Use of mass spectrometry imaging and its optimization during the drug development process

La recherche et développement (R&D) au sein de l’industrie pharmaceutique est une étape cruciale pour la découverte de nouveaux médicaments ou biomarqueurs. Le développement de nouveaux traitements innovants est un moteur essentiel du progrès dans la prise en charge de nombreuses maladies, comme la parodontite, ou encore en immuno-oncologie. Ce travail de thèse s’intéresse dans un premier temps à la place de l’imagerie par spectrométrie de masse utilisant une source désorption laser assistée par matrice (MALDI-MSI) lors du développement de nouveaux médicaments, suivie de ses évolutions passées en revue (matrices, instruments et logiciels), avant d’illustrer plusieurs développements réalisés avec cette technologie pour améliorer la sensibilité de détection de composés d’intérêt (médicaments/biomarqueurs), mais aussi la qualité des résultats (qualité des échantillons ou qualité de l’analyse) en vue d’une standardisation. Nos résultats montrent que la mise en place d’une stratégie de dérivation (pour une drogue ou des biomarqueurs), ainsi que l’optimisation des étapes de préparation des échantillons (stabilisation des métabolites, optimisation du dépôt automatisé de matrice) permettent d’améliorer la sensibilité de détection au sein des échantillons. Aussi, la mise en place de contrôles qualité et la validation de méthode de quantification permet d’améliorer la qualité des résultats. Ces développements permettent alors d’aider les industries pharmaceutiques dans les étapes de R&D, en leur permettant de combiner cette technologie à leur arsenal, dans le but de gagner du temps et de l’argent lors des étapes de développement de nouveaux candidats médicaments.Research and development (R&D) in the pharmaceutical industry is a crucial step for the discovery of new drugs or biomarkers. The development of new innovative treatments is a key driver of progress in the management of many diseases, such as periodontitis or immuno-oncology. This thesis work is initially interested in the place of mass spectrometry imaging using a matrix-assisted laser desorption ionization source (MALDI-MSI) during the development of new drugs, followed by its evolution reviewed (matrices, instruments and software) before illustrating several developments made with this technology to improve the detection of the compounds of interest (drugs / biomarkers), but also the quality of the results (quality of the samples or the quality of the analyze) with a view to standardization. Also, the implementation of quality controls and the validation of the quantification method improves the quality of the results. These developments then help pharmaceutical industries in the R&D stages, allowing them to combine this technology with their arsenal, in order to save time and money during the development stages of new drug candidates

Mechanisms of innate events during skin reaction following intradermal injection of seasonal influenza vaccine

International audienceThe skin plays a crucial role in host defences against microbial attack and the innate cells must provide the immune system with sufficient information to organize these defences. This unique feature makes the skin a promising site for vaccine administration. Although cellular innate immune events during vaccination have been widely studied, initial events remain poorly understood. Our aim is to determine molecular biomarkers of skin innate reaction after intradermal (i.d.) immunization. Using an ex vivo human explant model from healthy donors, we investigated by NanoLC-MS/MS analysis and MALDI-MSI imaging, to detect innate molecular events (lipids, metabolites, proteins) few hours after i.d. administration of seasonal trivalent influenza vaccine (TIV). This multimodel approach allowed to identify early molecules differentially expressed in dermal and epidermal layers at 4 and 18 h after TIV immunization compared with control PBS. In the dermis, the most relevant network of proteins upregulated were related to cell-to-cell signalling and cell trafficking. The molecular signatures detected were associated with chemokines such as CXCL8, a chemoattractant of neutrophils. In the epidermis, the most relevant networks were associated with activation of antigen-presenting cells and related to CXCL10. Our study proposes a novel step-forward approach to identify biomarkers of skin innate reaction. SIGNIFICANCE: To our knowledge, there is no study analyzing innate molecular reaction to vaccines at the site of skin immunization. What is known on skin reaction is based on macroscopic (erythema, redness…), microscopic (epidermal and dermal tissues) and cellular events (inflammatory cell infiltrate). Therefore, we propose a multimodal approach to analyze molecular events at the site of vaccine injection on skin tissue. We identified early molecular networks involved biological functions such cell migration, cell-to-cell interaction and antigen presentation, validated by chemokine expression, in the epidermis and dermis, then could be used as early indicator of success in immunization

Caffeine intake exerts dual genome-wide effects on hippocampal metabolism and learning-dependent transcription

Caffeine is the most consumed psychoactive substance worldwide. Strikingly, molecular pathways engaged by its regular consumption remain unclear. We herein addressed the mechanisms associated with habitual (chronic) caffeine consumption in the mouse hippocampus using untargeted orthogonal-omics techniques. Our results revealed that chronic caffeine exerts concerted pleiotropic effects in the hippocampus, at the epigenomic, proteomic and metabolomic levels. Caffeine lowers metabolic-related processes in the bulk tissue, while it induces neuronal-specific epigenetic changes at synaptic transmission/plasticity-related genes and increased experience-driven transcriptional activity. Altogether, these findings suggest that regular caffeine intake improves the signal-to-noise ratio during information encoding, in part through a fine-tuning of metabolic genes while boosting the salience of information processing during learning in neuronal circuits.This work was supported by grants from Hauts-de-France (PARTEN-AIRR, COGNADORA; START-AIRR, INS-SPECT) and Programs d’Investissements d’Avenir LabEx (excellence laboratory) DISTALZ (Development of Innovative Strategies for a Transdisciplinary approach to ALZheimer’s disease) and EGID (European Genomic Institute for Diabetes ANR-10LABX-46). Our laboratories are also supported by ANR (GRAND to LB, ADORATAU, ADORASTrAU, METABOTAU to DB and BETAPLASTICITY to JSA), COEN (5008), Fondation pour la Recherche Médicale, France Alzheimer/Fondation de France, FHU VasCog research network (Lille, France), Fondation Vaincre Alzheimer (ADOMEMOTAU), European Foundation for the Study of Diabetes (EFSD to JSA), Fondation Plan Alzheimer as well as Inserm, CNRS, Université Lille, Lille Métropole Communauté Urbaine, DN2M. KC hold a doctoral grant from Lille University. VG-M was supported by Fondation pour la Recherche Médicale (SPF20160936000). CM was supported by Région Hauts753 30 754 de-France. ALB is supported by CNRS, Unistra (Strasbourg, France), ANR-16-CE92-0031 755 756 757 758 759 760 761 762 (EPIFUS), ANR-18-CE16-0008-02 (ADORASTrAU), Alsace Alzheimer 67, France Alzheimer (AAP SM 2017 #1664). IP is supported by Fondation pour la Recherche Médicale (SPF201909009162). CEM is grateful for the support by the Alzheimer Forschung Initiative e.V. (AFI, Düsseldorf, Germany). LC was funded by SIF Italian Society of Pharmacology. RAC was supported by LaCaixa Foundation (LCF/PR/HP17/52190001) and FCT (POCI-01-0145-FEDER-03127). Santa Casa da Misericórdia (MB-7-2018) and CEECIND/01497/2017 to LVL