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Here, we identify ICAM1 as a novel EndoA3-dependent cargo, alongside ALCAM. We demonstrate that both proteins subsequently follow retromer-dependent retrograde transport to the\n                    <jats:italic>trans<\/jats:italic>\n                    -Golgi network (TGN) in cancer cells. From there, we propose that they undergo polarized redistribution to the plasma membrane, where they contribute to immune synapse formation between cancer cells and cytotoxic CD8 T cells. Disruption of EndoA3 or retromer components significantly affects the response of autologous cytotoxic CD8 T cells, as evidenced by reduced cytokine production and secretion, but increased lytic activity, while proliferation and later activation marker expression remain intact. This is accompanied by diminished ICAM1 density at the immune synapse, where we observe it arriving via polarized vesicular transport, indicating altered synapse organization. Indeed, cancer cells lacking EndoA3-mediated CIE or retromer form enlarged immune synapses that fail to sustain full T cell cytokine secretion, suggesting a compensatory attempt by T cells to overcome the defective synapse, while likely promoting more transient contacts that potentially favor serial killing. Together, these findings reveal that EndoA3-mediated CIE and retrograde transport act in concert in cancer cells to relocate immune synapse components via the Golgi, thereby fine-tuning the balance between cytotoxic T cell cytokine secretion and lytic activity. These insights contribute to a better understanding of the mechanisms governing immune synapse formation and organization, providing a necessary foundation for the long-term identification of new strategies to enhance T cell\u2013mediated anti-tumor immunity.\n                  <\/jats:p>","DOI":"10.7554\/elife.105821","type":"journal-article","created":{"date-parts":[[2025,2,17]],"date-time":"2025-02-17T07:25:08Z","timestamp":1739777108000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.105821","source":"Crossref","is-referenced-by-count":0,"title":["Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response"],"prefix":"10.7554","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0696-499X","authenticated-orcid":true,"given":"Shiqiang","family":"Xu","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/038bmak92","id-type":"ROR","asserted-by":"publisher"}],"name":"UNamur, Namur 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