Single-cell RNA-seq analysis reveals BHLHE40-driven pro-tumour neutrophils with hyperactivated glycolysis in pancreatic tumour microenvironment
L Wang, Y Liu, Y Dai, X Tang, T Yin, C Wang, T Wang… - Gut, 2023 - gut.bmj.com
L Wang, Y Liu, Y Dai, X Tang, T Yin, C Wang, T Wang, L Dong, M Shi, J Qin, M Xue, Y Cao…
Gut, 2023•gut.bmj.comObjective Innate immunity plays important roles in pancreatic ductal adenocarcinoma
(PDAC), as non-T-cell-enriched tumour. Neutrophils are major players in innate immune
system. Here, we aimed to explore the heterogeneity and pro-tumour mechanisms of
neutrophils in PDAC. Design We analysed single-cell transcriptomes of peripheral blood
polymorphonuclear leucocytes (PMNs) and tumour-infiltrating immune cells from five
patients with PDAC, and performed immunofluorescence/immunohistochemistry staining …
(PDAC), as non-T-cell-enriched tumour. Neutrophils are major players in innate immune
system. Here, we aimed to explore the heterogeneity and pro-tumour mechanisms of
neutrophils in PDAC. Design We analysed single-cell transcriptomes of peripheral blood
polymorphonuclear leucocytes (PMNs) and tumour-infiltrating immune cells from five
patients with PDAC, and performed immunofluorescence/immunohistochemistry staining …
Objective
Innate immunity plays important roles in pancreatic ductal adenocarcinoma (PDAC), as non-T-cell-enriched tumour. Neutrophils are major players in innate immune system. Here, we aimed to explore the heterogeneity and pro-tumour mechanisms of neutrophils in PDAC.
Design
We analysed single-cell transcriptomes of peripheral blood polymorphonuclear leucocytes (PMNs) and tumour-infiltrating immune cells from five patients with PDAC, and performed immunofluorescence/immunohistochemistry staining, multi-omics analysis and in vitro experiments to validate the discoveries of bioinformatics analysis.
Results
Exploration of the heterogeneity of tumour-associated neutrophils (TANs) revealed a terminally differentiated pro-tumour subpopulation (TAN-1) associated with poor prognosis, an inflammatory subpopulation (TAN-2), a population of transitional stage that have just migrated to tumour microenvironment (TAN-3) and a subpopulation preferentially expressing interferon-stimulated genes (TAN-4). Glycolysis signature was upregulated along neutrophil transition trajectory, and TAN-1 was featured with hyperactivated glycolytic activity. The glycolytic switch of TANs was validated by integrative multi-omics approach of transcriptomics, proteomics and metabolomics analysis. Activation of glycolytic activity by LDHA overexpression induced immunosuppression and pro-tumour functions in neutrophil-like differentiated HL-60 (dHL-60) cells. Mechanistic studies revealed BHLHE40, downstream to hypoxia and endoplasmic reticulum stress, was a key regulator in polarisation of neutrophils towards TAN-1 phenotype, and direct transcriptional regulation of BHLHE40 on TAN-1 marker genes was demonstrated by chromatin immunoprecipitation assay. Pro-tumour and immunosuppression functions were observed in dHL-60 cells overexpressing BHLHE40. Importantly, immunohistochemistry analysis of PDAC tissues revealed the unfavourable prognostic value of BHLHE40+ neutrophils.
Conclusion
The dynamic properties of TANs revealed by this study will be helpful in advancing PDAC therapy targeting innate immunity.
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