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Evs from Patients with Amr Induce Tubular Senescence and Endmt Through Horizontal Transfer of Mirnas

R. Franzin1, A. Stasi1, F. Sallustio1, S. Bruno2, G. Merlotti3, M. Quaglia3, G. Grandaliano4, P. Pontrelli1, J. Thurman5, G. Camussi2, G. Stallone6, V. Cantaluppi3, L. Gesualdo1, G. Castellano7

1DETO, University of Bari, Bari, Italy, 2University of Turin, Turin, Italy, 3University of Piemonte Orientale, Novara, Italy, 4Università Cattolica Sacro Cuore, Rome, Italy, 5University of Colorado School of Medicine, Aurora, CO, 6University of Foggia, Foggia, Italy, 7University of Milan, Milan, Italy

Meeting: 2022 American Transplant Congress

Abstract number: 1284

Keywords: Graft function, Inflammation, Kidney transplantation, Rejection

Topic: Basic Science » Basic Science » 12 - Immunosuppression & Tolerance: Preclinical & Translational Studies

Session Information

Session Name: Immunosuppression & Tolerance: Preclinical & Translational Studies

Session Type: Poster Abstract

Date: Monday, June 6, 2022

Session Time: 7:00pm-8:00pm

 Presentation Time: 7:00pm-8:00pm

Location: Hynes Halls C & D

*Purpose: Extracellular vesicles (EV) are emerging mediators in several diseases; however, their role in antibody-mediated allograft rejection (AMR) has been poorly investigated.

*Methods: Plasma EVs from 10 Acute and Chronic AMR patients were purified by ultracentrifugation and characterized by Nanoparticle Tracking Analysis. RPTEC and Eahy926 culture were incubated with EVs; EndMT was performed by FACS. Cellular senescence was investigated by qPCR and IHC. mRNA level of C3 and CFH were also measured. Renal biopsies were analyzed for inflammaging (p16INK4a) and EndMT markers (CD31/αSMA) by IHC and IF.

*Results: Renal AMR biopsies showed significant tubular senescence as indicated by p16 expression; p16 was significantly upregulated in Chronic compared with Acute AMR biopsies (p<0.05). The AMR biopsies showed EndMT, as indicated by CD31 decrease and interstitial αSMA upregulation (p<0.05). In AMR patients, EVs appeared to be significantly increased in acute AMR compared to healthy serum controls, in a higher manner than chronic AMR (NTA, p=0.0154). In vitro, the exposure of RTEC to AMR-derived EVs induced senescence as observed by significant increase in SA-β-gal+ cells and p21, with p53 and CYP1B1 increased gene expression (p<0.05). Furthermore, AMR derived EVs induced downregulation in Klotho gene expression in RPTEC. In accordance, AMR derived EVs induced EndMT as observed by FACS with downregulation of CD31, VE-cadherin (CD144) and increase of Vimentin and Collagen I (p=0.025). Finally, EVs from AMR patients induced a significant increase in C3 gene expression with downregulation in CFH in RPTEC. By microarray analysis of EV, miR-604, miR-515-3p, miR-let-7d-5p and miR-590-3p were significantly up-regulated in AMR group, whereas miR-24-3p and miR-29a-3p were downregulated.Conclusions: These results suggest a putative role for circulating AMR derived-EVs in inducing tubular inflammaging and EndMT by mediating complement activation. The EVs cargo characterization might highlight novel targets for new therapeutic intervention.

*Conclusions: These results suggest a putative role for circulating AMR derived-EVs in inducing tubular inflammaging and EndMT by mediating complement activation. The EVs cargo characterization might highlight novel targets for new therapeutic intervention.

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To cite this abstract in AMA style:

Franzin R, Stasi A, Sallustio F, Bruno S, Merlotti G, Quaglia M, Grandaliano G, Pontrelli P, Thurman J, Camussi G, Stallone G, Cantaluppi V, Gesualdo L, Castellano G. Evs from Patients with Amr Induce Tubular Senescence and Endmt Through Horizontal Transfer of Mirnas [abstract]. Am J Transplant. 2022; 22 (suppl 3). https://atcmeetingabstracts.com/abstract/evs-from-patients-with-amr-induce-tubular-senescence-and-endmt-through-horizontal-transfer-of-mirnas/. Accessed May 30, 2025.

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