Li et al. J Transl Med (2020) 18:92 https://doi.org/10.1186/s12967-020-02267-2 Journal of Translational Medicine RESEARCH Open Access Comprehensive analysis of the association between tumor glycolysis and immune/ infammation function in breast cancer Wenhui Li1†, Ming Xu1†, Yu Li1, Ziwei Huang1, Jun Zhou1, Qiuyang Zhao1, Kehao Le1, Fang Dong1, Cheng Wan2 and Pengfei Yi1* Abstract Background: Metabolic reprogramming, immune evasion and tumor-promoting infammation are three hallmarks of cancer that provide new perspectives for understanding the biology of cancer. We aimed to fgure out the relation- ship of tumor glycolysis and immune/infammation function in the context of breast cancer, which is signifcant for deeper understanding of the biology, treatment and prognosis of breast cancer. Methods: Using mRNA transcriptome data, tumor-infltrating lymphocytes (TILs) maps based on digitized H&E- stained images and clinical information of breast cancer from The Cancer Genome Atlas projects (TCGA), we explored the expression and prognostic implications of glycolysis-related genes, as well as the enrichment scores and dual role of diferent immune/infammation cells in the tumor microenvironment. The relationship between glycolysis activity and immune/infammation function was studied by using the diferential genes expression analysis, gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, gene set enrichment analyses (GSEA) and correlation analysis. Results: Most glycolysis-related genes had higher expression in breast cancer compared to normal tissue. Higher phosphoglycerate kinase 1 (PGK1) expression was associated with poor prognosis. High glycolysis group had upregu- lated immune/infammation-related genes expression, upregulated immune/infammation pathways especially IL-17 signaling pathway, higher enrichment of multiple immune/infammation cells such as Th2 cells and macrophages. However, high glycolysis group was associated with lower infltration of tumor-killing immune cells such as NKT cells and higher immune checkpoints expression such as PD-L1, CTLA4, FOXP3 and IDO1. Conclusions: In conclusion, the enhanced glycolysis activity of breast cancer was associated with pro-tumor immu- nity. The interaction between tumor glycolysis and immune/infammation function may be mediated through IL-17 signaling pathway. Keywords: Breast cancer, Glycolysis, Immune/infammation function, PGK1, IL-17 signaling pathway, TCGA, Prognosis Background Breast cancer is the leading cause of cancer death among women [1]. Breast cancer can be classifed as Nor- *Correspondence: [email protected] †Wenhui Li and Ming Xu contributed equally to this work mal-like, Luminal A, Luminal B, HER2-enriched and 1 Department of Breast and Thyroid Surgery, Union Hospital, Tongji Basal-like subtypes. In addition to surgery, systemic ther- Medical College, Huazhong University of Science and Technology, apies including chemotherapy, hormonal therapy, and Wuhan 430022, China Full list of author information is available at the end of the article molecular targeted therapy can be chosen based on the © The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creat iveco mmons .org/licen ses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creat iveco mmons .org/publi cdoma in/ zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Li et al. J Transl Med (2020) 18:92 Page 2 of 12 molecular characteristics to combat cancer [2]. Although selection. So far, however, there has been little compre- these systemic therapies have improved patients’ out- hensive analysis focusing on the relationships between comes, many patients do not respond to these existing the tumor glycolysis, immune/infammation function treatments, which leads to poor prognosis [3, 4]. On and the clinical features based on clinical data in the feld this account, some researches have been carried out to of breast cancer. Given this, we implemented studies with explore new efective therapies in breast cancer such as transcriptome and clinical data of breast cancer from Te Immunotherapy and metabolic therapy [5, 6]. However, Cancer Genome Atlas (TCGA) projects to explore the only a minority of patients beneft from these emerg- landscape of tumor glycolysis and immunity in breast ing therapies [7]. Exploring the interplay between tumor cancer, to identify the relationship between the tumor cells and tumor microenvironment could lead to deeper glycolysis and immune and infammatory cells infltra- understanding of breast cancer initiation, progression, tion, and to fgure out the impact of the two on breast and therapeutic resistance, possibly provide prospective cancer prognosis. strategies for cancer prevention and treatment [8, 9]. Metabolic reprogramming is a key hallmark of cancer Methods [10]. Te most frequently mentioned way of metabolic The preprocessing step for transcriptome and clinical data reprogramming is aerobic glycolysis. Aerobic glycolysis, We obtained mRNA transcriptome data of breast cancer also known as the “Warburg efect”, is a general way of from TCGA. Te RNA sequence data of breast cancer glucose metabolism in cancer cells. In this way, glucose is was downloaded directly from TCGA data portal (https mainly processed into lactate even when oxidative capac- ://porta l.gdc.cance r.gov/). Te obtained expression data ity is intact. Tis will lead to a highly acidic microenvi- of 19657 genes from 1208 samples including 112 normal ronment. According to current research, tumor aerobic tissues and 1096 tumor tissues were used for follow-up glycolysis can contribute to malignant transformation research. Te corresponding clinicopathological features and tumor progression [11]. Terefore, tumor aerobic including ER, PR, Her2 status, tumor stage, survival time glycolysis has possible implications for prognosis judg- and survival status were downloaded from TCGA data ment and cancer treatment [12]. Exploiting tumor gly- portal. colysis for clinical application requires fguring out how intrinsic and extrinsic factors to be integrated to modify Identifcation of diferentially expressed genes the metabolic phenotype [13]. and functional enrichment analysis Tumor microenvironment (TME), cancer cells’ sup- Te tumor samples were divided into low glycoly- porting hotbed, has complicated and changeable compo- sis group and high glycolysis group based on the median sition including tumor infltrating lymphocytes (TILs), expression of PGK1. Gene expression profles were other immune and infammatory cells, fbroblasts, the derived based on diferential genes expression analysis blood and lymphatic vascular networks, the extracellular using limma R package. Diferentially expressed genes matrix (ECM) and so on [14]. Numerous evidences sug- (DEGs) were identifed through volcano plot fltering. gest that the immune cells infltration in the TME could Te thresholds for DEGs were |log2FC| ≥ 1 and adjusted interact with tumor cells, which may afect tumor pro- P value < 0.05. Gene ontology (GO) analysis and Kyoto gression and the efcacy of existing anticancer therapies Encyclopedia of Genes and Genomes (KEGG) analysis [15, 16]. Te immune cells recruited to the tumor site were performed using DAVID (https ://david .ncifc rf.gov/) have dual characters, some can restrain carcinogenesis and WebGestalt (http://www.webge stalt .org/) [23, 24], and tumor progression while others may play a tumor- using P < 0.05 as the cut-of criterion. Gene set enrich- promoting role [17]. Tus, it is important to fgure out ment analysis (GSEA) was conducted to fnd out the the cellular heterogeneity composition of the immune signifcantly upregulated and downregulated pathways cells infltration and the cause behind it. Te results may between high and low glycolysis group using WebGestalt, be signifcant for optimizing existing treatments and using FDR < 0.05 as the cut-of criterion. identifying novel therapeutic targets [18]. Several studies have investigated the relationship Cell type enrichment analysis between tumor glycolysis and immune/infammation To evaluate the cellular heterogeneity of the tumor function [19–21]. A highly acidic microenvironment microenvironment, the xCell tool, a gene signature-based due to tumor glycolysis may diferentially infuence method, was used to fgure out the abundance of cell immune cells infltration, ultimately leading to immune types in tumor microenvironment, based on the tissue escape and cancer progression [22]. Exploration of the transcriptome profles [25]. Total 64 cell types including associations is providing a deeper look into cancer bio- adaptive and innate immune cells, stem cells, epithelial logical processes and can lead to more efective therapy