Clathrin-independent endocytosis and retrograde transport in cancer cells promote cytotoxic CD8 T cell activation

Clathrin-independent endocytosis (CIE) mediates the cellular uptake of endogenous and exogenous cargoes, including bacterial toxins and viruses. Endophilin A3-mediated endocytosis is a specific CIE mechanism that differs from fast endophilin-mediated endocytosis (FEME), with ALCAM and L1CAM being the first confirmed Endophilin A3-specific cargoes. Here, we report ICAM1 as a new Endophilin A3-dependent endocytic cargo. ALCAM and ICAM1 are important components of immune synapses (IS), which are polarized structures formed between immune cells and target cells, such as cancer cells. These molecules transduce essential co-stimulatory signals to T cells to help their effective activation and proliferation. We find that both ALCAM and ICAM1 serve as cargoes for retromer-dependent retrograde transport to the trans -Golgi network (TGN) in cancer cells. Interestingly, disrupting Endophilin A3-mediated endocytosis or retromer-dependent retrograde transport machinery impairs activation of autologous cytotoxic CD8 T cells, likely by affecting the polarized redistribution of immune synapse components at the plasma membrane. Altogether, our findings demonstrate that CIE and retrograde transport are key pathways in cancer cells that promote the activation of cytotoxic CD8 T cells.