Silencing of SIRPα enhances the antitumor efficacy of CAR-M in solid tumors

Cell Mol Immunol. 2024 Nov;21(11):1335-1349. doi: 10.1038/s41423-024-01220-3. Epub 2024 Oct 8.

Abstract

The potential of macrophage-mediated phagocytosis as a cancer treatment is promising. Blocking the CD47-SIRPα interaction with a CD47-specific antibody significantly enhances macrophage phagocytosis. However, concerns regarding their toxicity to nontumor cells remain substantial. Here, we engineered chimeric antigen receptor macrophages (CAR-Ms) by fusing a humanized single-chain variable fragment with FcγRIIa and integrating short hairpin RNA to silence SIRPα, thereby disrupting the CD47-SIRPα signaling pathway. These modified CAR-shSIRPα-M cells exhibited an M1-like phenotype, superior phagocytic function, substantial cytotoxic effects on HER2-positive tumor cells, and the ability to eliminate patient-derived organoids. In vivo, CAR-M cells significantly inhibited tumor growth and prolonged survival in tumor-bearing mice. Notably, CAR-shSIRPα-M cells enhanced cytotoxic T-cell infiltration into tumors, thereby enhancing the antitumor response in both the humanized immune system mouse model and immunocompetent mice. Mechanistically, SIRPα inhibition activated inflammatory pathways and the cGAS-STING signaling cascade in CAR-M cells, leading to increased production of proinflammatory cytokines, reactive oxygen species, and nitric oxide, thereby enhancing their antitumor effects. These findings underscore the potential of SIRPα inhibition as a novel strategy to increase the antitumor efficacy of CAR-M cells in cancer immunotherapy, particularly against solid tumors.

Keywords: CAR-M; Cancer immunotherapy; Phagocytosis; SIRPα; Solid tumor.

MeSH terms

  • Animals
  • Antigens, Differentiation*
  • CD47 Antigen* / metabolism
  • Cell Line, Tumor
  • Female
  • Gene Silencing
  • Humans
  • Immunotherapy, Adoptive / methods
  • Macrophages / immunology
  • Macrophages / metabolism
  • Mice
  • Neoplasms / immunology
  • Neoplasms / therapy
  • Phagocytosis
  • Receptors, Chimeric Antigen* / genetics
  • Receptors, Chimeric Antigen* / immunology
  • Receptors, Chimeric Antigen* / metabolism
  • Receptors, IgG / genetics
  • Receptors, IgG / metabolism
  • Receptors, Immunologic* / genetics
  • Receptors, Immunologic* / metabolism
  • Signal Transduction

Substances

  • Receptors, Immunologic
  • Receptors, Chimeric Antigen
  • SIRPA protein, human
  • CD47 Antigen
  • Antigens, Differentiation
  • Receptors, IgG