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Identification of HuR-specific RNA interactome in TGF-ß1-activated fibroblasts in response to cigarette smoke

Sofia Winslow, Llywelyn Griffith, Henric Olsson, Jernej Ule, Zala Jevnikar Rojnik
European Respiratory Journal 2021 58: PA2397; DOI: 10.1183/13993003.congress-2021.PA2397
Sofia Winslow
1Translational Science and Experimental Medicine, Research and Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden
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  • For correspondence: sofia.winslow@astrazeneca.com
Llywelyn Griffith
2The Francis Crick Institute, London, United Kingdom
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Henric Olsson
1Translational Science and Experimental Medicine, Research and Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden
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Jernej Ule
2The Francis Crick Institute, London, United Kingdom
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Zala Jevnikar Rojnik
1Translational Science and Experimental Medicine, Research and Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden
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Abstract

Aims and objectives: Smoking is a risk factor for idiopathic pulmonary fibrosis (IPF), but the associated mechanism remains unknown. Our aim was to study the RNA-regulatory networks of the stress-responsive RNA-binding protein HuR and what impact it has on the disease pathology.

Methods: TGF-β1-activated human lung fibroblasts were used to study the effect of cigarette smoke extract (CSE) stimulation on the translocation HuR from the nucleus to the cytoplasm. HuR-specific RNA-interactions in the nuclear and cytoplasmic cell fractions were analyzed by CLIP (cross-linking and immunoprecipitation) and subsequent high-throughput sequencing.

Results: We showed that CSE induces HuR translocation from the nucleus to the cytoplasm in TGF-β1-activated lung fibroblasts. CLIP data revealed several fibrotic genes, including COL1A1 and COL1A2, among the most frequently bound HuR-targets in the cytoplasm. Evaluation of HuR-bound RNA sequences showed a difference in 4-meric binding motifs between nuclear and cytoplasmic HuR fractions, with increased binding to polyU-rich RNA motifs in the cytoplasm. Interestingly, the polyU-rich binding was reduced upon TGF-β1/CSE stimulation. Additionally, the TGF-β1/CSE stimulation led to reduced binding of cytoplasmic HuR to 3’UTR RNA regions and increased interaction with the coding regions.

Conclusions: Our data suggests a global shift of HuR function from 3’UTR-dependent RNA stabilization, potentially in the direction of translational regulation of genes associated with fibrosis.

  • Idiopathic pulmonary fibrosis
  • Smoking

Footnotes

Cite this article as: European Respiratory Journal 2021; 58: Suppl. 65, PA2397.

This abstract was presented at the 2021 ERS International Congress, in session “Prediction of exacerbations in patients with COPD”.

This is an ERS International Congress abstract. No full-text version is available. Further material to accompany this abstract may be available at www.ers-education.org (ERS member access only).

  • Copyright ©the authors 2021
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Identification of HuR-specific RNA interactome in TGF-ß1-activated fibroblasts in response to cigarette smoke
Sofia Winslow, Llywelyn Griffith, Henric Olsson, Jernej Ule, Zala Jevnikar Rojnik
European Respiratory Journal Sep 2021, 58 (suppl 65) PA2397; DOI: 10.1183/13993003.congress-2021.PA2397

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Identification of HuR-specific RNA interactome in TGF-ß1-activated fibroblasts in response to cigarette smoke
Sofia Winslow, Llywelyn Griffith, Henric Olsson, Jernej Ule, Zala Jevnikar Rojnik
European Respiratory Journal Sep 2021, 58 (suppl 65) PA2397; DOI: 10.1183/13993003.congress-2021.PA2397
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