Dataset Information

Glutamine deprivation impairs function of infiltrating CD8⁺ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis.

ABSTRACT:

Background

The functions of infiltrating CD8⁺ T cells are often impaired due to tumor cells causing nutrient deprivation in the tumor microenvironment. Thus, the mechanisms of CD8⁺ T cell dysfunction have become a hot research topic, and there is increased interest on how changes in metabolomics correlate with CD8⁺ T cell dysfunction.

Aim

To investigate whether and how glutamine metabolism affects the function of infiltrating CD8⁺ T cells in hepatocellular carcinoma.

Methods

Immunohistochemical staining and immunofluorescence were performed on surgically resected liver tissues from patients. Differentially expressed genes in infiltrating CD8⁺ T cells in hepatocellular carcinoma were detected using RNA sequencing. Activated CD8⁺ T cells were co-cultured with Huh-7 cells for 3 d. The function and mitochondrial status of CD8⁺ T cells were analyzed by flow cytometry, quantitative real-time polymerase chain reaction, and transmission electron microscopy. Next, CD8⁺ T cells were treated with the mitochondrial protective and damaging agents. Functional alterations in CD8⁺ T cells were detected by flow cytometry. Then, complete medium without glutamine was used to culture cells and their functional changes and mitochondrial status were detected.

Results

There were a large number of infiltrating PD-1⁺CD8⁺ T cells in liver cancer tissues. Next, we co-cultured CD8⁺ T cells and Huh-7 cells to explore the regulatory effect of hepatoma cells on CD8⁺ T cells. Flow cytometry results revealed increased PD-1 expression and decreased secretion of perforin (PRF1) and granzyme B (GZMB) by CD8⁺ T cells in the co-culture group. Meanwhile, JC-1 staining was decreased and the levels of reactive oxygen species and apoptosis were increased in CD8⁺ T cells of the co-culture group; additionally, the mitochondria of these cells were swollen. When CD8⁺ T cells were treated with the mitochondrial protective and damaging agents, their function was restored and inhibited, respectively, through the mitochondrial damage and apoptotic pathways. Subsequently, complete medium without glutamine was used to culture cells. As expected, CD8⁺ T cells showed functional downregulation, mitochondrial damage, and apoptosis.

Conclusion

Glutamine deprivation impairs the function of infiltrating CD8⁺ T cells in hepatocellular carcinoma through the mitochondrial damage and apoptotic pathways.

SUBMITTER: Wang W

PROVIDER: S-EPMC9244988 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Glutamine deprivation impairs function of infiltrating CD8<sup>+</sup> T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis.

Wang Wei W Guo Meng-Nan MN Li Ning N Pang De-Quan DQ Wu Jing-Hua JH

World journal of gastrointestinal oncology 20220601 6

<h4>Background</h4>The functions of infiltrating CD8<sup>+</sup> T cells are often impaired due to tumor cells causing nutrient deprivation in the tumor microenvironment. Thus, the mechanisms of CD8<sup>+</sup> T cell dysfunction have become a hot research topic, and there is increased interest on how changes in metabolomics correlate with CD8<sup>+</sup> T cell dysfunction.<h4>Aim</h4>To investigate whether and how glutamine metabolism affects the function of infiltrating CD8<sup>+</sup> T cell ...[more]

PMID: 35949216

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Project description:Background and aimsTargeting costimulatory receptors with agonistic antibodies is a promising cancer immunotherapy option. We aimed to investigate costimulatory receptor expression, particularly 4-1BB (CD137 or tumor necrosis factor receptor superfamily member 9), on tumor-infiltrating CD8+ T cells (CD8+ tumor-infiltrating lymphocytes [TILs]) and its association with distinct T-cell activation features among exhausted CD8+ TILs in hepatocellular carcinoma (HCC).Approach and resultsTumor tissues, adjacent nontumor tissues, and peripheral blood were collected from HCC patients undergoing surgical resection (n = 79). Lymphocytes were isolated and used for multicolor flow cytometry, RNA-sequencing, and in vitro functional restoration assays. Among the examined costimulatory receptors, 4-1BB was most prominently expressed on CD8+ TILs. 4-1BB expression was almost exclusively detected on CD8+ T cells in the tumor-especially on programmed death 1 (PD-1)high cells and not PD-1int and PD-1neg cells. Compared to PD-1int and 4-1BBneg PD-1high CD8+ TILs, 4-1BBpos PD-1high CD8+ TILs exhibited higher levels of tumor reactivity and T-cell activation markers and significant enrichment for T-cell activation gene signatures. Per-patient analysis revealed positive correlations between percentages of 4-1BBpos cells among CD8+ TILs and levels of parameters of tumor reactivity and T-cell activation. Among highly exhausted PD-1high CD8+ TILs, 4-1BBpos cells harbored higher proportions of cells with proliferative and reinvigoration potential. Our 4-1BB-related gene signature predicted survival outcomes of HCC patients in the The Cancer Genome Atlas cohort. 4-1BB agonistic antibodies enhanced the function of CD8+ TILs and further enhanced the anti-PD-1-mediated reinvigoration of CD8+ TILs, especially in cases showing high levels of T-cell activation.Conclusion4-1BB expression on CD8+ TILs represents a distinct activation state among highly exhausted CD8+ T cells in HCC. 4-1BB costimulation with agonistic antibodies may be a promising strategy for treating HCCs exhibiting prominent T-cell activation.

Project description:BackgroundGastric cancer (GC) is a leading malignant disease in numerous countries, including Taiwan with limited therapeutic options. Animal viruses including oncolytic avian reovirus (ARV) have the possibility to avoid pre-existing immunity in humans, while being safe and immunostimulatory. Here, we provide a novel insight into oncolytic ARV and UV-ARV-sensitized patient's peripheral blood mononuclear cells (P-PBMCs) and tumor infiltrating lymphocytes (TILs) killing primary GC (PGC) cells through the surface TLR3 and TRAIL/DR4/DR5 immunogenic apoptosis pathway.MethodsWe conducted a comprehensive study to reveal whether ARV- or UV-inactivated ARV (UV-ARV)-modulated P-PBMCs or TILs killing ARV- and UV-ARV-sensitized AGS cells and PGC cells derived from clinical patients and to investigate the regulation of surface TLR3 receptor and upstream signaling pathways. Apoptosis analysis by flow cytometry and Western blot, suppression of signal pathway by specific inhibitors, in situ proximity ligation assay (PLA), time-resolved flurometry and lactate dehydrogenase (LDH) cytotoxicity assays, and an in vitro co-culture model were established to study the interplay between ARV- and UV-ARV-sensitized P-PBMCs and TILs to kill PGC cells and their upstream pathways.ResultsOur results reveal that increased levels of DR4 and DR5 were observed in ARV and UV-ARV sensitized PGC cells through the TLR3/p38/p53 signaling pathway. Importantly, we found that the σC protein of ARV or UV-ARV interacted with surface TLR3 of CD8+ TILs, thereby triggering the TLR3/NF-κB/IFN-γ/TRAIL signaling pathway which induces immunogenic apoptosis of PGC cells. This study sheds further light on the molecular basis behind ARV oncolysis and facilitates the ARV or UV-ARV as a cancer therapeutic.ConclusionsThe study provides novel insights into ARV- or UV-ARV-sensitized P-PBMCs and CD8+ TILs to kill PGC cells through the immunogenic apoptosis pathway. We conclude that P-PBMCs can easily be obtained from GC patients and provide a rich source as TILs to kill PGC cells.

Dataset Information

Glutamine deprivation impairs function of infiltrating CD8⁺ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis.

Background

Aim

Methods

Results

Conclusion

Publications

Glutamine deprivation impairs function of infiltrating CD8<sup>+</sup> T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis.

Background

Aim

Methods

Results

Conclusion

Publications

Glutamine deprivation impairs function of infiltrating CD8<sup>+</sup> T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Glutamine deprivation impairs function of infiltrating CD8⁺ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis.