Fibroblast-derived extracellular vesicles contain SFRP1 and mediate pulmonary fibrosis
Olivier Burgy, … , Gerald Burgstaller, Melanie Königshoff
Olivier Burgy, … , Gerald Burgstaller, Melanie Königshoff
Published September 24, 2024
Citation Information: JCI Insight. 2024;9(18):e168889. https://doi.org/10.1172/jci.insight.168889.
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Research Article Cell biology Pulmonology

Fibroblast-derived extracellular vesicles contain SFRP1 and mediate pulmonary fibrosis

Abstract

Idiopathic pulmonary fibrosis (IPF) is a lethal chronic lung disease characterized by aberrant intercellular communication, extracellular matrix deposition, and destruction of functional lung tissue. While extracellular vesicles (EVs) accumulate in the IPF lung, their cargo and biological effects remain unclear. We interrogated the proteome of EV and non-EV fractions during pulmonary fibrosis and characterized their contribution to fibrosis. EVs accumulated 14 days after bleomycin challenge, correlating with decreased lung function and initiated fibrogenesis in healthy precision-cut lung slices. Label-free proteomics of bronchoalveolar lavage fluid EVs (BALF-EVs) collected from mice challenged with bleomycin or control identified 107 proteins enriched in fibrotic vesicles. Multiomic analysis revealed fibroblasts as a major cellular source of BALF-EV cargo, which was enriched in secreted frizzled related protein 1 (SFRP1). Sfrp1 deficiency inhibited the activity of fibroblast-derived EVs to potentiate lung fibrosis in vivo. SFRP1 led to increased transitional cell markers, such as keratin 8, and WNT/β-catenin signaling in primary alveolar type 2 cells. SFRP1 was expressed within the IPF lung and localized at the surface of EVs from patient-derived fibroblasts and BALF. Our work reveals altered EV protein cargo in fibrotic EVs promoting fibrogenesis and identifies fibroblast-derived vesicular SFRP1 as a fibrotic mediator and potential therapeutic target for IPF.

Authors

Olivier Burgy, Christoph H. Mayr, Déborah Schenesse, Efthymios Fousekis Papakonstantinou, Beatriz Ballester, Arunima Sengupta, Yixin She, Qianjiang Hu, Maria Camila Melo-Narvaéz, Eshita Jain, Jeanine C. Pestoni, Molly Mozurak, Adriana Estrada-Bernal, Ugochi Onwuka, Christina Coughlan, Tanyalak Parimon, Peter Chen, Thomas Heimerl, Gert Bange, Bernd T. Schmeck, Michael Lindner, Anne Hilgendorff, Clemens Ruppert, Andreas Güenther, Matthias Mann, Ali Önder Yildirim, Oliver Eickelberg, Anna Lena Jung, Herbert B. Schiller, Mareike Lehmann, Gerald Burgstaller, Melanie Königshoff

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Figure 2

Label-free proteomics identifies several proteins specific to fibrotic EVs.

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Label-free proteomics identifies several proteins specific to fibrotic E...
(A) C57BL/6J mice were exposed to orotracheal bleomycin or NaCl as control. At 14 days after injection, lung function was assessed and BALF was collected. BALF was utilized for EV isolation, and vesicular (EV pellet) as well as nonvesicular counterparts (EV-free SN) were subjected to label-free proteomics (n = 8 for control, n = 6 for bleomycin). (B) Principal component analysis representation of the different samples. (C) Heatmap of the identified proteins. Color coding corresponds to z-scored MS intensity values after imputation. Based on the unsupervised clustering, proteins were grouped into 7 clusters called A to G. (D) Levels of nidogen-1 (Western blot) in EVs from bleomycin or control mice are shown. TSG-101 was used to show protein enrichment in EVs, and protein content for each sample is shown with Ponceau. A549 lysate served as positive control. Equal (10 μg) protein content was used for the Western blot. (E) AGER and EGFR levels assessed by ELISA on normal (blue) and fibrotic (red) EVs. Data are presented as analyte concentration (pg/mL) normalized to 2 × 108 vesicles. Statistical analysis by nonparametric Mann-Whitney. SN, EV-free fraction.