Asthma and cardiovascular (CV) diseases are two common conditions with important public health and economic burden. Despite growing evidence that asthma is associated with increased risk of major CV events, the mechanisms by which asthma may affect the risk for CV events remain poorly understood. In particular, whether asthma and CV diseases share common etiological processes (such as anthropometric, lifestyle, social, environmental and/or genetic factors), or whether CV diseases are a consequence of some asthma characteristics (such as asthma treatments or systemic inflammation) remains unknown. The limited knowledge on this mechanism hampers preventive intervention. We aim to disentangle the complex association between asthma and early markers of CV risk, in order to provide new directions in clinical management of patients with asthma and in preventive intervention to prevent CV comorbidities in asthma. To reach this objective, specific aims are: (1) WP1: To collect new data, the 4th follow-up at 30 years of the EGEA (Epidemiological study on the Genetics and Environment of Asthma) cohort to accurately assess: - anthropometric, lifestyle, social and environmental factors through questionnaire, smartphone application, validated 24-h dietary records, passive sampler for individual NO2 assessment, geographical information system-based models; - treatments, hospitalizations, causes of death and long-term illnesses through data linkage with health administrative databases (SNDS); - phenotypic information through detailed questionnaires and clinical examination of the participants including anthropometric measures, lung function, 6-minute walk test, blood pressure and markers of cardiovascular risk (aortic pulse wave velocity [aPWV], Coronary Calcium Score (CAC) assessed by computed tomographic scanner, and biomarkers providing prognostic information on CV risk that will be measured longitudinally (hypersensitive CRP, IL6, hypersensitive Troponine (I), NT-proBNP, and soluble ST2)); (2) WP2: To characterize the longitudinal association of asthma and asthma specific phenotypes with markers of CV risk (aPWV, CAC and biomarkers of CV risk); (3) WP3: To clarify the causal association between asthma and markers of CV risk. Two alternative explanations will be investigated: 1) how much the co-occurrence of asthma and markers of CV risk might be explained by the presence of shared anthropometric, lifestyle, social environmental and genetic common causes; and 2) the role of asthma on markers of CV risk through causal direct and indirect effects (e.g. mediated by asthma treatments or inflammatory pathways). One major asset of the EGEA_30years program is the new follow-up at 30 years of a unique existing cohort including a group of asthma cases recruited in chest clinics, their first-degree relatives and a group of controls (total n=2120), particularly well characterized across the different follow-ups (clinical examination, biomarkers, lifestyle, social, environment, genetic (GWAS), epigenetic (methylome) and metabolomic data) and underpinned by a rich biobank. Given the high follow-up rates achieved in previous EGEA follow-ups, the family and multicentric design of the study, we anticipate that ~1300 individuals will complete the 30-year follow-up questionnaire including ~1000 participants with a clinical examination. Further strengths relate to the expertise of the consortium in setting-up and coordinating cohorts, in asthma and CV research, in analysing large scale omic and exposome data and in applying advanced statistical techniques including cluster and mediation analyses. Our multidisciplinary program will provide tools for identifying and prioritizing determinants of CV risk in asthma and feed into risk prediction, new directions in clinical management as well as development of preventive interventions in asthma. EGEA_30years may unravel actionable levers of life-threatening CV comorbidities in asthma.

The microsporidium Nosema ceranae (a fungi) and the ectoparasite Varroa destructor (a mite) are common biotic stressors of honeybees that produce serious damages to the colonies. The parasitic cycle of V. destructor is composed of a phoretic and reproductive phase. During the latter, the parasite enters brood cells, reproduces and feeds on the hemolymph and fat bodies of bee pupae impinging on their immune system resulting in more frequent physical deformities in newborn bees. In addition, N. ceranae, a gut intracellular parasite, causes nosemosis, the most widespread disease of adult bees. As the main source of lipids and proteins in honey bees, pollen is a major nutrient involved in development and health since many studies have shown its role in shaping honeybee traits, especially immune competence and tolerance against pathogens and parasites. The aim of this project is to investigate the impact of pollen supplementation in adult honeybees exposed to Nosema and previously to Varroa at the larval stage. Impacts of Varroa parasitism in laboratory conditions will be investigated on bee larval growth and on the microbiome in emerging bees. Long term effects on these experimental bees will examine their tolerance and resistance against Nosema exposure both in lab and semi-field conditions. We will focus on the relationships existing between gene expressions, peptidome/proteome modulation and phenotypic traits. We will describe how biotic stressors effects are coupled to interactions between feeding behavior and the microbiota composition. Given our joint expertise, we propose a unique experimental design. This includes impact of nutrition at adult stage and its interaction with most common pathogens, Varroa and Nosema, in both laboratory and semi-field experiments in a simple 4-Tunnels disposition. This interdisciplinary project allows us to examine the effects of those stressors observed all along the beekeeping season.

SCIENTIFIC CONTEXT. Unicellular organisms need to adapt to rapid and unanticipated changes in their environment. Spores ensure their survival by encapsulating the genome in a protective configuration while awaiting optimal growth conditions. Indeed, yeast spores enter into a quiescent state with minimal metabolic activity and are surrounded by a thick protective wall. Yet, protecting the integrity of the genome in these conditions involves not only a dramatic decrease in transcriptional activity, an extreme nuclear compaction, but also the capacity to completely revert these processes to allow germination. The mechanisms involved in the establishment of genomic quiescence in spores, the protection of their genome and its reactivation, remain unknown. OBJECTIVE. Several lines of evidence show that chromatin is highly compacted in spores. In addition, histone H4 is hyperacetylated and this modification is essential for spore viability. This observation seems counter-intuitive because H4 acetylation (H4ac) is usually associated with transcription activation and open chromatin. Therefore, the general objectives of this project are to understand (i) the mechanisms by which H4 is hyperacetylated in spores, (ii) how H4ac is compatible with quiescence in spores and their chromatin compaction, (iii) whether and how H4ac prepares genome reactivation observed during early germination. IMPACT. Through EpiSpores, we will improve our general knowledge on H4ac signalling pathways, chromatin organisation and transcription regulation. Furthermore, yeast spores provide an alternative model system to investigate the molecular mechanisms of quiescence entry, maintenance and exit in all eukaryotic cells. Finally, our previous work on yeast spores has been translated to the treatment of fungal infections, collectively responsible for 1.5 millions of deaths per year. Our future work exploring chromatin signalling pathways in Candida albicans and their functional role in the virulence of this pathogenic yeast will be based on the technological development of this proposal. CONSORTIUM. This project brings together young researchers, by academic standards, with collective and synergetic expertise in yeast biology, genetics, biochemistry, interactomics, high-throughput genetics, super-resolution microscopy and epigenomic approaches.

Photoacoustic imaging (PAI) is a biomedical imaging technique that provides optical contrasts at depth through the generation of acoustic waves with light. Handheld systems can be made for 3D navigation to image the vasculature and its oxygenation. However, these systems are impacted by limited view artifacts that hide some structures and by low contrast-to-noise when using a sparse array. Photoacoustic Fluctuation imaging (PAFI) enables to solve these two issues and we will develop further this technique towards quantitative view-full SO2 imaging. One of the challenges is to correct the spectral coloring effects due to the tissue surrounding the vessels. To this end, we will employ a highly novel multi-modal combination. Beyond these advances concerning PAFI, this technique still suffers from its low temporal resolution. Relying on deep neural networks (DNN) image enhancement abilities, we will improve the frame rate towards the ultimate limit of SO2 images obtained from single shot multispectral images. A main challenge for DNN is to provide quantitative predictions. To address this issue, FULBOX proposes novel experimental approaches to enable real-time full-view imaging of blood oxygenation, which, coupled to state-of-the-art Ultrasound Doppler will enrich the diagnosis for several pathologies.

Background. Thrombocytopenia is defined as an abnormally low blood platelet count, exposing patients at high risks of hemorrhages. It is a common clinical problem in chemotherapy-treated cancer patients with varying incidence, severity and duration. Platelet are produced by megakaryocytes (MK) in the bone marrow (BM). During this process, MKs must detach and transit from the dense cellular BM microenvironment through the sinusoid vessel wall toward the circulation, a process called intravasation. Being a transient process by nature, its exploration in vivo is still challenging and remain poorly understood. Using high-resolution BM imaging, we recently have identified a new in vivo cellular meta-structure, called PodoPZ, composed of an actomyosin-based network of interconnected podosomes which is located in the peripheral zone of MKs. At this strategic location, the PodoPZ is a privileged site of interactions with the microenvironment, to provide trans-endothelium crossing and thereby ensure successful thrombopoiesis, without damaging the integrity of the sinusoidal wall. Central hypothesis. In MegaPod project, we hypothesize that PodoPZ is a key player in the complex adaptation of MK to microenvironmental cues during its journey from the BM to the bloodstream across the endothelial barrier. By regulating MK intravasation and platelet release, it contributes to platelet recovery during homeostasis and after chemotherapy-induced thrombocytopenia (CIT). Objectives and approaches. The MegaPod program is built on a solid consortium with complementary expertise in megakaryocyte physiology, podosome signaling and mechanics, and state of the art imaging technologies. It will also benefit from a collaboration with the university of Michigan for genetic murine models. Combining innovative genetic and super-resolution imaging approaches in mouse models of bone marrow regeneration, we will determine (1) how drug-induced changes in the BM environment affect PodoPZ organization and function in MK. Our preliminary data on a preclinical models of CIT indicate that the PodoPZ adapts its structure to the radical changes of the stressed BM environment. Comparing physiological situation with CIT mice models, we will provide an overall picture of the dynamic PodoPZ adaptation to the surrounding BM microenvironment, and we will identify specific sets of properties associated to drug-induced thrombocytopenia. (2) how the PodoPZ drives the passage of the MK through the endothelium. We will examine how MK intravasation rely to the coupling between force generation, adhesive or degradative functions of the PodoPZ. (3) what are the specific signaling pathways in the MK supporting the functional regulation of the PodoPZ. We will characterize a new in vitro 3D model mimicking the confined environment of BM in order to control, maintain and manipulate the dynamics of PodoPZ. This in vitro approach will be used as a platform for omics and biochemical approaches necessary to identify new molecular PodoPZ regulators. Relevance and strategy. At the fundamental level, MegaPod project aims to advance our knowledge on the poorly characterized megakaryocyte intravasation in vivo, a process which is absolutely mandatory for efficient platelet release. Our findings are also expected to have an impact for a wider scientific community, as they will also bring new perspectives on how podosomes might function in vivo in other tissues. Indeed, different cell types, including macrophages and osteoclasts, elaborate interconnected pododomes, similar to PodoPZ,and, to date, only a few data have been published on podosome in vivo. At the clinical level, a novel application could stem from this work, whereby targeting megakaryocyte intravasation to ensuring platelet release, rather than stimulating megakaryocyte differentiation, would offer an alternative treatment method for better management of chemotherapy-induced thrombocytopenia in the future.