Differential expression of insulin-like growth factor type 1 receptor identifies heterogeneous intrahepatic regulatory T subsets in mouse hepatocellular carcinoma

Understanding regulatory T-cell (Treg)-mediated tumor tolerance is critical for designing immunotherapy against hepatocellular carcinoma (HCC). In this study, we characterized the expression of insulin-like growth factor type 1 receptor (IGF1R) in intrahepatic Tregs in a chemical-induced mouse HCC model. We found two intrahepatic Treg subsets with differential IGF1R expression: IGF1Rhi Tregs and IGF1Rlo/– Tregs. Functional assays indicated that compared with IGF1Rlo/– Tregs, IGF1Rhi Tregs produced more TGF-β and IL-10 and were more proliferative in vivo. Furthermore, IGF1Rhi Tregs exhibited higher phosphorylation of the mammalian target of the rapamycin complex 1 (mTORC1) in vivo. However, in vitro stimulation and immunosuppression assay revealed that the immunosuppressive capacity of the two Treg subsets was equivalent, as evidenced by comparable cytokine production and immunosuppressive effect over conventional T cells. The transcriptome sequencing analysis revealed up-regulation of genes that encode proteins essential for glycolysis, oxidative phosphorylation, and electron transport chain in IGF1Rhi Tregs. Consistently, IGF1Rhi Tregs produces more adenosine triphosphate (ATP), lactate, and reactive oxygen species (ROS). Furthermore, malignant cells in the tumor nodules induced IGF1R down-regulation in Tregs at the mRNA level. In summary, we identified the heterogeneity of intrahepatic Tregs in HCC which might play significant roles in tumor immunity.