Hsa-miR-21-5p is induced by interleukin-6 and affects multiple pathogenic factors associated with fibroblast-like synoviocytes in rheumatoid arthritis

Abstract Fibroblast-like synoviocytes (FLS) contribute significantly to the pathogenesis of rheumatoid arthritis (RA), particularly through their roles in synovitis and joint destruction. The microRNAs (miRNAs) are short non-coding RNA that regulate gene expression post-transcriptionally. Although m...

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Main Authors: Kei Araki, Sho Mokuda, Hiroki Kohno, Naoya Oka, Hirofumi Watanabe, Michinori Ishitoku, Tomohiro Sugimoto, Yusuke Yoshida, Junya Masumoto, Shintaro Hirata
Format: Article
Language:English
Published: Nature Portfolio 2025-06-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-02840-z
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author Kei Araki
Sho Mokuda
Hiroki Kohno
Naoya Oka
Hirofumi Watanabe
Michinori Ishitoku
Tomohiro Sugimoto
Yusuke Yoshida
Junya Masumoto
Shintaro Hirata
author_facet Kei Araki
Sho Mokuda
Hiroki Kohno
Naoya Oka
Hirofumi Watanabe
Michinori Ishitoku
Tomohiro Sugimoto
Yusuke Yoshida
Junya Masumoto
Shintaro Hirata
author_sort Kei Araki
collection DOAJ
description Abstract Fibroblast-like synoviocytes (FLS) contribute significantly to the pathogenesis of rheumatoid arthritis (RA), particularly through their roles in synovitis and joint destruction. The microRNAs (miRNAs) are short non-coding RNA that regulate gene expression post-transcriptionally. Although more than 2000 human miRNAs are registered, comprehensive analyses of miRNA expression in FLS are limited. Herein, we investigated the relationship between miRNAs and FLS. Next-generation sequencing (small RNA-seq) was performed on primary cultured FLS derived from patients with RA to analyze the mature miRNA expression pattern. To assess inflammation-induced changes in miRNA levels, FLS were cultured with cytokines and evaluated by RT-qPCR. MiRNA mimics were transfected into FLS and an immortalized synovial fibroblast cell line (MH7A cells), and validated using conventional RNA-seq. Out of 2861 mature miRNAs, 297 mature miRNAs were detected and intronic miRNAs were predominated. Notably, hsa-miR-21-5p was abundantly expressed and its expression was enhanced by IL-6 stimulation. The induction of miR-21-5p mimic decreased the expression of Programmed Cell Death 4 (PDCD4) and Osteoprotegerin (OPG), while upregulating Semaphorin 5A (SEMA5A). MiR-21-5p mimic led to enhanced cell proliferation. These data suggest that hsa-miR-21-5p in FLS may exacerbate the pathophysiology of rheumatoid synovitis by promoting FLS proliferation, highlighting its potential as a therapeutic target in RA.
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spelling doaj-art-792551cd32674b08b206cdc391e41e862025-08-20T02:05:13ZengNature PortfolioScientific Reports2045-23222025-06-0115111510.1038/s41598-025-02840-zHsa-miR-21-5p is induced by interleukin-6 and affects multiple pathogenic factors associated with fibroblast-like synoviocytes in rheumatoid arthritisKei Araki0Sho Mokuda1Hiroki Kohno2Naoya Oka3Hirofumi Watanabe4Michinori Ishitoku5Tomohiro Sugimoto6Yusuke Yoshida7Junya Masumoto8Shintaro Hirata9Department of Clinical Immunology and Rheumatology, Hiroshima University HospitalDepartment of Clinical Immunology and Rheumatology, Hiroshima University HospitalDepartment of Clinical Immunology and Rheumatology, Hiroshima University HospitalDepartment of Clinical Immunology and Rheumatology, Hiroshima University HospitalDepartment of Clinical Immunology and Rheumatology, Hiroshima University HospitalDepartment of Clinical Immunology and Rheumatology, Hiroshima University HospitalDepartment of Clinical Immunology and Rheumatology, Hiroshima University HospitalDepartment of Clinical Immunology and Rheumatology, Hiroshima University HospitalDepartment of Pathology, Ehime University Proteo-Science Center and Graduate School of MedicineDepartment of Clinical Immunology and Rheumatology, Hiroshima University HospitalAbstract Fibroblast-like synoviocytes (FLS) contribute significantly to the pathogenesis of rheumatoid arthritis (RA), particularly through their roles in synovitis and joint destruction. The microRNAs (miRNAs) are short non-coding RNA that regulate gene expression post-transcriptionally. Although more than 2000 human miRNAs are registered, comprehensive analyses of miRNA expression in FLS are limited. Herein, we investigated the relationship between miRNAs and FLS. Next-generation sequencing (small RNA-seq) was performed on primary cultured FLS derived from patients with RA to analyze the mature miRNA expression pattern. To assess inflammation-induced changes in miRNA levels, FLS were cultured with cytokines and evaluated by RT-qPCR. MiRNA mimics were transfected into FLS and an immortalized synovial fibroblast cell line (MH7A cells), and validated using conventional RNA-seq. Out of 2861 mature miRNAs, 297 mature miRNAs were detected and intronic miRNAs were predominated. Notably, hsa-miR-21-5p was abundantly expressed and its expression was enhanced by IL-6 stimulation. The induction of miR-21-5p mimic decreased the expression of Programmed Cell Death 4 (PDCD4) and Osteoprotegerin (OPG), while upregulating Semaphorin 5A (SEMA5A). MiR-21-5p mimic led to enhanced cell proliferation. These data suggest that hsa-miR-21-5p in FLS may exacerbate the pathophysiology of rheumatoid synovitis by promoting FLS proliferation, highlighting its potential as a therapeutic target in RA.https://doi.org/10.1038/s41598-025-02840-zRheumatoid arthritisFibroblast-like synoviocytesmicroRNAhsa-miR-21-5pNext-generation sequencing
spellingShingle Kei Araki
Sho Mokuda
Hiroki Kohno
Naoya Oka
Hirofumi Watanabe
Michinori Ishitoku
Tomohiro Sugimoto
Yusuke Yoshida
Junya Masumoto
Shintaro Hirata
Hsa-miR-21-5p is induced by interleukin-6 and affects multiple pathogenic factors associated with fibroblast-like synoviocytes in rheumatoid arthritis
Scientific Reports
Rheumatoid arthritis
Fibroblast-like synoviocytes
microRNA
hsa-miR-21-5p
Next-generation sequencing
title Hsa-miR-21-5p is induced by interleukin-6 and affects multiple pathogenic factors associated with fibroblast-like synoviocytes in rheumatoid arthritis
title_full Hsa-miR-21-5p is induced by interleukin-6 and affects multiple pathogenic factors associated with fibroblast-like synoviocytes in rheumatoid arthritis
title_fullStr Hsa-miR-21-5p is induced by interleukin-6 and affects multiple pathogenic factors associated with fibroblast-like synoviocytes in rheumatoid arthritis
title_full_unstemmed Hsa-miR-21-5p is induced by interleukin-6 and affects multiple pathogenic factors associated with fibroblast-like synoviocytes in rheumatoid arthritis
title_short Hsa-miR-21-5p is induced by interleukin-6 and affects multiple pathogenic factors associated with fibroblast-like synoviocytes in rheumatoid arthritis
title_sort hsa mir 21 5p is induced by interleukin 6 and affects multiple pathogenic factors associated with fibroblast like synoviocytes in rheumatoid arthritis
topic Rheumatoid arthritis
Fibroblast-like synoviocytes
microRNA
hsa-miR-21-5p
Next-generation sequencing
url https://doi.org/10.1038/s41598-025-02840-z
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