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Inhibition of Glycolysis in Pathogenic TH17 Cells Through Targeting a -21Mir −Peli1− C-Rel Pathway Prevents Autoimmunity

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Inhibition of Glycolysis in Pathogenic TH17 Cells Through Targeting a -21Mir −Peli1− C-Rel Pathway Prevents Autoimmunity Inhibition of Glycolysis in Pathogenic TH17 Cells through Targeting a miR-21−Peli1− c-Rel Pathway Prevents Autoimmunity This information is current as Rong Qiu, Xiang Yu, Li Wang, Zhijun Han, Chao Yao, of September 26, 2021. Yange Cui, Guojun Hou, Dai Dai, Wenfei Jin and Nan Shen J Immunol published online 15 May 2020 http://www.jimmunol.org/content/early/2020/05/14/jimmun ol.2000060 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2020/05/14/jimmunol.200006 Material 0.DCSupplemental http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 26, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2020 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published May 15, 2020, doi:10.4049/jimmunol.2000060 The Journal of Immunology Inhibition of Glycolysis in Pathogenic TH17 Cells through Targeting a miR-21–Peli1–c-Rel Pathway Prevents Autoimmunity Rong Qiu,*,†,1 Xiang Yu,*,1 Li Wang,* Zhijun Han,‡ Chao Yao,† Yange Cui,† Guojun Hou,* Dai Dai,* Wenfei Jin,‡ and Nan Shen*,x,{,||,# It is well known that some pathogenic cells have enhanced glycolysis; the regulatory network leading to increased glycolysis are not well characterized. In this study, we show that CNS-infiltrated pathogenic TH17 cells from diseased mice specifically upregulate glycolytic pathway genes compared with homeostatic intestinal TH17 cells. Bioenergetic assay and metabolomics analyses indicate that in vitro–derived pathogenic TH17 cells are highly glycolytic compared with nonpathogenic TH17 cells. Chromatin landscape analyses demonstrate TH17 cells in vivo that show distinct chromatin states, and pathogenic TH17 cells show enhanced chromatin Downloaded from accessibility at glycolytic genes with NF-kB binding sites. Mechanistic studies reveal that miR-21 targets the E3 ubiquitin ligase Peli1–c-Rel pathway to promote glucose metabolism of pathogenic TH17 cells. Therapeutic targeting c-Rel–mediated glycolysis in pathogenic TH17 cells represses autoimmune diseases. These findings extend our understanding of the regulation TH17 cell glycolysis in vivo and provide insights for future therapeutic intervention to TH17 cell–mediated autoimmune diseases. The Journal of Immunology, 2020, 204: 000–000. http://www.jimmunol.org/ + subset of effector CD4 TH cells that produce IL-17A lamina propria (3, 4). Under autoimmune or pathogen infection (1, 2), TH17 cells, mediates barrier tissue integrity and condition, TH17 cells acquire the capability to produce IFN-g A mucosal defense and contributes to the development and GM-CSF and are often seen at sites of inflamed loci (5). of multiple autoimmune diseases. At steady-state condition, TH17 cells also could be differentiated in vitro by a combi- TH17 cells producing IL-10 are mainly found at sites of intestinal nation of cytokines, either by TGF-b1plusIL-6(TH17 [b], nonpathogenic) or by IL-6 plus IL-1b and IL-23 (TH17 [23], pathogenic) (6, 7). *Shanghai Institute of Rheumatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200001, China; †Shanghai Institute of Nutrition and Upon Ag stimulation, lymphocytes undergo extensive clonal ex- Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy pansion and differentiation for immune defense or tolerance (8–11). by guest on September 26, 2021 ‡ of Sciences, Chinese Academy of Sciences, Shanghai 200001, China; Department of Activated lymphocytes are highly glycolytic and demonstrate a Biology, Southern University of Science and Technology, Shenzhen 518000, China; xShenzhen Futian Hospital for Rheumatic Diseases, Shenzhen 518000, China; {State striking increase in cell content and nutrients uptake. c-Myc, c-Rel, Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji and mTORc1-dependent glycolytic activity is crucial for effective T Hospital, Shanghai 200001, China; ||Center for Autoimmune Genomics and Etiology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229; and #Department and B cell immune response (12–15), whereas Foxo1 and Foxp3 of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229 proteins function as key metabolic repressors that limit glycolytic 1R.Q. and X.Y. contributed equally. activity in regulatory T cells (16, 17). It was reported that by ORCIDs: 0000-0001-7395-0419 (L.W.); 0000-0001-9374-2510 (C.Y.); 0000-0002- restricting glucose availability, tumor cells limit aerobic glycolysis 0028-3739 (W.J.). and effector function of tumor-infiltrating T cells (18, 19). Recently, it Received for publication January 21, 2020. Accepted for publication April 1, 2020. was demonstrated that TH17 cells developed in the presence of seg- This work was supported by grants from the National Natural Science Foundation of mented filamentous bacteria or Citrobacter rodentium show func- China (31630021, 81421001, and 31930037), the Shanghai Municipal Key Medical Center tional and metabolic heterogeneity, with Citro rodentium–induced Construction Project (2017ZZ01024-002), the Shanghai Sailing Program (17YF1417900), the Shenzhen Science and Technology Project (JCYJ20180504170414637), and the pathogenic TH17 cells showing a global upregulation of multiple Sanming Project of Medicine in Shenzhen (SZSM201602087). This work was done metabolic pathways (20). by an innovative research team of high-level local universities in Shanghai led by N.S. Tight communication between epigenetic remodeling and meta- R.Q., X.Y., W.J., and N.S. designed the experiments and wrote the manuscript; R.Q. bolic regulation for cell plasticity is beginning to be understood and X.Y. did most of the experiments; L.W. helped with the mouse experiments; Z.H. and C.Y. helped with ATAC-seq and RNA-seq data analysis; Y.C., D.D., and G.H. (21, 22). Assay for transposase-accessible chromatin technol- helped with RT-PCR experiments. ogy sequencing (ATAC-seq) was developed to study global The sequences presented in this article have been submitted to the Gene Expression chromatin dynamics by using relatively low cell number (23). It Omnibus (https://www.ncbi.nlm.nih.gov/geo/) under accession number GSE127768. was successfully used to study chromatin dynamics of regula- Address correspondence and reprint requests to Prof. Nan Shen, Shanghai Institute of tory T cells under inflammatory condition (24), development of Rheumatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, 145 plasmacytoid dendritic cells (25), and tumor-specific CD8+ Shan Dong Road, Shanghai 200001, China. E-mail address: [email protected] T cells (26). However, chromatin architecture of T 17 cells The online version of this article contains supplemental material. H in vivo is unknown, and signaling pathways that induce TH17 Abbreviations used in this article: ATAC-seq, assay for transposase-accessible chro- matin technology sequencing; EAE, experimental autoimmune encephalomyelitis; cell chromatin remodeling are also incompletely understood. ECAR, extracellular acidification rate; FC, fold change; GC-TOFMS, gas chromato- microRNAs are a class of noncoding RNAs that modulate gene graph time-of-flight mass spectrometry; PTXF, pentoxifylline; RIP, RNA-binding expression at the posttranscriptional level (27). Specific micro- protein immunoprecipitation; RNA-seq, RNA sequencing. RNAs have been reported to be essential for the metabolic func- Copyright Ó 2020 by The American Association of Immunologists, Inc. 0022-1767/20/$37.50 tion of certain cells such as miR-33a/b, which was shown to be www.jimmunol.org/cgi/doi/10.4049/jimmunol.2000060 2 miR-21 CONTROLS GLYCOLYSIS OF PATHOGENIC TH17 CELLS crucial for cholesterol and lipid metabolism (28, 29), and let-7 Intracellular cytokine staining family members were recently shown to be essential for B cell Cultured or tissue isolated lymphocytes were washed and stimulated with metabolic function (30). The regulation of TH17 cell metabolic PMA (50 ng/ml) plus ionomycin (750 ng/ml) for 4 h at 37˚C. Cells were reprogramming in vivo to distinct microenvironment by micro- stained with LIVE/DEAD BV510 (catalog no. L34955; Thermo Fisher RNAs is not well understood. Scientific), anti-CD45 APC-Cy7 (catalog no. 557659; BD Biosciences), In this study, first by bioenergetic and metabolomics study, we anti-CD3 PerCP5.5 (catalog no. 35-0031-82; eBioscience), and anti-CD4 BV421 (catalog no. 562891; BD Biosciences) for 30 min at 4˚C. Cells demonstrate that in vitro–derived nonpathogenic TH17 (b) cells were then fixed, permeabilized with Perm/Wash buffer (catalog no. show reduced glycolytic activity compared with pathogenic TH17 555028; BD Biosciences), and stained with anti–IFN-g APC (catalog no. (23) cells. Further, by global transcriptional analyses, we dem- 554413; BD Biosciences), anti–GM-CSF PE (catalog no. 554406; BD onstrate that homeostatic T
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