PMID- 35484716 OWN - NLM STAT- MEDLINE DCOM- 20220624 LR - 20220716 IS - 2198-3844 (Electronic) IS - 2198-3844 (Linking) VI - 9 IP - 18 DP - 2022 Jun TI - Single-cell RNA Sequencing Identified Novel Nr4a1(+) Ear2(+) Anti-Inflammatory Macrophage Phenotype under Myeloid-TLR4 Dependent Regulation in Anti-Glomerular Basement Membrane (GBM) Crescentic Glomerulonephritis (cGN). PG - e2200668 LID - 10.1002/advs.202200668 [doi] LID - 2200668 AB - Previously, this study demonstrates the critical role of myeloid specific TLR4 in macrophage-mediated progressive renal injury in anti-glomerular basement membrane (anti-GBM) crescentic glomerulonephritis (cGN); however, the underlying mechanism remains largely unknown. In this study, single-cell RNA sequencing (scRNA-seq), pseudotime trajectories reconstruction, and motif enrichment analysis are used, and macrophage diversity in anti-GBM cGN under tight regulation of myeloid-TLR4 is uncovered. Most significantly, a myeloid-TLR4 deletion-induced novel reparative macrophage phenotype (Nr4a1(+) Ear2+) with significant upregulated anti-inflammatory and tissue repair-related signaling is discovered, thereby suppressing the M1 proinflammatory responses in anti-GBM cGN. This is further demonstrated in vitro that deletion of TLR4 from bone marrow-derived macrophages (BMDMs) induces the Nr4a1/Ear2-expressing anti-inflammatory macrophages while blocking LPS-stimulated M1 proinflammatory responses. Mechanistically, activation of the Nr4a1/Ear2-axis is recognized as a key mechanism through which deletion of myeloid-TLR4 promotes the anti-inflammatory macrophage differentiation in vivo and in vitro. This is confirmed by specifically silencing macrophage Nr4a1 or Ear2 to reverse the anti-inflammatory effects on TLR4 deficient BMDMs upon LPS stimulation. In conclusion, the findings decode a previously unidentified role for a myeloid-TLR4 dependent Nr4a1/Ear2 negative feedback mechanism in macrophage-mediated progressive renal injury, implying that activation of Nr4a1-Ear2 axis can be a novel and effective immunotherapy for anti-GBM cGN. CI - (c) 2022 The Authors. Advanced Science published by Wiley-VCH GmbH. FAU - Chen, Jiaoyi AU - Chen J AUID- ORCID: 0000-0001-5601-9411 AD - Department of Medicine and Therapeutics, Li Ka Shing Institute of Health Sciences, and Lui Che Woo Institute of Innovative Medicine, The Chinese University of Hong Kong, Hong Kong, 999077, P. R. China. FAU - Huang, Xiao Ru AU - Huang XR AD - Department of Medicine and Therapeutics, Li Ka Shing Institute of Health Sciences, and Lui Che Woo Institute of Innovative Medicine, The Chinese University of Hong Kong, Hong Kong, 999077, P. R. China. AD - Guangdong-Hong Kong Joint Laboratory on Immunological and Genetic Kidney Diseases, Guangdong Academy of Medical Sciences, Guangdong Provincial People's Hospital, Guangzhou, 510080, P. R. China. FAU - Yang, Fuye AU - Yang F AD - Department of Medicine and Therapeutics, Li Ka Shing Institute of Health Sciences, and Lui Che Woo Institute of Innovative Medicine, The Chinese University of Hong Kong, Hong Kong, 999077, P. R. China. AD - Department of Nephrology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang, 31009, P. R. China. FAU - Yiu, Wai Han AU - Yiu WH AD - Division of Nephrology, Department of Medicine, The University of Hong Kong, Hong Kong, 999077, P. R. China. FAU - Yu, Xueqing AU - Yu X AD - Guangdong-Hong Kong Joint Laboratory on Immunological and Genetic Kidney Diseases, Guangdong Academy of Medical Sciences, Guangdong Provincial People's Hospital, Guangzhou, 510080, P. R. China. FAU - Tang, Sydney C W AU - Tang SCW AD - Division of Nephrology, Department of Medicine, The University of Hong Kong, Hong Kong, 999077, P. R. China. FAU - Lan, Hui Yao AU - Lan HY AUID- ORCID: 0000-0003-4283-9755 AD - Department of Medicine and Therapeutics, Li Ka Shing Institute of Health Sciences, and Lui Che Woo Institute of Innovative Medicine, The Chinese University of Hong Kong, Hong Kong, 999077, P. R. China. AD - The Chinese University of Hong Kong-Guangdong Academy of Sciences/Guangdong Provincial People's Hospital Joint Research Laboratory on Immunological and Genetic Kidney Diseases, The Chinese University of Hong Kong, Hong Kong, 999077, P. R. China. LA - eng GR - 14117418/Research Grants Council of Hong Kong/ GR - 14104019/Research Grants Council of Hong Kong/ GR - 14101121/Research Grants Council of Hong Kong/ GR - R4012-18/Research Grants Council of Hong Kong/ GR - C7018-16G/Research Grants Council of Hong Kong/ GR - 2019B121205005/Guangdong-Hong Kong-Macao-Joint Labs Program from Guangdong Science and Technology Department/ GR - Lui Che Woo Institute of Innovative Medicine/ PT - Journal Article DEP - 20220428 PL - Germany TA - Adv Sci (Weinh) JT - Advanced science (Weinheim, Baden-Wurttemberg, Germany) JID - 101664569 RN - 0 (Anti-Inflammatory Agents) RN - 0 (Lipopolysaccharides) RN - 0 (NR4A1 protein, human) RN - 0 (Nuclear Receptor Subfamily 4, Group A, Member 1) RN - 0 (TLR4 protein, human) RN - 0 (Toll-Like Receptor 4) RN - EC 3.1.- (Eosinophil-Derived Neurotoxin) SB - IM MH - Anti-Inflammatory Agents MH - Eosinophil-Derived Neurotoxin/metabolism MH - Glomerular Basement Membrane MH - *Glomerulonephritis/genetics MH - Humans MH - Lipopolysaccharides MH - Macrophages MH - Nuclear Receptor Subfamily 4, Group A, Member 1 MH - Phenotype MH - Sequence Analysis, RNA MH - *Toll-Like Receptor 4 PMC - PMC9218767 OTO - NOTNLM OT - Nr4a1/Ear2 OT - anti-GBM crescentic glomerulonephritis OT - macrophages OT - myeloid-TLR4 COIS- The authors declare no conflict of interest. EDAT- 2022/04/30 06:00 MHDA- 2022/06/25 06:00 PMCR- 2022/04/28 CRDT- 2022/04/29 00:23 PHST- 2022/03/07 00:00 [revised] PHST- 2022/02/03 00:00 [received] PHST- 2022/04/30 06:00 [pubmed] PHST- 2022/06/25 06:00 [medline] PHST- 2022/04/29 00:23 [entrez] PHST- 2022/04/28 00:00 [pmc-release] AID - ADVS3938 [pii] AID - 10.1002/advs.202200668 [doi] PST - ppublish SO - Adv Sci (Weinh). 2022 Jun;9(18):e2200668. doi: 10.1002/advs.202200668. Epub 2022 Apr 28.