Malaria is still a major public health problem in 2018, since this infection due to the Plasmodium parasite, present in 91 countries, kills more than 440,000 people every year, with a majority of African children. As observed in most of infectious diseases, the emergence of resistant strains of P. falciparum toward antimalarials is nowadays responsible for a major concern, especially regarding the increasing resistant rate observed toward artemisinin derivatives (ref. treatments for P. falciparum malaria) in Asia, and the subsequent development of multi-resistant strains that could spread worldwide without any therapeutic option. Then, to bypass parasitic resistance, new antimalarials are expected to be active on artemisinin-resistant strains and to possess a novel mechanism of action. It is also crucial to develop new molecules targeting both the asexual (hepatic and erythrocytic) and sexual (gametocytes) stages of the parasite, in a view to block malaria transmission. We previously identified a multi-stage acting lead compound in the thienopyrimidinone series, called Gamhepathiopine (or M1), which fulfills these criteria by acting on erythrocytic, hepatic and sexual stages of P. falciparum. This original molecule displays excellent in vitro activities even on the artemisinin-resistant parasites. In addition, our lead molecule does not exert its antiplasmodial activity by a mechanism of action already described for marketed antimalarials. Such promising results are however tempered by a rapid hepatic metabolization and a poor aqueous solubility, which limit gamhepathiopine activity in vivo. In this context, we thus propose to pharmacomodulate gamhepathiopine to optimize its physico-chemical and pharmacokinetic properties and to potentiate its in vivo activity. To this aim, the metabolic stability issue will be addressed by modifying the two identified sites of metabolization of gamhepathiopine. In addition, the aqueous solubility will be improved by introduction of polar groups and/or synthesis of hydrophilic prodrugs. Additional pharmacomodulation work is also proposed in order to identify new antiplasmodial leads and to modulate the purine-analog character of the scaffold, in the hypothesis of a mode of action related to plasmodial kinase inhibition. The new molecules will be evaluated in vitro on the erythrocytic stages (sensible and resistant strains) and for their cytotoxicity. The most active compounds will be studied 1) for their mutagenicity 2) in vitro against hepatic stages ( (P. yoelii, P. falciparum and P. vivax and for their action on gametocytes and their capacity to block parasite transmission to Anopheles (vector of malaria)) then 3) in vivo, after potential preparation of lipidic nanoemulsions, on a murine model either infected by P. yoelii or humanized and infected by P. falciparum or P. vivax to validate the benefit of the chemical modifications toward PK properties and guarantee the preservation of the multi-stage acting properties. The last part of the project concerns the identification of the gamhepathiopine plasmodial target, in a view to elucidate its novel mechanism of action. A first hypothesis based on M1 chemical structure (purine analogue) and on recent literature data consists in postulating the possible involvement of a plasmodial kinase to explain its antiplasmodial activity. A phospho-proteomic study will thus be conducted to answer this question. In parallel and to extent the study of the mechanism of action of the lead molecule to potential non-kinase targets, an affinity chromatography procedure will be applied to Gamhepathiopine (immobilized on a solid support via a spacer) to try to isolate its target from a plasmodial lysate and to proceed to its identification by MALDI-TOF.

HYPOTHESIS Multiplexed analysis of the diversity and targets of fungal exposome-induced sensitization enables risk modeling and long-term prediction, at the population and individual levels, of lung function, a correlate of longevity and quality of life. OBJECTIVES Main objective: To establish a biomarker pattern of multiplex sensitization to the environmental fungal exposome and allergic inflammation associated with lung function temporal trajectories in asthmatic and healthy adults from the French part of the ECRHS cohort (discovery cohort). The selected biomarkers will address allergic sensitization as revealed by specific antifungal antibodies of isotype E, hereafter denoted as immunoglobulin (Ig) E, using a custom multiplex allergen platform featuring all currently known allergenic fungal genera, and allergic inflammation based on eosinophil activation-related molecules. FUNGIPLEX involves and is coordinated by the French ECRHS clinical and scientific PIs: Pr P. Demoly (FUNGIPLEX coordinator and ECRHS clinical PI, Montpellier), Dr I. Pin (ECRHS clinical PI, Grenoble), Dr C. Neukirch (ECRHS clinical PI, Paris), Pr C. Raherison (ECRHS clinical PI, Bordeaux), Dr B. Leynaert (scientific coordinator of ECRHS France, Paris). FUNGIPLEX study design addressing the endpoint of allergic sensitization to environmental allergens is in compliance with ECRHS ethical and regulatory requirements. Secondary objectives: 1. To test the discovery cohort results in two distinct replication cohorts: a. Pediatric replication of adult findings: Replication in pediatric asthma. This replication arm will test the influence of age, assuming climate and genetic factors are comparable to ECRHS France. It will also add pathophysiological insight into the natural history of fungal sensitization during childhood. It will be performed with the French SAMP cohort comprising 424 preschool and school patients. Pr J. Just (Paris), the clinical PI of SAMP, is involved in FUNGIPLEX as scientific team leader. b. Swedish ECRHS replication of French ECRHS findings. This replication arm will test the influence of climate and genetic variation, assuming demographic and lung function characteristics are comparable to ECRHS France. ECRHS Sweden was designed and conducted similarly to ECRHS France. ECRHS Uppsala comprises 302 patients assessed at the 3 time points. FUNGIPLEX involves the clinical PIs of ECRHS Uppsala: Pr C. Janson and Pr A. Malinovschi. 2. To improve detection of susceptibility to long-term lung and health outcomes at the individual level: FUNGIPLEX will bridge mold exposure questionnaires with corresponding multiplex mold immune sensitization. ORIGINALITY A translational hub from epidemiological data to personalized medicine in the context of fungal exposome. EXPECTED RESULTS In terms of new knowledge, FUNGIPLEX will provide the description of the evolution of the fungal sensitization in relation to clinical outcomes at various times of life and taking different regions into account at the general population level. Of note, some of the fungi have never been considered in population studies. Determinants of this evolution will be investigated taking an EnvWAS approach into account. This information will be used for personal prevention. Clinically, FUNGIPLEX will provide an extensive fungal multiplex assay to be used in other cohorts (COBRA, EGEA, etc.) as well as in clinical routine. FINAL PRODUCT DEVELOPED The multiplex assay developed in the FUNGIPLEX study will help identify the most informative fungal allergens and serve as a basis for tailored multiplex tools. Such tools are needed for the extended or focused investigation of other fungal-related diseases involving IgE, IgG, or IgA in serum or local samples (nasal fluid, sputum), e. g. hypersensitivity pneumonitis, allergic bronchopulmonary mycoses and allergic fungal rhinosinusitis.

Analyses of the health crises which have occurred in recent decades showed that they were also social, economic and political crises which were notably fed by uncertainties about actual severity of the risk, health consequences to be feared, duration of the crisis, or reactions of the population. Public reactions are indeed a major unknown component in crisis management, as such management generally relies on measures requiring active support from the population. Public support also becomes crucial when the crisis extends over time, especially if coercive measures have been implemented. In the current French context of confinement since March 17 due to the COVID-19 global health crisis, we aim to analyze the reactions of the French population to coercive governmental decisions, especially as regards living conditions and health, daily activities including information research, risk perceptions, knowledge and beliefs relating to the disease, opinions and attitudes towards epidemic, heath crisis management and the actors involved. Issues relating to social differentiation and temporal dynamics will be explored in-depth (as public reactions are expected to evolve due to both the epidemic and political measures). Our project provides for a continuous, flexible and reactive tool. Since it is currently impossible to collect face-to-face data or even telephone data (telephone survey platforms are closed due to COVID-19), an online data collection is favored. Using a “rolling poll” technique will then allow a virtually continuous data collection (4 waves of 500 surveyed people each week). For each survey wave, the planned questionnaire (about 20-30 minutes to be filled out) will include a core questionnaire part to allow longitudinal monitoring of knowledge, beliefs and attitudes. Once the actual conditions of confinement being specified, health states of confined individuals (including stress and anxiety) will be assessed as well as their reactions to containment. In addition to the core questionnaire, a variable part (modified according to the waves) aims to account for the evolution of the health situation and the emergence of new issues. In all, our approach therefore aims at “sticking to” the COVID19 crisis as closely as possible by means of a tight supervision by a multidisciplinary Scientific Steering Committee (involving sociology, social psychology, economics, medicine and public health) whose role is first to adapt the questionnaire depending on the evolution of French epidemic situation and major issues. Our project combines both scientific and operational objectives. In the short term, we aim to provide the authorities with a decision-making tool enabling them to monitor real time reactions and to anticipate, at least in part, reactions to a possible “second wave” of the COVID19 epidemic. In addition, feedback on this tool will be provided in the perspective of preparing public responses to future similar health crises. On a scientific ground, better knowledge and understanding of individual reactions to coercive measures is also a major scientific challenge for the researchers involved in this project, notably in view of the social heterogeneity of these reactions and their evolution over time. The collected data could also be used as a main input for implementation of epidemiological models attempting to predict the spread of the epidemic over time and space.