Title Promoter Affiliations Abstract "Using molecular proximity to refine synapse quantification in neuronal cultures." "Alfonso Gerardo Garcia" "Laboratory of cell biology and histology" "Synapses are specialized connections between neuronal cells that determine the wiringpatterns, which are essential for memory and cognition. Synaptic dysfunction is a commonpathological hallmark of neurodevelopmental and - degenerative conditions. Thus, accurateand reliable quantification of synaptic state and number in neuronal networks is crucial. Inprevious work, we have shown that primary neuronal cultures from rodents preserve manymorphological and functional properties of in vivo neuronal networks and can be used toevaluate the impact of chemo-genetic perturbations on synaptic state. However, the largenumber of synapses on the one hand, and the variable specificity of existing microscopytechniques on the other hand, makes synapse quantification in these cultures a balancing actbetween accuracy and throughput. To improve both, we propose to make use of a novelmethod, termed Proximity Ligation Assay (PLA), that can directly visualize molecularinteractions using a standard fluorescence microscope. To do so, we will first identify anoptimal set of trans-synaptic protein interactors. Then, we will validate the PLA technique anduse it to measure synapse density after application of targeted perturbations. By applying PLAto trans-synaptic proteins, we aim at detecting true synapses with superior specificity. Thisshould enhance the sensitivity with which we can detect changes in synapse density, andtherefore it has the potential to accelerate the identification of synaptic modulators in ourongoing screening efforts." "Exploring the anti-inflammatory potential of Rothia mucilaginosa in chronic lung diseases." "Paul Cos" "Ghent University, Laboratory for Microbiology, Parasitology and Hygiene (LMPH)" "In this project, we will confirm the anti-inflammatory effects of Rothia mucilaginosa in complex and physiologically relevant in vitro models of CF and COPD lung inflammation, as well as in in vivo mouse models. The anti-inflammatory compound(s) produced by this bacterium will be identified and the mode of action unveiled." "The role of autophagy in the prevention of oxidative stress and cardiovascular disease by olive polyphenols." "Nina Hermans" "Physiopharmacology (PHYSPHAR), Natural Products and Food - Research and Analysis (NatuRA)" "Atherosclerotic plaque rupture is the leading cause of acute cardiovascular syndromes and is responsible for 3.9 million deaths in Europe every year. Preventive strategies are greatly needed to reduce the health care burden of cardiovascular disease (CVD). It is well-known that the Mediterranean diet results in a lower CVD risk, with virgin olive oil as an important element. Many of the health-promoting effects are ascribed to the olive polyphenols. Over the years, studies have shown that olive polyphenols reduce oxidative stress and inflammation and enhance vascular function. In recent literature, these effects were related to the upregulation of autophagy. Autophagy is a cellular housekeeping mechanism and autophagy deficiency is detrimental in the development of CVD. Thus, inducing autophagy is likely to be an effective preventive strategy. Olive polyphenols were identified as natural autophagy inducers, but further research is needed to define the contribution of this mechanism to their antioxidant and atheroprotective effects. Therefore, we will determine the most potent autophagy-inducing olive polyphenol in endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) and define the underlying molecular mechanisms linked to autophagy. The most potent polyphenol will be selected for further in vivo analysis, in which we will identify its effects on the functionality of healthy blood vessels, CVD prevention and oxidative stress, with a special focus on the role of EC and VSMC autophagy. This project will give insight in the mechanism of action of olive polyphenols and is an important step towards the implementation of olive polyphenol-based nutraceuticals for the prevention of CVD." "From hit to lead: inducing basal autophagy for treating cardiovascular disease." "Pieter-Jan Guns" "Physiopharmacology (PHYSPHAR)" "Autophagy is a normal physiological process that maintains intracellular homeostasis by degrading unnecessary or dysfunctional cellular components in lysosomes. This way, autophagy supports cell survival in unfavourable conditions and represents a reparative and life-sustaining process. Impaired autophagy is increasingly recognized as a hallmark of aging and of multiple human pathological conditions, including cardiovascular disease. Inducing autophagy could be a game-changer in the treatment of cardiovascular disease, but the potential of autophagy inducing drugs has not been realized yet due to the absence of potent and selective tool compounds. The current proposal will start from a number of hits identified in a high-throughput screening and will further validate these lead candidates through a series of in vitro and in vivo studies focussing on vascular biology." "Research in the domain of toxicology" "Alexander van Nuijs" "Toxicological Centre" "The planned research activities are based on my experience accumulated in the field of bioanalysis. The research plan consists of further developing of my current research lines, as well as introducing new research directions in order to support a UA frontline (drug discovery and development) and emerging research domain (proteomics, genomics, and metabolomics). I hypothesize that biomarkers present in wastewater produced by communities provide objective information on spatio-temporal patterns on their lifestyle, disease and exposure to toxicants (= wastewater-based epidemiology). This will be tested through innovative research that will (i) select small-molecule biomarkers related to lifestyle, disease and exposure to toxicants; (ii) develop state-of-the-art bioanalytical assays to measure biomarkers in wastewater; and (iii) interpret the data with advanced statistical and modelling techniques. The aim is to implement wastewater-based epidemiology as a flexible and holistic tool to provide objective and timely information on the health status of whole communities. To allow long-term monitoring of alcohol and drug consumption in forensic contexts (e.g. driver's license regranting, organ transplantation procedure), I will continue my research to explore the use of keratinous matrices, i.e. hair and nails. I will investigate differences in hair and nails in accumulating substances. Furthermore, I want to establish reference values for biomarkers of drugs and alcohol in nails to be applied in a forensic context (as they also exist for hair). Therefore, studies with a high number of participants will be performed to correlate alcohol and drug biomarkers. I further plan to establish a robust bioanalytical platform to be used in the preclinical testing and pharmacokinetic profiling of candidate pharmaceuticals. I will focus on optimizing in vitro bioanalytical techniques for the evaluation of aspects of absorption (rapid equilibrium dialysis), distribution (protein binding assays) and metabolism (incubations with liver preparations) of xenobiotics. Furthermore, dedicated bioanalytical assays can/will be developed for the pharmacokinetic modeling of candidate pharmaceuticals (clearance, distribution volume,…). I aim to develop state-of-the-art bioanalytical procedures for metabolomics that combine multi-dimensional chromatographic and ion mobility separation techniques coupled to mass spectrometry (2D-LC-IM-QTOFMS) for the simultaneous detection of endogenous metabolites in a variety of biological matrices (cells, tissues, body fluids). This innovative technological platform will be used to (i) improve currently available bioanalytical metabolomics protocols in the Toxicological Centre for lipids and amino acids and (ii) develop innovative bioanalytical platforms to study subsets of the metabolome that are relevant for cardiovascular and inflammatory diseases: eicosanoids and steroids. Efforts will be made to optimize pre-analytical aspects including metabolism quenching and analyte extraction. These platforms can eventually be used in larger research contexts where metabolic information is crucial and can be integrated with other omics techniques to understand pathophysiological processes." "Research in the field of pathophysiology." "Tom Vanden Berghe" Pathophysiology "Unprotected iron can rust due to the attack of oxygen. Similarly, in our body, oxidative stress can kill cells in an iron-dependent way, which can give raise to organ injury or degeneration. This newly discovered type of cell injury or necrosis is referred to as ferroptosis. The study of how this type of cell death works at the molecular levels gains a lot of interest, due to its assumed high clinical relevance. On the one hand, our research focusses on using ferroptosis or 'biological rust' to eradicate cancer such as neuroblastoma using nanomedicinal approaches. On the other hand, blocking ferroptosis using small compounds is intensively investigated in an attempt to interfere with e.g. acute organ failure in intensive care patients or patients with chronic degenerative diseases. This work is imbedded in an interdisciplinary approach and occurs in collaboration with experts and physicians in the field." "Validation of the oxadiazolone isostere as a carboxylate replacement in caspase inhibitors: approaches involving Strecker-based synthetic methodology development and on-target strategies." "Pieter Van Der Veken" "Medicinal Chemistry (UAMC)" "Caspases are intracellular, aspartate selective cysteine proteases. Given their central role in cell death and inflammation, caspases have been studied intensively as drug targets to date. In spite of impressive preclinical results and significant investment in clinical evaluation, no caspase inhibitors have so far been approved as drugs by FDA or EMA. Two important reasons therefore are commonly cited: (1) The large structural homology of caspases that complicates the identification of selective compounds. (2) The limited biopharmaceutical quality of most compounds. Many contain an irreversible covalent warhead function that can potentially induce off-target effects. Most inhibitor families also contain a free carboxylate. Both the ionic character of this group and its potential for toxic metabolite formation, most probably discount critically on the permeability and ADME-Tox properties of inhibitors. Preliminary work at UAMC has identified the oxadiazolone moiety as a useful isosteric replacement for carboxylates in caspase inhibitors. Research in this proposal will validate this finding by introducing an oxadiazolone group in several relevant classes of caspase inhibitors. In addition, synthetic methodology based on the Strecker reaction will be elaborated. The latter will allow efficient access to caspase inhibitors with less reactive warhead types. Finally, drug discovery methodology will be developed that should allow ""on-target"" synthesis using caspase 1 as a model." "Characterization of occurrence, metabolism and contribution to human exposure of new chemicals present in the indoor environment." "Adrian Covaci" "Toxicological Centre" "Chemicals are typically used in various products because of their beneficial properties, e.g. flame retardants, plasticizers, etc. However, most of them leach into the indoor environment and may provoke health disorders (e.g. endocrine disruption). With this proposal, we aim to enrich the fundamental knowledge related to new chemicals of concern by studying their occurrence in the indoor environment. Furthermore, we will characterize their biotransformation in humans by in vitro experiments with human liver subcellular fractions. We will then assess for the first time the resulting human exposure to these new chemical entities by investigating human biological samples (urine and handwipes) for the presence of these chemicals and their biotransformation products. Resultingly, we will propose a range of new biomarkers of exposure to be used in future biomonitoring studies. The detection of the newly identified chemicals (and/or their metabolites) in human biological samples will confirm occurred exposure and will assist in the prioritization for further investigation of other aspects, such as toxicity and possible health effects." "Drug delivery systems and in vivo efficacy of the serine protease inhibitor UAMC-00050 in a preclinical model for irritable bowel syndrome." "Benedicte De Winter" "Laboratory of Pharmaceutical Technology and Biopharmacy, Medicinal Chemistry (UAMC), Laboratory Experimental Medicine and Pediatrics (LEMP)" "The serine protease inhibitor UAMC-00050, previously shown efficacious in 2 preclinical IBS models after systemic administration, will be formulated for oral and rectal administration. The drug delivery systems will be tested in the rat IBS model for dose determination, efficacy, time of administration and serum concentrations, a crucial step towards further valorization of the compound." "Carboxypeptidase U - a new drug target for the improvement of treatment in acute ischemic stroke." "Medical Biochemistry" "Thrombolysis with tissue plasminogen activator remains the only approved pharmacological treatment for acute ischemic stroke, AIS. Besides the narrow therapeutic time window, its use is limited by its efficacy: in up to 50% of the treated patients, timely recanalization is not achieved. Moreover, administration involves serious side effects such as intracranial hemorrhage and neurotoxicity. Consequently, the search for new agents for improvement of AIS treatment is urgently needed. Research has demonstrated that the enzyme carboxypeptidase U (CPU, TAFIa) is an important player in thrombus lysis. After activation from its precursor proCPU, the released CPU is able to potently attenuate fibrinolysis. Consequently, inhibition of CPU activity is a novel approach to enhance fibrinolysis. We want to explore the involvement of this enzyme in AIS in more detail. The usefulness of CPU as a diagnostic marker to discriminate ischemic from hemorrhagic stroke and the relationship of CPU with clinical outcome and thrombolytic treatment efficacy will be investigated. We plan to optimize the Thrombodynamics assay in order to assess the effect of CPU-inhibition on clot lysis during thrombolysis. Furthermore, in a preclinical setting, we will evaluate the effect of CPU inhibition in an experimental stroke model in rats. This research will provide essential information on the role of the CPU system and the usefulness of CPU inhibitors as potentially efficient and safer treatment of AIS."