Proteome Investigation of Rat Lungs subjected to Ex Vivo Perfusion (EVLP).

Valentina Roffia,Rossana Rossi,Francesco Blasi,Franco Valenza,Caterina Lonati,Marco Mantero,Dario Silvestre,Stefano Gatti,Pierluigi Mauri,Antonella De Palma,Daniele Dondossola

doi:10.3390/molecules23123061

Abstract

Ex vivo lung perfusion (EVLP) is an emerging procedure that allows organ preservation, assessment and reconditioning, increasing the number of marginal donor lungs for transplantation. However, physiological and airflow measurements are unable to unveil the molecular mechanisms responsible of EVLP beneficial effects on lung graft and monitor the proper course of the treatment. Thus, it is urgent to find specific biomarkers that possess these requirements but also accurate and reliable techniques that identify them. The purpose of this study is to give an overview on the potentiality of shotgun proteomic platforms in characterizing the status and the evolution of metabolic pathways during EVLP in order to find new potential EVLP-related biomarkers. A nanoLC-MS/MS system was applied to the proteome analysis of lung tissues from an optimized rat model in three experimental groups: native, pre- and post-EVLP. Technical and biological repeatability were evaluated and, together with clustering analysis, underlined the good quality of data produced. In-house software and bioinformatics tools allowed the label-free extraction of differentially expressed proteins among the three examined conditions and the network visualization of the pathways mainly involved. These promising findings encourage further proteomic investigations of the molecular mechanisms behind EVLP procedure.

Highlights

Ex vivo lung perfusion (EVLP) is an emerging procedure proposed for organ preservation and reconditioning which can increase the number of lungs available for transplantation without compromising its success [1]
IIIM21 (X axis) was compared with that of IIIN23 (Y axis), both belonging to the post-EVLP condition: R2 = 0.946 and y = 0.962. These findings demonstrated the good repeatability of the proteomic approach adopted, because the reduced oscillations of spectral count (SpC) values are a robust starting point for subsequently label-free investigations on protein profiles
The EVLP procedure cannot be solely evaluated on the basis of physiological measurements

Summary

Introduction

Ex vivo lung perfusion (EVLP) is an emerging procedure proposed for organ preservation and reconditioning which can increase the number of lungs available for transplantation without compromising its success [1]. This derives from the fact that organs, previously considered unsuitable for transplantation, thanks to EVLP are safely used and show similar or even better performances than those of lungs transplanted immediately after procurement In these circumstances, EVLP application could represent a benefit for all donor lungs prior to implantation (so that it could be inserted in routine transplant protocols), but the evidence-based medical data collected so far are still insufficient [4] and there are different EVLP systems and application procedures adopted worldwide [5,6,7]. Traditional methods based on reactive medicine (physiological examinations, medical history and airflow measurements) are not sufficient to fully satisfy these requirements

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