Fingerprints of Hepatocellular Carcinoma-Infiltrated γδT Cells Revealed by Single-Cell Sequencing

We report the application of single cell RNA sequencing for profiling the fingerprints of γδT cells of hepatocellular carcinoma compared with healthy donors. By comparing the transcriptome and TCR clonality of γδT cells from 3 HCC patients and expanded γδT cells from 3 healthy donors, we found that expanded γδ T cells derived from different donors shared great transcriptomic similarity and we found higher enrichments of major γδ TCRs in expanded γδ T cells compared to infiltrated but not peripheral ones in HCC. Analysis of differentiation trajectory demonstrated a branched structure in expanded γδ T cells, implying functional and differentiation stage diversity, and γδ T cells from HCC patients are localized to the far end of the trajectory. Furthermore, GVSA analyses made on metabolic pathways demonstrated strong enrichments in major metabolic pathways such as OXPHOS, glycolysis, and fatty acid metabolism in expanded γδ T cells versus peripheral ones of HCC. The comparison indicated higher cytotoxicity score of expanded γδ T cells compared to infiltrated or peripheral ones of HCC. The successful application of allogeneic γδ T cells immunotherapy in late-stage liver cancer patients was supported by our current scRNA transcriptomic analyses, which suggested strong anti-tumor functional complementation by the ex-vivo expanded γδ T cells. We report the application of single cell RNA sequencing for profiling the fingerprints of γδT cells of hepatocellular carcinoma compared with healthy donors. Transcriptomic and TCR clonality analysis were both applied. The results showed that expanded γδ T cells derived from different donors shared great transcriptomic similarity and higher enrichments of major γδ TCRs in expanded γδ T cells. Analysis of differentiation trajectory demonstrated a branched structure in expanded γδ T cells, implying functional and differentiation stage diversity, and γδ T cells from HCC patients are localized to the far end of the trajectory. Furthermore, GVSA analyses made on metabolic pathways demonstrated strong enrichments in major metabolic pathways such as OXPHOS, glycolysis, and fatty acid metabolism in expanded γδ T cells versus peripheral ones of HCC. The comparison indicated higher cytotoxicity score of expanded γδ T cells compared to infiltrated or peripheral ones of HCC.