High-multiplexed monitoring of protein biomarkers in the sentinel Gammarus fossarum by targeted scout-MRM assay, a new vision for ecotoxicoproteomics

Julien Faugere (1) , Duarte Gouveia (2, 3) , Sophie Ayciriex (1) , Arnaud Chaumot (3) , Christine Almunia (2) , Adeline François (3) , J. Armengaud (2) , Jérôme Lemoine (1) , Olivier Geffard (3) , Davide Degli Esposti (3) , Arnaud Salvador (1)

1 ISA-ANABIO-MS - Analyse biomoléculaire par spectrométrie de masse - Biological Analysis by Mass Spectrometry
2 LI2D - Laboratoire Innovations technologiques pour la Détection et le Diagnostic
3 RiverLy - RiverLy - Fonctionnement des hydrosystèmes

Ecotoxicoproteomics employs mass spectrometry-based approaches centered on proteins of sentinel organisms to assess for instance, chemical toxicity in fresh water. In this study, we combined proteogenomics experiments and a novel targeted proteomics approach free from retention time scheduling called Scout-MRM. This methodology will enable the measurement of simultaneously changes in the relative abundance of multiple proteins involved in key physiological processes and potentially impacted by contaminants in the freshwater sentinel Gammarus fossarum. The development and validation of the assay were performed to target 157 protein biomarkers of this non-model organism. We carefully chose and validated the transitions to monitor using conventional parameters (linearity, repeatability, LOD, LOQ). Finally, the potential of the methodology is illustrated by measuring 277peptide-plex assay (831 transitions) in sentinel animals exposed in natura to different agricultural sites potentially exposed to pesticide contamination. Multivariate data analyses highlighted the modulation of several key proteins involved in feeding and molting. This multiplex-targeted proteomics assay paves the way for the discovery and the use of a large panel of novel protein biomarkers in emergent ecotoxicological models for environmental monitoring in the future.Biological significance: The study contributed to the development of Scout-MRM for the high-throughput quantitation of a large panel of proteins in the Gammarus fossarum freshwater sentinel. Increasing the number of markers in ecotoxicoproteomics is of most interest to assess the impact of pollutants in freshwater organisms. The development and validation of the assay enabled the monitoring of a large panel of reporter peptides of exposed gammarids. To illustrate the applicability of the methodology, animals from different agricultural sites were analysed. The application of the assay highlighted the modulation of some biomarker proteins involved in key physiological pathways, such as molting, feeding and general stress response. Increasing multiplexing capabilities and field test will provide the development of diagnostic protein biomarkers for emergent ecotoxicological models in future environmental biomonitoring programs.

Mots clés: Mass spectrometry; Protein quanti fication; Scout-MRM; Ecotoxicology; Biomarker; Multiplex approach

Domaines: Chimie analytique; Chimie; Sciences de l'environnement

Julien Faugere, Duarte Gouveia, Sophie Ayciriex, Arnaud Chaumot, Christine Almunia, et al.. High-multiplexed monitoring of protein biomarkers in the sentinel Gammarus fossarum by targeted scout-MRM assay, a new vision for ecotoxicoproteomics. Journal of Proteomics, 2020, 226, ⟨10.1016/j.jprot.2020.103901⟩. ⟨hal-02903090⟩