The CD64/CD28/CD3ζ chimeric receptor reprograms T-cell metabolism and promotes T-cell persistence and immune functions while triggering antibody-independent and antibody-dependent cytotoxicity

• DOI: 10.1186/s40164-025-00601-2
• Publication Date: 2025-02-18T02:37:51Z
• Authors: Sara Caratelli; Francesca De Paolis; Domenico Alessand...; Silvia Baldari; Illari Salvatori; Apollonia Tullo; Giulia Lanzilli; Aymone Gurtner; Alberto Ferri; Cristiana Valle; Simona Padovani; Valeriana Cesarini; Tommaso Sconocchia; Loredana Cifaldi; Roberto Arriga; Giulio Cesare Spa...; Soldano Ferrone; Adriano Venditti; Piero Rossi; Graziano Pesole; Gabriele Toietta; Giuseppe Sconocchia
• Keywords: Persistence (discontinuity)

Recent studies have shown that CD32/CD8a/CD28/CD3ζ chimeric receptor cells directly kill breast cancer cells, suggesting the existence of cell surface myeloid FcγR alternative ligands (ALs). Here, we investigated metabolism, ALs, cytotoxicity, and immunoregulatory functions CD64/CD28/CD3ζ in colorectal (CRC) squamous carcinoma head neck. The -SFG retroviral vector was used to produce viruses for T-cell transduction. expansion differentiation were monitored via flow cytometry. Gene expression assessed by RNA-seq. Bioenergetics documented on a Seahorse extracellular flux analyzer. polarization identified confocal microscopy. Cytotoxicity determined MTT assay bioluminescent imaging, Tridimensional antitumor activity T achieved utilizing HCT116-GFP 3D spheroids IncuCyte S3 Live-Cell Analysis system. intraperitoneal distribution NIR-CD64/CD28/CD3ζ NIR-nontransduced CB17-SCID mice bearing subcutaneous FaDu Luc + fluorescent imaging. IFNγ ELISA. Compared CD16/CD8a/CD28/CD3ζ non-transduced exhibited highest levels persistence capacity. A total 235 genes linked division 52 related glycolysis overexpressed. glycolytic phenotype confirmed functional vitro accompanied preferential effector memory differentiation. Interestingly, oxamic acid found inhibit CD64-CR proliferation, indicating involvement lactate. Upon conjugation with CRC polarize at immunological synapses, leading death. SCCHN combination anti-B7-H3 mAb (376.96) or anti-EGFR mAb, these trigger antibody-dependent cellular cytotoxicity (ADCC) under 2D conditions. 376.96 combined had anti-SCCHN vivo. In addition, they induce upregulation PD-L1 HLA-DR IFNγ. positive anti-PD-L1 killed ADCC, which enhanced direct cytotoxicity. These findings indicate is involved proliferation/expansion. mediate long-lasting HLA-independent ADCC cells. could significantly impact rational design personalized treat identification novel ALs healthy