Extracellular vesicles derived from patients with antibody-mediated rejection induce tubular senescence and endothelial to mesenchymal transition in renal cells

Abstract

Extracellular vesicles (EV) are emerging mediators in several diseases. However, their role in the pathophysiology of antibody‐mediated allograft rejection (AMR) has been poorly investigated. Here, we investigated the role of EV isolated from AMR patients in inducing tubular senescence and endothelial to mesenchymal transition (EndMT) and analyzed their miRNA expression profile. By multiplex bead flow cytometry, we characterized the immunophenotype of plasma AMR‐derived EV and found a prevalent platelet and endothelial cell origin. In vitro, AMR‐derived EV induced tubular senescence by upregulating SA‐β Gal and CDKN1A mRNA. Furthermore, AMR‐derived EV induced EndMT. The occurrence of tubular senescence and EndMT was confirmed by analysis of renal biopsies from the same AMR patients. Moreover, AMR‐derived EV induced C3 gene upregulation and CFH downregulation in tubular epithelial cells, with C4d deposition on endothelial cells. Interestingly, RNase‐mediated digestion of EV cargo completely abrogated tubular senescence and EndMT. By microarray analysis, miR‐604, miR‐515‐3p, miR‐let‐7d‐5p, and miR‐590‐3p were significantly upregulated in EV from AMR group compared with transplant controls, whereas miR‐24‐3p and miR‐29a‐3p were downregulated. Therefore, EV‐associated miRNA could act as active player in AMR pathogenesis, unraveling potential mechanisms of accelerated graft senescence, complement activation and early fibrosis that might lead to new therapeutic intervention.

Bibliometric analysis of scientific papers on extracellular vesicles in kidney disease published between 1999 and 2022

“…Camussi Giovanni (corresponding author) from the Department of Medical Sciences, University of Turin, was the top 5 prolific author for EVs in kidney disease. He mainly focused on the association of EVs with kidney-related diseases (AKI, CKD, systemic lupus erythematosus, ANCA-vasculitis, kidney fibrosis, and diabetic nephropathy) ( Biancone and Camussi, 2014 ; Collino et al, 2015 ; Lener et al, 2015 ; Bussolati and Camussi, 2017 ; Dimuccio et al, 2020 ; Grange et al, 2020 ; Kholia et al, 2020 ; Ramírez-Bajo et al, 2020 ; Tetta et al, 2020 ; Kholia et al, 2021 ; Mazzariol et al, 2021 ; Bruno et al, 2022 ; Franzin et al, 2022 ). Additionally, Cortes Raquel (corresponding author) from the Cardiometabolic and Renal Risk Research Group, INCLIVA Biomedical Research Institute, was the top 4 prolific author for EVs in kidney-related diseases.…”

Blood–Brain Barrier Biomarkers before and after Kidney Transplantation

“…Plasma EV preparations were diluted (1:1000) in sterile 0.9% saline, and analyzed with NanoSight LM10 (Nanosight, Amesbury, UK) equipped with Nanoparticle Analysis Systems and NTA 1.4 analytical software to evaluate EV size and concentration. Acquisitions were performed at a camera level setting of 14, and three 30 s videos were recorded for each sample: the number of total EVs (NTA/mL) was obtained by multiplying the value given by the instrument for the dilution for analysis by the ml amount in which EVs were resuspended [ 78 , 79 ].…”

Emerging role of extracellular vesicles in kidney diseases

“…Western blotting can identify exosome markers, such as CD63, CD9, and CD81. However, a single method cannot confirm the identity of EVs, and a combination of nanoparticle tracking analysis, electron microscopy, and western blotting is requires ( Franzin et al, 2022 ).…”