Alzheimer disease (AD) is a neurodegenerative disorder characterized by significant cognitive deficits, behavioral changes, sleep disorders and loss of functional autonomy. The number of patients suffering from AD is growing rapidly as the population ages worldwide. AD represents major cause of dementia in occidental countries. New revised AD diagnosis criteria were proposed. They highlighted the reduction of hippocampic volumetry by MRI, the reduction of glucidic metabolism in specific brain regions as well as alteration of CSF biomarkers: decrease of Aß1-42 peptides, increase of Tau and P-tau proteins. All these biomarkers help the clinical diagnosis with a sensitivity and specificity around 80% but the challenge of AD requires biomarkers beyond these probability and detectable in the blood. The use of news biomarkers in the field of the early diagnosis becomes a requirement because of the current development of therapeutic strategies aiming at slowing down, to even block the neurodegenerative process. Moreover, research in the field of biomarkers leads to relevant data to help elucidate Alzheimer pathophysiological mechanisms. One of the most interesting approaches is the analysis of inflammatory processes involved in AD. Incidentally, recent data suggested that AD pathological processes would produce disease-specific molecular changes in the blood, specifically on secreted signalling proteins, including cytokines, chemokines, and growth factors, as these are the primary means of communication between cells. Genomic investigation in human, as well as in AD animal models, also pointed out the implication of inflammatory molecules such as TNFs in the disease. We propose here further investigations on the involvement of inflammatory processes in AD. This approach is innovative in the field of blood biomarkers as it combines detection and identification of pro-inflammatory factors in human samples with dementia, and in transgenic animal models. A first part our project is based on the targeted and multiplex exploration and validation of inflammatory factors eventually identified in previous proteomic and genomic works. Both microfluidic and biochip multiplex technologies will be utilized. This will be done in patients (AD, prodromal AD, other dementia and inflammatory peripheral and central nervous system) and in a relevant AD transgenic mouse model: THY-Tau22. Data generated will be analysed with state-of-the art bioinformatics. We will then come full circle by cross-validating mouse and human models. In a second phase, putative biomarker candidates will be explored using immuno-histological approaches in mice, and will be measured in the CSF of patients. Our work program will therefore rationalize the identification and characterization of peripheral biomarkers of AD and help decipher the exact function of inflammation in neurodegeneration eventually leading to new therapeutic targets.

The etiologic diversity of TAAD, our ability to have access to human clinical cohorts and DNA collections, to diseased as well as normal human aortic tissues, and to obtain primary cultures of aortic cells, provide us with a unique opportunity to progress further in the pathology of these frequent and highly morbid cardiovascular diseases. The aim of our project is to link functional genomics to pathophysiology of aneurysms and dissections of the human thoracic ascending aorta (TAAD) whatever their aetiology, genetic and non-genetic, through final common pathways involving common risk factors of dissection, epigenetic model of SMC expression regulation, SMC disappearance and involvement of tissue proteolysis. As compared to current studies, mainly focused on monogenic forms(13)and on a possible direct relationship between gene mutations and TAAD phenotype, the originality and novelty of our project is its translational genomic approach from highly morbid pathologies to molecular effectors, possible targets for potential new diagnostic and therapeutic tools. For this purpose we have a number of resources, methods, and know-how at our disposal, including clinical, genetic and tissue/cell databases, experimental models in rat and mice, as well as biological and computational expertise in the analysis of next-generation sequencing and other high-throughput data that will be essential to adapt genomics-scale approaches to the study of vascular pathobiology. Advances expected from this project are to further delineate 1) the relationship between genetic defects and clinical spectrum in humans, of both mutations in newly recognised genes obtained from large TAA families or within known genes in relation to gene modulators. 2) the biological alterations observed in the human TAA from diverse aetiologies including epigenetic alteration observed in the smooth muscle cells. This may transfer into therapeutic target (cf. losartan studies in progress in patients with Marfan syndrome, derived from the TGFB hypothesis generated in mouse). 3) the pathway linking different genetic defects to a similar phenotype (aortic aneurysm) using both human aortic wall coming from TAA from diverse aetiologies and animal models (now is available a mouse model KI for FBN1, i.e. secondary to an anomaly of the extracellular matrix; we propose to create a mouse model KI for MYH11, i.e. secondary to an anomaly in a contractile protein). 4) animal models of aortic dissection

Traditionally, slurry and manure have been used in agriculture as a mean for enriching the soil in nutrients (N, P). Thus, each year in France, 300 million tonnes of animal waste mainly from cattle and pig farms, are spread on the land. Thus, through the spreading of animal waste, hormones and pharmaceutical compounds are dispersed into the soil, representing a source of pollution for both soil and aquatic environment. The increase in the size of livestock farms, along with their localised concentration, has led to problems of nitrate pollution which appear to have been resolved by installing treatment facilities on the big farms. However, such facilities have not been designed for their capacity to eliminate micro-pollutants such as steroid hormones and pharmaceutical compounds. Yet, if waste treatment systems represent a concentrated source of possible contamination of the environment by such as micro-pollutants, by the same token, they are also a point of convergence prior to their eventual dispersal. For this reason, such facilities provide a focal point for intervention aimed at reducing the impact of these molecules on the ecosystems receiving the discharged waste. The aims of this study project are, in the first place: - to investigate the fate of estrogens and antibiotics present in cattle and pig manure during waste storage and treatment; - to determine the transfer of pollutants to the soil and to the plant through waste spreading on land; - to identify the parameters involved in the degradation of the estrogens and antibiotics contained in animal waste; - to optimise the breakdown and elimination of these micro-pollutants; and additionally: - to determine, through the use of luminescent cell lines, the endocrine and toxic effects of pig manure and the matter derived from its treatment; - to determine the impact of antibiotics from animal waste in the development of antibiotic resistance in soil micro-organisms: and - to identify and characterise other estrogenic compounds present in manure. These objectives necessarily involve monitoring the concentrations of estrogens and pharmaceutical compounds in cattle and pig manure storage and treatment units and at each of the different stages in the treatment process (storage pond, reactor tank, sludge…). The transfer of these pollutants to the soil and to the plant, through waste spreading on land, will be analysed in controlled systems. It will enable to determine the real fluxes of pollutants and to quantify their transfer to the soil and plants. In addition to this type of “balance study” there is a need of laboratory investigations to identify the parameters determining the dynamics of the micro-pollutants (adsorption, volatilisation, degradation). Within this framework, two approaches to the elimination of estrogenic and antibiotic compounds will be studied and optimised : biological degradation (aerobic and anaerobic) and heat treatment. An analysis of, in first place, the estrogenic activity and the global toxicity of the manure throughout the treatment process, using bioluminescent cell lines; and additionally, the study of antimicrobial resistance in soils receiving animal waste, will enable us to assess the manure's overall impact for the environment. Finally, the identification of other estrogenic compounds in the animal waste will be carried out using the combined techniques of affinity chromatography and structural chemistry (particularly high resolution mass spectrometry). These studies will lead to an understanding of the eco-dynamics and eco-toxicology of estrogens and pharmaceutical compounds in the different manure storage and treatment systems. The work will also enable the parameters involved in the degradation of such micro-pollutants to be defined, thus resulting in enhanced control of the process and a consequent lessening of their impact on the environment.