Single-cell RNA-seq analysis reveals BHLHE40-driven pro-tumour neutrophils with hyperactivated glycolysis in pancreatic tumour microenvironment

作者：Liwen Wang, Yihao Liu, Yuting Dai, Xiaomei Tang, Yin Tong, Chaofu Wang, Ting Wang, Lei Dong, Minmin Shi, Jiejie Qin, Meilin Xue, Yizhi Cao, Jia Liu, Pengyi Liu, Jinyan Huang, Chenlei Wen, Jun Zhang, Zhiwei Xu, Fan Bai, Xiaxing Deng, Chenghong Peng, Hao Chen, Lingxi Jiang, Sai‐Juan Chen, Baiyong Shen · 发表于：Gut · 年份：2022 · DOI：10.1136/gutjnl-2021-326070 · 被引用次数：330 · 研究领域：Immune cells in cancer、Single-cell and spatial transcriptomics、Phagocytosis and Immune Regulation

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...