Inhibition of glycosphingolipid synthesis overcomes the steric hindrance of CD30 N-glycans to augment CD30-targeted immunotherapeutic efficacy.

CD30-targeted chimeric antigen receptor (CAR) T-cell therapy faces clinical challenges in classical Hodgkin lymphoma (cHL). While current optimization strategies focus on CAR design, manufacturing protocols, and preconditioning regimens, tumor-intrinsic resistance mechanisms remain poorly understood. Our study revealed that CD30 in cHL cells is associated with N-glycans at Asn101 and Asn276, which are essential for protein stability but do not affect cell proliferation or apoptosis. Genetic ablation of these N-glycans or enzymatic deglycosylation significantly enhanced CD30-targeted CAR-T-cell accessibility to tumor cells, leading to improved T-cell activation and cytotoxic function. Notably, pretreatment with eliglustat, an FDA-approved glycosphingolipid synthesis inhibitor, selectively potentiated the antitumor activity of CD30-targeted CAR-T cells in wild-type CD30-expressing tumors but had minimal effects on CD30 glycosylation-deficient variants. Eliglustat combined with CD30-targeted CAR-T cells resulted in superior tumor control in xenograft models without additional toxicity. Mechanistically, eliglustat trimmed terminal sialic acids from CD30 N-glycans while preserving the core N-glycan structure. Furthermore, the addition of eliglustat also enhanced the tumor-killing activity of brentuximab vedotin (BV), a CD30-directed antibody-drug conjugate, both in vitro and in vivo. This glycoimmunotherapy paradigm represents a clinically actionable approach to overcome glycan-mediated immune evasion and enhance therapeutic efficacy in CD30-positive lymphomas.