CELL INTELLIGENCE · 肿瘤细胞治疗研究

BACKGROUND: Recent studies have shown that CD32/CD8a/CD28/CD3 chimeric receptor cells directly kill breast cancer cells, suggesting the existence of cell surface myeloid Fc R alternative ligands (ALs). Here, we investigated the metabolism, ALs, cytotoxicity, and immunoregulatory functions of CD64/CD28/CD3 in colorectal cancer (CRC) and squamous cell carcinoma of the head and neck. METHODS: The CD64/CD28/CD3 -SFG retroviral vector was used to produce viruses for T-cell transduction. T-cell expansion and differentiation were monitored via flow cytometry. Gene expression was assessed by RNA-seq. Bioenergetics were documented on a Seahorse extracellular flux analyzer. CD64/CD28/CD3 polarization was identified via confocal microscopy. Cytotoxicity was determined by MTT assay and bioluminescent imaging, and flow cytometry. Tridimensional antitumor activity of CD64/CD28/CD3 T cells was achieved by utilizing HCT116-GFP 3D spheroids via the IncuCyte S3 Live-Cell Analysis system. The intraperitoneal distribution and antitumor activity of NIR-CD64/CD28/CD3 and NIR-nontransduced T cells were investigated in CB17-SCID mice bearing subcutaneous FaDu Luc + cells by bioluminescent and fluorescent imaging. IFN was assessed by ELISA. RESULTS: Compared to CD16/CD8a/CD28/CD3 T cells, CD32/CD8a/CD28/CD3 T cells, and non-transduced T cells, CD64/CD28/CD3 T cells exhibited the highest levels of cell expansion and persistence capacity. A total of 235 genes linked to cell division and 52 genes related to glycolysis were overexpressed. The glycolytic phenotype was confirmed by functional in vitro studies accompanied by preferential T-cell effector memory differentiation. Interestingly, oxamic acid was found to inhibit CD64-CR T cell proliferation, indicating the involvement of lactate. Upon CD64/CD28/CD3 T-cell conjugation with CRC cells, CD64/CD28/CD3 cells polarize at immunological synapses, leading to CRC cell death. CD64/CD28/CD3 T cells kill SCCHN cells, and in combination with the anti-B7-H3 mAb (376.96) or anti-EGFR mAb, these cells trigger antibody-dependent cellular cytotoxicity (ADCC) in vitro under 2D and 3D conditions. The 376.96 mAb combined with CD64/CD28/CD3 T cells had anti-SCCHN activity in vivo. In addition, they induce the upregulation of PD-L1 and HLA-DR expression in cancer cells via IFN . PD-L1 positive SCCHN cells in combination with anti-PD-L1 mAb and CD64-CR T cells were killed by ADCC, which enhanced direct cytotoxicity. These findings indicate that the glycolytic phenotype is involved in CD64-CR T cell proliferation/expansion. These cells mediate long-lasting HLA-independent cytotoxicity and ADCC in CRC and SCCHN cells. CONCLUSIONS: CD64/CD28/CD3 T cells could significantly impact the rational design of personalized studies to treat CRC and SCCHN and the identification of novel Fc R ALs in cancer and healthy cells.