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While tumor-associated glycans are known to protect malignant cells from immune attack, the contribution of N-glycans expressed by non-malignant TME populations to CAR-T cell dysfunction remains poorly defined.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We investigated the role of N-glycans in non-malignant TME populations, focusing on M2-like macrophages and hepatic stellate cells in liver metastasis of colorectal (CRC) and pancreatic cancer (PDAC). Using in vitro co-culture systems, transcriptomic analysis, and tumor-bearing humanized mouse models, we assessed how pharmacologic or genetic disruption of key nodes of the N-glycosylation pathway (\n                      <jats:italic>MGAT5<\/jats:italic>\n                      ,\n                      <jats:italic>MAN2A1<\/jats:italic>\n                      and\n                      <jats:italic>ST6GAL1<\/jats:italic>\n                      ) in immune and stromal compartments shapes T-cell function.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      In patient samples, a branched N-glycan signature was associated with transcriptional programs characteristic of tumor-promoting macrophages and stromal cells, linking N-glycosylation to an immunosuppressive TME. Disruption of N-glycan synthesis in non-malignant TME cells reduced their immunosuppressive and tumor-supporting functions. Single-cell RNA sequencing of tumor-bearing humanized mice showed depletion of protumor IL1\u03b2\n                      <jats:sup>+<\/jats:sup>\n                      macrophages and diminished inhibitory macrophage-T cell interactions following N-glycosylation blockade. Selective\n                      <jats:italic>MGAT5<\/jats:italic>\n                      disruption in immune and stromal compartments suppressed immunosuppressive programs and enhanced CAR-T cell antitumor activity independently of tumor cell glycosylation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      These findings show that N-glycans expressed by non-malignant TME cells restrain CAR-T cell responses in CRC and PDAC, highlighting\n                      <jats:italic>MGAT5<\/jats:italic>\n                      -dependent branching as a potentially actionable axis and supporting a broader role for multiple nodes of the N-glycosylation pathway.\n                    <\/jats:p>\n                  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