Urine-derived exosomes from individuals with IPF carry pro-fibrotic cargo

Abstract

Background: MicroRNAs (miRNA) and other components contained in extracellular vesicles may reflect the presence of a disease. Lung tissue, sputum and sera of individuals with idiopathic pulmonary fibrosis (IPF) show alterations in miRNA expression. We designed this study to test whether urine and/or tissue derived exosomal miRNAs from individuals with IPF carry cargo that can promote fibrosis. Methods: Exosomes were isolated from urine (U-IPFexo), lung tissue myofibroblasts (MF-IPFexo), serum from individuals with IPF (n=16) and age/sex-matched controls without lung disease (n=10). We analyzed microRNA expression of isolated exosomes and their in vivo bio-distribution. We investigated the effect on ex vivo skin wound healing and in in vivo mouse lung models. Results: U-IPFexo or MF-IPFexo expressed miR let-7d, miR-29a-5p, miR 181b-3p and miR-199a-3p consistent with previous reports of miRNA expression obtained from lung tissue/sera from patients with IPF. In vivo bio-distribution experiments detected bioluminescent exosomes in the lung of normal C57Bl6 mice within 5 minutes after intravenous infusion, followed by distribution to other organs irrespective of exosome source. Exosomes labeled with gold nanoparticles and imaged by transmission electron microscopy were visualized in alveolar epithelial type I and type II cells. Treatment of human and mouse lung punches obtained from control, non-fibrotic lungs with either U-IPFexo or MF-IPFexo produced a fibrotic phenotype. A fibrotic phenotype was also induced in a human ex vivo skin model and in in vivo lung models. Conclusions: Our results provide evidence of a systemic feature of IPF whereby exosomes contain pro-fibrotic miRNAs when obtained from a fibrotic source and interfere with response to tissue injury as measured in skin and lung models. Funding: This work was supported in part by Lester and Sue Smith Foundation and The Samrick Family Foundation and NIH grants R21 AG060338 (SE and MKG), U01 DK119085 (IP, RS, MTC).

Fibroblasts-derived extracellular vesicles contain SFRP1 and mediate pulmonary fibrosis

“…Whether the level of vesicular SFRP1 correlates with fibrosis severity remains an open question which will be addressed in followup studies. In line, EVs were also found in urine of patients with IPF (Elliot et al, 2022). The authors reported a specific miRNA composition but little is known about the proteins transported by these vesicles.…”

“…We and others demonstrated the crucial role of EVs during pulmonary fibrosis (Martin-Medina et al, 2018; Njock et al, 2019; Parimon et al, 2019; Santos-Alvarez et al, 2022). These vesicles accumulate within the lungs of patients with IPF and can be collected in several body fluids such as BALF, sputum or urine (Elliot et al, 2022; Martin-Medina et al, 2018; Njock et al, 2019). During fibrosis, EVs exacerbate major mechanisms of disease such as proliferation and activation of fibroblasts (Elliot et al, 2022; Martin-Medina et al, 2018) or AT2 cells (Parimon et al, 2019).…”

“…These vesicles accumulate within the lungs of patients with IPF and can be collected in several body fluids such as BALF, sputum or urine (Elliot et al, 2022; Martin-Medina et al, 2018; Njock et al, 2019). During fibrosis, EVs exacerbate major mechanisms of disease such as proliferation and activation of fibroblasts (Elliot et al, 2022; Martin-Medina et al, 2018) or AT2 cells (Parimon et al, 2019). Besides, EVs may also have anti-fibrotic properties via the inhibition of TGF-β signaling (Guiot et al, 2020).…”