We present a methodological development for glycomics - a field which lags far behind its counterparts genomics and proteims in terms of available analytical tools. The proposed method will focus on profiling glycans epitopes (acetylated sialic acid and sulfatated extremities) which is currently beyond the state of the art of analytical chemistry and impairs the comprehensive characterisation of glycosylation profiles of proteins. The original concept underlying the proposition is the use of IR ion spectroscopy coupled with three dimension of separation (LC, IMS and MS) to resolve all existing isomeric patterns present in a heterogeneous mixture. The project brings together an interdisciplinary consortium of acknowledged experts in the fields of glycomics, ion spectroscopy, glycan synthesis and immunology.?To demonstrate the future impact of the method for glycosciences, we propose a case study: the early diagnostic of rheumatoid arthritis.

Survivors of sepsis – a severe infection leading to intensive care unit (ICU), are more and more numerous because of greater hospital care and an increasing aged population. However, their stay in Intensive Care Unit (ICU) is responsible for sustained consequences, known as the post-intensive care syndrome, that accelerate physiological aging and alter long-term prognosis. As such, survivors develop muscle weakness during their hospital stay and it persists months after ICU discharge. As in physiological aging, mitochondrial dysfunction and inflammation are key mechanisms in long-lasting ICU-acquired weakness. Importantly, data from our team and others suggest that inhibition of the Receptor for Advanced Glycation End-products (RAGE) – a multiligand pattern recognition receptor – is of potential interest to reduce those mechanisms. Therefore, we aim to study the effects of RAGE inhibition on sepsis-induced muscle weakness and to understand the underlying mechanisms. However, several obstacles must be overcome: (i) develop a clinically relevant model of sepsis-inducing muscle weakness, (ii) develop specific inhibitors against RAGE, and (iii) set up a device to test these molecules on human muscle. To do so, we will use cellular (differentiated murine and human myoblasts), animal (sepsis induced in mice by injection of heterologous stool), and human (human muscle development by tissue engineering from patient biopsies) models of sepsis-induced muscle weakness. Modulation of RAGE will be achieved by genetic invalidation or synthesis of pharmacological inhibitors of RAGE developed within the consortium. We expect to show that RAGE inhibition improves mitochondrial parameters, reduces inflammation and senescence, attenuates RAGE-related signaling pathways, and restores overall skeletal muscle function. By developing the first tissue bioengineered model of post-septic muscle weakness derived from critical care patients, we will advance our novel patentable inhibitors into the clinic. We hope to make a step towards personalized medicine for weakness acquired after ICU hospitalization. This study is critical to understanding the pathophysiology of sepsis-accelerated muscle aging, and to proposing a new individualized treatment for a key feature of the post-resuscitation syndrome. In the long run, we hope to improve the patient's quality of life and long-term survival.

Heart failure (HF) affects 2% of the EU population and is an unmet clinical need due to the limited treatment options. HF is associated with alterations of the immune system. Non-Alcoholic Fatty Liver Disease (NAFLD), an emerging health problem associated with metabolic diseases, such as obesity and type 2 diabetes (T2D), is an independent cardiovascular disease risk factor. When NAFLD progresses from simple steatosis to Non-Alcoholic SteatoHepatitis (NASH) with fibrosis, the risk to develop hepatic and cardiac complications increases. However, the mechanisms linking NASH and HF are not understood. Strikingly, monocytes (Mono) and macrophages (Macro) are important contributors to cardiovascular disease and these cells also plays a key role in NASH. Moreover, our preliminary results identify these immune cells as potential link between NAFLD and cardiac dysfunction. We thus hypothesize that NASH and its metabolic alterations impact on the immune compartment, which in turn functionally contributes to inappropriate cardiac remodelling (CR), leading to HF. We propose an ambitious project combining systems biology approaches, state-of-the-art molecular technologies, original preclinical models and human translational studies to: 1. Define the role of NASH in (the aggravation of) CR in mice; 2. Characterize the immune cell alterations in NASH-induced CR with a focus on Mono and Macro; 3. Determine the functional role of thus identified immune cells in CR; 4. Translate our findings to the human pathology by analyzing the impact of NAFLD on cardiac complications (atrial fibrillation and HF) in patients undergoing aortic valve stenosis surgery. The unique combination of expertise in immunology, metabolism and cardiology within our consortium is critical to decipher how immune system alterations link liver pathology to heart disease. Our results will improve our understanding of HF pathophysiology allowing the identification of novel patient management approaches.

The environmental impact of packaging has become a major concern for the food industries, packaging, safety agencies but also consumers. This circular economy will force manufacturers to lighten packaging, recycle and/or reuse it, which implies having the same requirements in terms of health safety as for virgin materials. Indeed, materials in contact with foodstuffs (FCM) can transfer constituents to food by migration. In addition to substances of known origin, FCMs can also contain Non-Intentional Substances (NIS) (impurities, decomposition or reaction products, contaminants resulting from recycling, etc.), often of unknown and unpredictable origin. Most SNIs are neither identified nor quantified, and their toxicity has not been studied. This migration can pose a risk to human health, it must be measured and controlled. European Regulation 10/2011 on plastic materials also requires a risk assessment of SNIs, but to date there is no specific guideline or scientific consensus, making it difficult to assess and manage their risks. In addition, SNIs can be present in all packaging; recycled paper-cardboard, coatings, these can release more substances than their virgin equivalent. The evaluation of MCDA is currently only based on a study of the genotoxicity and the systemic effects of the starting substances, not taking into account endocrine disruption, nor the "cocktail effects" at low dose. Regarding SNI, it is recognized that the traditional approach based on the identification and quantification of all substances followed by their full toxicological characterization is not feasible (in terms of costs, time, quantity available, etc.). A relevant approach consists in using biotests in addition to analytical and physicochemical techniques on all of the substances which migrate. Biotests are already used with mixtures. However, they need to be better characterized in terms of sensitivity / specificity and robustness with complex extracts of MCDA. The first stage of this project will consist in selecting the most sensitive and specific biotests to identify a hazard (genotoxicity or endocrine disruptor) in packaging extracts by applying the “spiking” methodology, which consists of adding reference substances ( positive and negative) in order to verify the expected response. The extract will be split if the answer is a false negative or a false positive, in order to identify the responsible fraction containing the substances causing the unexpected effect. The second step will consist of testing the selected biotests using extracts from finished packaging that have been subjected to particularly SNI-generating processes (including recycled materials) to assess the risk. The innovative nature of this project is to use in parallel, chemical signatures of MCDA extracts and robust biotests in order to generate a database allowing decision-making and packaging security at different stages of their cycle of production life. In addition, it will generate data on the toxicity of new SNIs as well as potential "mixing" effects of the extract. Resulting from a multidisciplinary scientific approach, this project will help packaging manufacturers and their customers (processors, food industries) (1) to better guarantee the safety and conformity of their materials and (2) to encourage their innovation and / or their competitiveness, by offering them relevant and reliable scientific tools.

Clostridioides difficile, an anaerobic Gram-positive spore-forming bacterium, is responsible for a wide spectrum of infections ranging from diarrhea to life-threatening pseudomembranous colitis. The use of antibiotic therapy raises concerns about the selection of antibiotic-resistant bacteria at hospitals. New therapeutic targets should be investigated as alternatives to antibiotic treatments. Polysaccharides (PS) biosynthesis enzymes are encoded in a PS locus where most genes are essential for bacterial viability. We propose therefore the enzymes involved in PS biosynthesis as new therapeutic targets. Moreover, vaccines currently under development target the toxins and may not prevent C. difficile colonization and dissemination. We also propose in this project to evaluate PSII and/or LTA as vaccine component(s) of a vaccine that may prevent C. difficile colonization and dissemination. To that aim, the project will (i) define if either one or both PSII and LTA are essential for bacterial viability, ii) identify specific enzymes involved in PSII or LTA biosynthesis, (iii) target them with inhibitors and (iv) test PSII and LTA as vaccine candidates using an innovative approach. We recently developed a new genetic conditional lethal mutant system and have already obtained antibodies directed against PSII. Using these tools, we showed that the PSII seems to be essential for bacterial viability. The project will be divided into three tasks. The first will determine whether PSII, its anchoring and/or LTA are essential and define at least two enzymes as new therapeutic targets, one of each involved in each PS biosynthesis. The second task will look for inhibitors able to target these specific enzymes, using in silico models and a chemistry approach. The last task will test PSII and LTA as vaccine candidates. This project aims to combat C. difficile infections and prevent them using vaccination.