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Reprogrammed lung epithelial cells by decrease of miR-451a in extracellular vesicles contribute to aggravation of pulmonary fibrosis

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Abstract

Extracellular vesicles (EVs) play novel roles in homeostasis through cell-to-cell communication in human airways via transferring miRNAs. However, the contribution of EV miRNAs to pulmonary phenotypic homeostasis is not clearly understood. Hence, the aim of this study was to elucidate the functional role of miRNAs obtained from epithelium-derived EVs in lung fibrogenesis. Pulmonary fibrosis was induced by exposure of polyhexamethylene guanidine phosphate (PHMG-p)-instilled mice. In histopathological changes, a clear phenotypic change was observed in bronchial epithelium. For figuring out the role of EVs derived from conditioned media of untreated cells (EV-Con) and PHMG-p-treated BEAS-2B (EV-PHMG), significant increase in EVs released from PHMG-p-treated BEAS-2B was detected. Functional analysis with targets of differentially expressed miRNAs in EVs was annotated to epithelial–mesenchymal transition (EMT). Especially, the most abundant miRNA, miR-451a, was downregulated in EV of PHMG-p-treated BEAS-2B cells. We found that odd-skipped related 1 (OSR1) was a putative target for miR-451a, which had been known as a transcription factor of several fibrosis-associated genes. Transfer of decreased miR-451a via EV-PHMG upregulated OSR1 and induced EMT compared to Con-EV-treated cells. In pulmonary fibrosis mice, miR-451a levels were significantly reduced in EV derived from bronchoalveolar lavage fluid and OSR1 expression was increased in lung tissues of mice with PHMG-p exposure. MiR-451a-transfected EVs markedly alleviated fibrogenesis in the PHMG-p-exposed lungs. Low level of miR-451a in EVs modulated EMT and fibrogenesis in recipient cells by increasing OSR1 levels in vitro and in vivo. Our results suggest that transferring EV miR-451a induces anti-fibrotic autocrine effect by downregulating its target, OSR1 maintaining pulmonary homeostasis disrupted by PHMG-p exposure, which can be a potential therapeutic target.

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Funding

This work was supported by the Global Ph.D. Fellowship Program (NRF-2016H1A2A1908513) and Basic Science Research Program (NRF-2017R1D1A1B03036438) funded by the Ministry of Education, and the Korea Environmental Industry and Technology Institute (grant number 2018002490005).

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Author notes

  1. Mi Ho Jeong and Ha Ryong Kim are equal contributors.

Authors and Affiliations

  1. Center for Systems Biology, Massachusetts General Hospital, Boston, MA, 02114, USA

    Mi Ho Jeong

  2. College of Pharmacy, Daegu Catholic University, Gyeongsangbuk-do 38430, Gyeongsan, Republic of Korea

    Ha Ryong Kim

  3. School of Pharmacy, Sungkyunkwan University, Seobu-ro, Jangan-gu, Gyeonggi-do 16419, 2066, Suwon-si, Republic of Korea

    Yong Joo Park, Kyu Hyuck Chung & Hyung Sik Kim

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  1. Mi Ho Jeong
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  2. Ha Ryong Kim
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Contributions

Mi Ho Jeong designed and performed the experiments, and wrote the manuscript; Yong Joo Park analyzed the data; and Ha Ryong Kim, Hyung Sik Kim, and Kyu Hyuck Chung conceived the design and idea of the study.

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Correspondence to Kyu Hyuck Chung or Hyung Sik Kim.

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Research ethics approval

All animal experiments were approved by the Sungkyunkwan University Animal Care Committee (SKKUIACUC2019-07–09-2), and were conducted in accordance with the guidelines of the National Institutes of Health.

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Highlights

• PHMG-p, a biocide, decreases EV miR-451a significantly in conditioned media of lung epithelial cells.

• Decreased EV miR-451a upregulated its target, OSR1 in recipient cells, resulting in epithelial-mesenchymal transition.

• In in vivo, decreased EV miR-451a of BALF from PHMG-p-exposed mice contributes the upregulation of OSR1 and pulmonary fibrogenesis.

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Jeong, M.H., Kim, H.R., Park, Y.J. et al. Reprogrammed lung epithelial cells by decrease of miR-451a in extracellular vesicles contribute to aggravation of pulmonary fibrosis. Cell Biol Toxicol 38, 725–740 (2022). https://doi.org/10.1007/s10565-021-09626-9

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  • DOI: https://doi.org/10.1007/s10565-021-09626-9

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