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Identification of transforming growth factorbeta-lregulated microRNAs and the microRNAtargetomes in primary lung fibroblasts

Ong, J., Timens, W., Rajendran, V., Algra, A., Spira, A., Lenburg, M. E., Campbell, J. D., van den Berge, M., Postma, D. S., van den Berg, A., Kluiver, J. & Brandsma, C-A., 14-Sep-2017, In : PLoS ONE. 12, 9, 24 p., e0183815.

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BACKGROUND: Lung fibroblasts are involved in extracellular matrix homeostasis, which is mainly regulated by transforming growth factor-beta (TGF-β), and are therefore crucial in lung tissue repair and remodeling. Abnormal repair and remodeling has been observed in lung diseases like COPD. As miRNA levels can be influenced by TGF-β, we hypothesized that TGF-β influences miRNA expression in lung fibroblasts, thereby affecting their function.

MATERIALS AND METHODS: We investigated TGF-β1-induced miRNA expression changes in 9 control primary parenchymal lung fibroblasts using miRNA arrays. TGF-β1-induced miRNA expression changes were validated and replicated in an independent set of lung fibroblasts composted of 10 controls and 15 COPD patients using qRT-PCR. Ago2-immunoprecipitation followed by mRNA expression profiling was used to identify the miRNA-targetomes of unstimulated and TGF-β1-stimulated primary lung fibroblasts (n = 2). The genes affected by TGF-β1-modulated miRNAs were identified by comparing the miRNA targetomes of unstimulated and TGF-β1-stimulated fibroblasts.

RESULTS: Twenty-nine miRNAs were significantly differentially expressed after TGF-β1 stimulation (FDR<0.05). The TGF-β1-induced miR-455-3p and miR-21-3p expression changes were validated and replicated, with in addition, lower miR-455-3p levels in COPD (p<0.05). We identified 964 and 945 genes in the miRNA-targetomes of unstimulated and TGF-β1-stimulated lung fibroblasts, respectively. The TGF-β and Wnt pathways were significantly enriched among the Ago2-IP enriched and predicted targets of miR-455-3p and miR-21-3p. The miR-455-3p target genes HN1, NGF, STRADB, DLD and ANO3 and the miR-21-3p target genes HHEX, CHORDC1 and ZBTB49 were consistently more enriched after TGF-β1 stimulation.

CONCLUSION: Two miRNAs, miR-455-3p and miR-21-3p, were induced by TGF-β1 in lung fibroblasts. The significant Ago2-IP enrichment of targets of these miRNAs related to the TGF-β and/or Wnt pathways (NGF, DLD, HHEX) in TGF-β1-stimulated fibroblasts suggest a role for these miRNAs in lung diseases by affecting lung fibroblast function.

Original languageEnglish
Article numbere0183815
Number of pages24
JournalPLoS ONE
Volume12
Issue number9
Publication statusPublished - 14-Sep-2017

    Keywords

  • OBSTRUCTIVE PULMONARY-DISEASE, GENE-EXPRESSION, TISSUE-REPAIR, COPD, FIBROSIS, CANCER, INHIBITION, TARGETS, AIRWAY, CELLS

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