Project description: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.
Project description:To study the safety and efficacy of clinical application of autologous tumor infiltrating lymphocytes (TIL) after in vitro amplification，in combination with anti-PD-1 monoclonal antibody for the treatment of advanced hepatocellular carcinoma (HCC)
Project description:Fresh human HCC tissue and paired pertumor liver tissue were acquired, isolation of the infiltrating lymphocytes was conducted by discontinous density gradient centrifugation in Percoll. After removal of dead cells and incubation with components of TCRγ/δ isolation kit (Mitenyi Biotec), γδ T cells were positively selected from the single-cell suspension using autoMACS Separator (Miltenyi Biotec) with recommended programs. Then we employed whole genome microarray expression profiling as a discovery platform to indentify genes which are differentially expressed between HCC-derived and paired peritumor-derived γδT cells.
Project description:Study goal is to disclose features of gene expressio profile of non-cancerous liver-infiltrating lymphocytes of type C hepatitis patients with hepatocellular carcinomas and tumor-infiltrating lymphocytes of type C hepatitis patients with hepatocellular carcinomas. Keywords: gene expression profile, non-cancerous liver-infiltrating lymphocytes, tumor-infiltrating lymphocytes, type C hepatitis, hepatocellular carcinoma Non-cancerous liver-infiltrating lymphocytes were obtained by laser capture microdissection from surgically resected liver tissues of 12 type C hepatitis patients with hepatocellular carcinoma. The mRNA was amplified and expression profile was comprehensively analyzed with reference RNA using oligo-DNA chip. Tumor-infiltrating lymphocytes were obtained by laser capture microdissection from surgically resected cancer tissues of 12 type C hepatitis patients with hepatocellular carcinoma. The mRNA was amplified and expression profile was comprehensively analyzed with reference RNA using oligo-DNA chip.
Project description:We found that the number of tumor-infiltrating myofibroblasts was positively correlated to tumor acidification status in hepatocellular carcinoma (HCC). Hepatic stellate cells (HSCs), the predominant precursors of liver myofibroblasts, were activated and transdifferentiated into myofibroblasts under acidic culture condition. To identify the molecular phenotype of LX-2 cells in acidic culture conditions, we further conducted a gene expression profile analysis. LX-2 cells cultured in pH 7.2 or pH 6.2 medium separately for six days was used in gene expression microarray analysis.
