PMID- 36726727 OWN - NLM STAT- MEDLINE DCOM- 20230206 LR - 20230427 IS - 2167-8359 (Electronic) IS - 2167-8359 (Linking) VI - 11 DP - 2023 TI - Microscopic polyangiitis plasma-derived exosomal miR-1287-5p induces endothelial inflammatory injury and neutrophil adhesion by targeting CBL. PG - e14579 LID - 10.7717/peerj.14579 [doi] LID - e14579 AB - BACKGROUND: An inflammatory environment around the vessel wall caused by leukocyte infiltration is one of the characteristic histopathological features of microscopic polyangiitis (MPA); however, the pathogenic mechanisms are not fully understood. Studies have found that circulating microRNA (miRNA) can be used as potential biomarkers for the diagnosis and classification of anti-neutrophil cytoplasmic autoantibody (ANCA)-associated vasculitides (AAV), and the E3 ubiquitin ligase casitas B-lineage lymphoma (CBL) seems to be associated with inflammation. In addition, evidence indicates that miRNA can be tracked into exosomes and transferred into recipient cells to mediate the process of vascular endothelial injury. Herein, we aimed to identify the profiles of exosomal miRNA, and determine the effect of exosomal miR-1287-5p and its target gene CBL on vascular endothelial cells in MPA. METHOD: We isolated plasma exosomes from patients with MPA (MPA-exo) and healthy controls (HC-exo) by ultracentrifugation and conducted exosome small-RNA sequencing to screen differential miRNA expression in MPA-exo (n = 3) compared to HC-exo (n = 3). We measured the expression levels of miR-1303, miR-1287-5p, and miR-129-1-3p using quantitative reverse transcription-polymerase chain reaction (qRT-PCR, n = 6) and performed dual luciferase reporter gene assays to confirm the downstream target gene of miR-1287-5p. In addition, we treated human umbilical vein endothelial cell (HUVEC) with MPA-exo, or transfected them with miR-1287-5p mimic/inhibitor or with CBL-siRNA/CBL-siRNA+ miR-1287-5p inhibitor. After cell culture, we evaluated the effects on vascular endothelial cells by examining the mRNA levels of IL-6, IL-8, MCP-1, ICAM-1 and E-selectin using qRT-PCR and performed neutrophil adhesion assay with haematoxylin staining. RESULT: Transmission electron microscopy, Western blot and nanoparticle tracking analysis showed that we successfully purified exosomes and MPA-exo could be absorbed into HUVEC. We screened a total of 1,077 miRNA by sequencing and observed a high abundance of miR-1287-5p in the exosomes obtained from MPA plasma. The dual luciferase reporter assay identified CBL as a downstream target gene of miR-1287-5p, and the results revealed that MPA-exo decreased CBL protein expression in HUVEC. In addition, treatment with MPA-exo, up-regulating miR-1287-5p or silencing of CBL in HUVEC significantly increased the mRNA expression of inflammatory factors (including IL-6, IL-8, and MCP-1) and adhesion molecules (including ICAM-1 and E-selection) and promoted the adhesion of neutrophils to HUVEC. However, down-regulating miR-1287-5p had the opposite effect. CONCLUSION: Our study revealed that MPA-exo was involved in the intercellular transfer of miR-1287-5p and subsequently promote the development of acute endothelial injury in MPA. MiR-1287-5p and CBL agonists may be promising therapeutic approach for MPA-induced vascular inflammatory injury. CI - (c)2023 Zhu et al. FAU - Zhu, Yan AU - Zhu Y AD - Department of Nephrology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China. AD - The First Affiliated Hospital, Department of Nephrology, Hengyang Medical School, University of South China, Hengyang, Hunan, China. FAU - Liu, Liu AU - Liu L AD - Department of Nephrology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China. FAU - Chu, Liepeng AU - Chu L AD - Department of Nephrology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China. FAU - Lan, Jingjing AU - Lan J AD - Department of Nephrology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China. FAU - Wei, Jingsi AU - Wei J AD - Department of Nephrology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China. FAU - Li, Wei AU - Li W AD - Department of Nephrology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China. FAU - Xue, Chao AU - Xue C AD - Department of Nephrology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20230127 PL - United States TA - PeerJ JT - PeerJ JID - 101603425 RN - 126547-89-5 (Intercellular Adhesion Molecule-1) RN - 0 (Interleukin-6) RN - 0 (Interleukin-8) RN - 0 (MicroRNAs) RN - 0 (MIRN1287 microRNA, human) RN - 0 (Mirn129 microRNA, human) RN - 0 (MIRN1303 microRNA, human) RN - 0 (RNA, Messenger) RN - 0 (RNA, Small Interfering) SB - IM MH - Humans MH - Human Umbilical Vein Endothelial Cells/metabolism MH - Intercellular Adhesion Molecule-1/metabolism MH - Interleukin-6/metabolism MH - Interleukin-8/metabolism MH - *MicroRNAs/genetics MH - *Microscopic Polyangiitis/metabolism MH - RNA, Messenger/metabolism MH - RNA, Small Interfering/metabolism MH - Exosomes PMC - PMC9885867 OTO - NOTNLM OT - CBL OT - Exosome OT - Human umbilical vein endothelial cells OT - MiR-1287-5p OT - Microscopic polyangiitis COIS- The authors declare there are no competing interests. EDAT- 2023/02/03 06:00 MHDA- 2023/02/04 06:00 PMCR- 2023/01/27 CRDT- 2023/02/02 02:01 PHST- 2022/03/17 00:00 [received] PHST- 2022/11/28 00:00 [accepted] PHST- 2023/02/02 02:01 [entrez] PHST- 2023/02/03 06:00 [pubmed] PHST- 2023/02/04 06:00 [medline] PHST- 2023/01/27 00:00 [pmc-release] AID - 14579 [pii] AID - 10.7717/peerj.14579 [doi] PST - epublish SO - PeerJ. 2023 Jan 27;11:e14579. doi: 10.7717/peerj.14579. eCollection 2023.