast cancer patients. This offers new opportunities for BC immunotherapy.

Keywords: STAT3; TLR7/8; breast cancer; myeloid-derived suppressor cells; tumor microenvironment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / immunology*
  • CD3 Complex / immunology
  • Cell Differentiation / immunology
  • Cell Proliferation / physiology
  • Cytokines / immunology
  • Female
  • HLA-DR Antigens / immunology
  • Humans
  • Immune Tolerance / immunology
  • Immunophenotyping / methods
  • Middle Aged
  • Myeloid Cells / immunology
  • Myeloid-Derived Suppressor Cells / immunology*
  • STAT3 Transcription Factor / immunology*
  • Signal Transduction / immunology*
  • T-Lymphocytes / immunology
  • Toll-Like Receptor 7 / immunology*
  • Toll-Like Receptor 8 / immunology*
  • Tumor Microenvironment / immunology

Substances

  • CD3 Complex
  • Cytokines
  • HLA-DR Antigens
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • TLR7 protein, human
  • TLR8 protein, human
  • Toll-Like Receptor 7
  • Toll-Like Receptor 8