Cell-specific Extracellular Vesicles and Their miRNA Cargo... : Transplantation (original) (raw)

Original Basic Science

Cell-specific Extracellular Vesicles and Their miRNA Cargo Released Into the Organ Preservation Solution During Cold Ischemia Storage as Biomarkers for Liver Transplant Outcomes

Vidal-Correoso, Daniel BSc1; Mateo, Sandra V. BSc1; Muñoz-Morales, Ana M. BSc1; Lucas-Ruiz, Fernando PhD2,3; Jover-Aguilar, Marta BSN1; Alconchel, Felipe PhD, MD1,4; Martínez-Alarcón, Laura PhD1; Sánchez-Redondo, Sara BSc5; Santos, Vanesa BSc5; López-López, Víctor PhD, MD1,4; Ríos-Zambudio, Antonio PhD, MD1,4; Cascales, Pedro PhD, MD1,4; Pons, José Antonio PhD, MD1,6; Ramírez, Pablo PhD, MD1,4; Pelegrín, Pablo PhD1,7; Peinado, Héctor PhD5; Baroja-Mazo, Alberto PhD1

1Molecular Inflammation Group, University Clinical Hospital Virgen de la Arrixaca, Biomedical Research Institute of Murcia (IMIB-Pascual Parrilla), Murcia, Spain.

2Experimental Ophthalmology Group, Biomedical Research Institute of Murcia (IMIB-Pascual Parrilla) & Ophthalmology Department, Universidad de Murcia, Murcia, Spain.

3Institute of Neuroimmunology and Multiple Sclerosis, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

4General Surgery and Abdominal Solid Organ Transplantation Unit, University Clinical Hospital Virgen de la Arrixaca, Murcia, Spain.

5Microenvironment & Metastasis Group, Molecular Oncology Program, Spanish National Cancer Research Centre (CNIO), Madrid, Spain.

6Hepatology and Liver Transplant Unit, University Clinical Hospital Virgen de la Arrixaca, Murcia, Spain.

7Department of Biochemistry and Molecular Biology B and Immunology, Faculty of Medicine, University of Murcia, Murcia, Spain.

Received 19 December 2023. Revision received 24 January 2024.

Accepted 16 February 2024.

S.V.M., A.M.M.-M., and F.L.-R. contributed equally to this work.

All authors read and approved the final version of the article. A.B.-M. participated in the design of the work. A.B.-M., P.P., V.L.-L., F.A., J.A.P., P.R., and H.P. participated in the discussion and review of the article and verified the underlying data. A.B.-M., D.V.-C, S.V.M., A.M.M.-M., F.L.-R., S.S.-R., V.S., and H.P. carried out all the experiments and statistical analysis. A.R.-Z., P.C., P.R., V.L-L., and F.A. organized the intraoperating room collection of samples. M.J.-A. and L.M.-A. collected all the clinical data from patients. JA.P. carried out the follow-up of the patients.

A.B.-M. was funded by Instituto Murciano de Investigación Biosanitaria-Pascual Parrilla (IMIB22/CI/11), Fundación Mutua Madrileña (AP171362019), and Instituto de Salud Carlos III (PI20/00185), co-funded by the European Union. S.V.M. was funded by a predoctoral grant from Instituto de Salud Carlos III (FI21/00073). Funding sources provided financial support but had no involvement in study design, collection, analysis, and interpretation of data.

A.B.-M., P.P., and L.M.-A. are co-founders of Viva In Vitro Diagnostics SL, a company focused on utilizing the NLR Family Pyrin Domain Containing 3 inflammasome as a disease biomarker. A.B.-M. is also co-inventor listed on a provisional patent application for an in vitro method predicting organ transplant rejection. However, it is important to note that this research was conducted independently, without any commercial or financial associations that might be considered a conflict of interest. The other authors declare no conflicts of interest.

Supplemental digital content (SDC) is available for this article. Direct URL citations appear in the printed text, and links to the digital files are provided in the HTML text of this article on the journal’s Web site (www.transplantjournal.com).

Correspondence: Alberto Baroja-Mazo, PhD, Campus de Ciencias de la Salud, Edificio LAIB, Office 4.21, Ctra, Buenavista s/n, 30120 Murcia, Spain. ([email protected]); or José Antonio Pons, PhD, MD, Servicio de Gastroenterología y Hepatología, Hospital Clínico Universitario Virgen de la Arrixaca, Ctra, Cartagena s/n, 30120 Murcia, Spain. ([email protected]).

Abstract

Background.

Liver transplantation (LT) is crucial for end-stage liver disease patients, but organ shortages persist. Donation after circulatory death (DCD) aims to broaden the donor pool but presents challenges. Complications like acute rejection, hepatic artery thrombosis, and biliary issues still impact posttransplant prognosis. Biomarkers, including extracellular vesicles (EVs) and microRNAs (miRNAs), show promise in understanding and monitoring posttransplant events. This study explores the role of EVs and their miRNA cargo in LT, including their potential as diagnostic tools.

Methods.

EVs from intrahepatic end-ischemic organ preservation solution (eiOPS) in 79 donated livers were detected using different techniques (nanosight tracking analysis, transmission electron microscopy, and flow cytometry). EV-derived miRNAs were identified by quantitative real time-polymerase chain reaction. Bioinformatics analysis was performed using the R platform.

Results.

Different-sized and origin-specific EVs were found in eiOPS, with significantly higher concentrations in DCD compared with donation after brain death organs. Additionally, several EV-associated miRNAs, including let-7d-5p, miR-28-5p, miR-200a-3p, miR-200b-3p, miR-200c-3p, and miR-429, were overexpressed in DCD-derived eiOPS. These miRNAs also exhibited differential expression patterns in liver tissue biopsies. Pathway analysis revealed enrichment in signaling pathways involved in extracellular matrix organization and various cellular processes. Moreover, specific EVs and miRNAs correlated with clinical outcomes, including survival and early allograft dysfunction. A predictive model combining biomarkers and clinical variables showed promise in acute rejection detection after LT.

Conclusions.

These findings provide new insights into the use of EVs and miRNAs as biomarkers and their possible influence on posttransplantation outcomes, potentially contributing to improved diagnostic approaches and personalized treatment strategies in LT.