Rationale Extracellular vesicles (EVs) are small lipid vesicles, and EV-coupled microRNAs (miRNAs) are important modulators of biological processes. Fibrocytes are circulating bone marrow-derived cells that migrate into the injured lungs and contribute to fibrogenesis. The question of whether EV-coupled miRNAs derived from fibrocytes are able to regulate pulmonary fibrosis has not been addressed yet.
Methods Pulmonary fibrosis was induced in rats by intratracheal administration of an adenoviral gene vector encoding active transforming growth factor-β1 (TGF-β1) or control vector. Primary fibrocytes and fibroblasts were cultured from rat lungs and were sorted by anti-CD45 magnetic beads. Human circulating fibrocytes and fibrocytes in bronchoalveolar lavage fluid (BALF) were isolated by fibronectin-coated dishes. Fibrocytes were cultured on different stiffness plates or decellularised lung scaffolds. We also determined the effects of extracellular matrix (ECM) and recombinant TGF-β1 on the cellular and EV-coupled miRNA expression of fibrocytes.
Results The EVs of fibrocytes derived from fibrotic lungs significantly upregulated the expression of col1a1 of fibroblasts. Culturing on rigid plates or fibrotic decellularised lung scaffolds increased miR-21-5 p expression compared with soft plates or normal lung scaffolds. Dissolved ECM collected from fibrotic lungs and recombinant TGF-β1 increased miR-21-5 p expression on fibrocytes, and these effects were attenuated on soft plates. Fibrocytes from BALF collected from fibrotic interstitial pneumonia patients showed higher miR-21-5 p expression than those from other patients.
Conclusions Our results indicate that ECM contributes to fibrogenesis through biomechanical and biochemical effects on miRNA expression in fibrocytes.
- idiopathic pulmonary fibrosis
Data availability statement
All data relevant to the study are included in the article or uploaded as supplementary information.
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Contributors SS, YN and MRJK contributed to conception and design; SS, CU, TY, TS and MRJK contributed to analysis and interpretation; SS, SGC, CU, TY, CS, P-SB and MRJK contributed to drafting the manuscript for important intellectual content; all authors have approved the final version and agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.
Funding SS was funded by MSD Life Science Foundation and Public Interest Incorporated Foundation (no award number), outside the submitted work. Toyoshi Yanagihara was funded by the Uehara Memorial Foundation Research Fellowship (no award number) and Mitacs Canada (IT06744), outside the submitted work.
Competing interests MRJK reports grants and personal fees from the Canadian Institute for Health Research (PJT-162295), Roche, Boehringer Ingelheim, GSK, Gilead, Prometic, Alkermes, Actelion, Respivert and Synairgen.
Provenance and peer review Not commissioned; externally peer reviewed.
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