Synthetic SIGLEC9-based chimeric switch receptor augments the efficacy of CAR macrophages against glioblastoma

Proc Natl Acad Sci U S A. 2026 Mar 24;123(12):e2519819123. doi: 10.1073/pnas.2519819123. Epub 2026 Mar 17.

Abstract

Chimeric antigen receptor macrophage (CAR-M) therapy represents a promising therapeutic approach for treating glioblastoma multiforme (GBM). However, durable antitumorigenic macrophage phenotype of CAR-Ms is limited by the highly immunosuppressive tumor microenvironment (TME), wherein Siglec-sialic acid signaling directly drives macrophage polarization toward a protumorigenic phenotype. We here report an in situ synthetic SIGLEC9-based chimeric switch receptor (CSR) for diverting the inhibitory signal into positive ones, augmenting the sustained proinflammatory phenotype and tumoricidal immunity of CAR-Ms in the GBM niche. Specifically, our results showed that macrophage-targeted ionizable lipid nanoparticles efficiently introduce dual circRNAs into macrophages to generate CSR functionalized CAR-Ms in vitro and in vivo. The modified macrophages maintained a proinflammatory state, exhibited superior phagocytic activity, resulting in rapid and efficient eradication of IL13Rα2-positive tumor cells. Moreover, an injectable nanoparticle-hydrogel system for reprogramming macrophages surrounding the glioma resection cavity initiated a locoregional antitumor immune response and elicited robust long-term immunological memory, inhibiting tumor relapse in the postoperative GBM model. In sum, our findings establish that the engineered SIGLEC9-based CSR significantly promotes the maintenance of an antitumoral phenotype of CAR-Ms in the hypersialylated acidic TME, contributing to the improvement of engineered macrophage-based immunotherapy against GBM.

Keywords: SIGLEC9; chimeric antigen receptor macrophages; chimeric switch receptor; glioblastoma.

MeSH terms

  • Animals
  • Antigens, CD
  • Brain Neoplasms* / immunology
  • Brain Neoplasms* / therapy
  • Cell Line, Tumor
  • Glioblastoma* / immunology
  • Glioblastoma* / pathology
  • Glioblastoma* / therapy
  • Humans
  • Immunotherapy, Adoptive* / methods
  • Macrophages* / immunology
  • Macrophages* / metabolism
  • Mice
  • Nanoparticles / chemistry
  • Receptors, Chimeric Antigen* / genetics
  • Receptors, Chimeric Antigen* / immunology
  • Receptors, Chimeric Antigen* / metabolism
  • Sialic Acid Binding Immunoglobulin-like Lectins* / genetics
  • Sialic Acid Binding Immunoglobulin-like Lectins* / immunology
  • Sialic Acid Binding Immunoglobulin-like Lectins* / metabolism
  • Tumor Microenvironment / immunology

Substances

  • Sialic Acid Binding Immunoglobulin-like Lectins
  • Receptors, Chimeric Antigen
  • SIGLEC9 protein, human
  • Antigens, CD