Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients

ABSTRACT Vitronectin (VTN) is a potential non‐invasive biomarker for renal fibrosis, originally described in urinary extracellular vesicles (uEV) from kidney transplant patients (KTx). However, VTN's specific role in renal fibrosis is unclear, as it is involved in various physiological processe...

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Main Authors: Marta Clos‐Sansalvador, Sergio G. Garcia, Paula Rodríguez‐Martínez, Marta Sanroque‐Muñoz, Miriam Font‐Morón, Cristina Grange, Benedetta Bussolati, Marcella Franquesa, Javier Juega, Francesc E. Borràs
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Language:English
Published: Wiley 2025-06-01
Series:Journal of Extracellular Biology
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Online Access:https://doi.org/10.1002/jex2.70056
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author Marta Clos‐Sansalvador
Sergio G. Garcia
Paula Rodríguez‐Martínez
Marta Sanroque‐Muñoz
Miriam Font‐Morón
Cristina Grange
Benedetta Bussolati
Marcella Franquesa
Javier Juega
Francesc E. Borràs
author_facet Marta Clos‐Sansalvador
Sergio G. Garcia
Paula Rodríguez‐Martínez
Marta Sanroque‐Muñoz
Miriam Font‐Morón
Cristina Grange
Benedetta Bussolati
Marcella Franquesa
Javier Juega
Francesc E. Borràs
author_sort Marta Clos‐Sansalvador
collection DOAJ
description ABSTRACT Vitronectin (VTN) is a potential non‐invasive biomarker for renal fibrosis, originally described in urinary extracellular vesicles (uEV) from kidney transplant patients (KTx). However, VTN's specific role in renal fibrosis is unclear, as it is involved in various physiological processes. This study aims to identify other uEV‐associated proteins linked to renal fibrosis to clarify which pathways involve VTN. uEV were isolated from 33 KTx patients and five healthy controls. uEV proteins were analysed using proximity extension assay (PEA), and data were normalized and compared using Welch's two‐sided t‐test to identify differentially expressed proteins between fibrotic (n = 31) and non‐fibrotic patients (n = 7). Urinary VTN levels and monocyte chemoattractant protein‐1 (MCP‐1) were measured by ELISA. PEA analysis identified 33 proteins overexpressed in the fibrotic group. These proteins clustered in STRING analysis, primarily associating with coagulation, fibrinolysis and TNF‐inflammation involving macrophages. ELISA detection of MCP‐1 further validated the results. High levels of VTN in the fibrotic group were accompanied by the upregulation of fibrinolytic pathway components (PAI‐1, tPA and uPAR), which are well‐known to interact with VTN. This study highlights TNF‐induced inflammation involving macrophages and fibrinolysis as key mechanisms underlying renal fibrosis with direct implications of VTN, which support VTN's potential as a biomarker for this pathological process.
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spelling doaj-art-7144e055f3d141ca9a68512e6580e4e42025-08-20T03:27:10ZengWileyJournal of Extracellular Biology2768-28112025-06-0146n/an/a10.1002/jex2.70056Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant PatientsMarta Clos‐Sansalvador0Sergio G. Garcia1Paula Rodríguez‐Martínez2Marta Sanroque‐Muñoz3Miriam Font‐Morón4Cristina Grange5Benedetta Bussolati6Marcella Franquesa7Javier Juega8Francesc E. Borràs9REMAR‐IGTP Group Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP), Can Ruti Campus Badalona (Barcelona) Catalonia SpainREMAR‐IGTP Group Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP), Can Ruti Campus Badalona (Barcelona) Catalonia SpainDepartment of Pathology University Hospital Germans Trias i Pujol Badalona (Barcelona) SpainREMAR‐IGTP Group Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP), Can Ruti Campus Badalona (Barcelona) Catalonia SpainREMAR‐IGTP Group Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP), Can Ruti Campus Badalona (Barcelona) Catalonia SpainUniversity of Turin: Universita degli Studi di Torino Turin ItalyUniversity of Turin: Universita degli Studi di Torino Turin ItalyREMAR‐IGTP Group Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP), Can Ruti Campus Badalona (Barcelona) Catalonia SpainREMAR‐IGTP Group Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP), Can Ruti Campus Badalona (Barcelona) Catalonia SpainREMAR‐IGTP Group Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP), Can Ruti Campus Badalona (Barcelona) Catalonia SpainABSTRACT Vitronectin (VTN) is a potential non‐invasive biomarker for renal fibrosis, originally described in urinary extracellular vesicles (uEV) from kidney transplant patients (KTx). However, VTN's specific role in renal fibrosis is unclear, as it is involved in various physiological processes. This study aims to identify other uEV‐associated proteins linked to renal fibrosis to clarify which pathways involve VTN. uEV were isolated from 33 KTx patients and five healthy controls. uEV proteins were analysed using proximity extension assay (PEA), and data were normalized and compared using Welch's two‐sided t‐test to identify differentially expressed proteins between fibrotic (n = 31) and non‐fibrotic patients (n = 7). Urinary VTN levels and monocyte chemoattractant protein‐1 (MCP‐1) were measured by ELISA. PEA analysis identified 33 proteins overexpressed in the fibrotic group. These proteins clustered in STRING analysis, primarily associating with coagulation, fibrinolysis and TNF‐inflammation involving macrophages. ELISA detection of MCP‐1 further validated the results. High levels of VTN in the fibrotic group were accompanied by the upregulation of fibrinolytic pathway components (PAI‐1, tPA and uPAR), which are well‐known to interact with VTN. This study highlights TNF‐induced inflammation involving macrophages and fibrinolysis as key mechanisms underlying renal fibrosis with direct implications of VTN, which support VTN's potential as a biomarker for this pathological process.https://doi.org/10.1002/jex2.70056fibrinolysisproximity extension assayrenal fibrosisTNFurinary extracellular vesiclesvitronectin
spellingShingle Marta Clos‐Sansalvador
Sergio G. Garcia
Paula Rodríguez‐Martínez
Marta Sanroque‐Muñoz
Miriam Font‐Morón
Cristina Grange
Benedetta Bussolati
Marcella Franquesa
Javier Juega
Francesc E. Borràs
Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients
Journal of Extracellular Biology
fibrinolysis
proximity extension assay
renal fibrosis
TNF
urinary extracellular vesicles
vitronectin
title Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients
title_full Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients
title_fullStr Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients
title_full_unstemmed Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients
title_short Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients
title_sort proteomics of urinary extracellular vesicles highlight the involvement of vitronectin and the fibrinolytic and tnf pathways as mechanisms underlying renal fibrosis in kidney transplant patients
topic fibrinolysis
proximity extension assay
renal fibrosis
TNF
urinary extracellular vesicles
vitronectin
url https://doi.org/10.1002/jex2.70056
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