N-糖基化 LTβR 通过阻断 RORC 泛素化和 FOXP3 转录取增加肝癌中 Th17/Treg 细胞比例

LT R-overexpressing CAR-T cells have demonstrated surprising effectiveness against solid tumors, exhibiting strong anti-exhaustion and proliferation capabilities.

However, the role of LT R in CD4 + T cell differentiation and anti-tumor activity remains unclear. In this study, we employed primary or subcutaneous mouse hepatocellular carcinoma (HCC) models and flow cytometry to study the impact of conditional knock-in of Ltbr on CD4 + T cell differentiation and response, particularly the Th17/Treg cell ratio, and its influence on HCC progression.

Immunoprecipitation, immunoblotting, RT-qPCR, molecular docking, and Chromatin Immunoprecipitation-qPCR were performed to investigate the molecular mechanism of CD4 + T cell differentiation. Adeno-associated virus-modified T cells were introduced into patient-derived orthotopic xenograft (PDOX) model to assess the combined impact of LT R and glycolysis inhibitors on the Th17/Treg cell differentiation.

We found that LT R reduced PELI1 expression, preventing TRAF3 protein degradation in Th17 cells. TRAF3 then competed with RORC for SMURF1 binding, enhancing RORC stability and Th17 cell differentiation. LT R also blocked PRDM1 expression, delaying Foxp3 transcription and Treg cell infiltration.

Additionally, N-glycosylation supported the stability of LT R by protecting it from ubiquitination. From a therapeutic perspective, glycolysis inhibitors helped LT R balance the proportion of Th17/Treg cells in PDOX model to inhibit tumor growth.

In conclusion, our findings indicated that LT R N-glycosylation prevented RORC ubiquitination and Foxp3 transcription, raising the Th17/Treg cell ratio and hindering HCC progression.