Protein phosphorylation is a process that activates biological and cellular pathways. This process is carried out by enzymes called protein kinases that add phosphates to proteins. Protein kinases are crucial drug targets involved in the onset and progression of human diseases such as cancer and Alzheimer’s disease. Growing evidence suggests that protein kinases are regulated spatially and temporally by the phase separation and condensation of biomolecules into cellular organelles that are not surrounded by membranes. The molecular mechanisms that govern the action of kinases in phase-separated condensates, however, are unknown. Knowledge of these mechanisms is crucial for the development of better drugs that target protein kinases. To address this challenge, I have designed an interdisciplinary project that goes well beyond the state-of-the-art to explore in atomic detail the action of protein kinases intimately involved in Alzheimer’s disease and the abnormal phosphorylation of the protein Tau. The PhaseKin project aims to (i) reveal the specificity and reaction kinetics of protein kinases inside phase-separated condensates in vitro and in cells, (ii) decipher the dynamic conformational landscape of the protein kinases MARK2 and GSK3β by advanced NMR methods that will grant unprecedented detail on their modes of regulation, (iii) disentangle changes in population distributions and rates of interconversion between structurally distinct kinase states inside condensates, (iv) unravel the physicochemical basis of kinase drug partitioning into condensates. The highly innovative nature of the project is devised to delve into the heart of protein kinase function and to revolutionize our knowledge about the chemistry of drug-kinase interactions. Findings from the PhaseKin project will provide critical guidance in the development of more efficacious and specific drugs which target protein kinases.

There is growing evidence that currently used toxicological assessments of chemicals fail to fully capture their actual biological activity. While chemicals are routinely tested for acute toxic effects, often at high concentrations, potential function-modulating effects at low concentrations are often underexplored. The inclusion of up-to-date high-resolution methods in the toxicological screening praxis would allow instead to fully capture the complex bioactivity profile of these compounds, which depends on numerous aspects including bioavailability, route of exposure and individual susceptibility. While the last 100 years have seen an enormous number of chemicals introduced into our daily life, of which a large number are well-known to persist in the environment, a sufficient understanding of their bioactivity potential is lacking. Also based on growing evidence, it is not far-fetched to hypothesize that some of these industrial chemicals could be partly responsible for the constant increase of non-communicable diseases including autoimmune diseases, chronic inflammatory diseases but also cancer or (neuro)degenerative diseases. Perfluoroalkyl and Polyfluoroalkyl Substances (PFAS) are a class of xenobiotics with proven strong toxicity at high-level exposure, while little is known about their bioactivity at concentrations commonly detected in human samples. Considering their high stability and ubiquitous occurrence in the environment and human body, exposure to PFAS has reached a pandemic scale. I postulate that it is of outmost importance to finally dissect the potential of these compounds to molecularly interfere with organ functionalities at the single-cell level using state-of-the-art high-resolution omics technologies. Supported by an interdisciplinary training in pharmacology, systems immunology, omics, animal and cellular models, and bioinformatics, I represent a prime candidate to successfully develop this project and clarify the bioactivity of PFAS.

Proteins are among the most essential molecules of life. In order to fulfil their native function(s) they need to adopt a well-defined three-dimensional structure. However, under some circumstances, proteins can misfold and/or form toxic amyloid structures, which are the hallmark of a range of diseases including type 2 diabetes and neurodegenerative diseases. In particular, the small pre-synaptic protein, alpha-synuclein (AS), whose aggregation is the hallmark of Parkinson’s disease, can adopt in vivo and in vitro an intrinsically disordered conformation in solution and an alpha helical state when bound to membranes; the equilibrium between these two conformations has been shown to be important for its proposed native function, e.g. synaptic plasticity, and to modulate its kinetics of fibril formation. Here, I propose to investigate the physiological factors responsible for the switch between functional and deleterious interactions between membrane bilayers and AS due to ageing or disease using an innovative combination of biological, structural, thermodynamic and kinetic studies. In particular, I propose to use both synthetic lipid model systems and isolated synaptic vesicles to study the effect of ageing and the presence of lipids associated with Parkinson’s disease pathology on the nature of the interaction between AS and lipid bilayers. The synaptic vesicles will be isolated from the brain of mice at different ageing stages, as well as from mice carrying gene modifications or knock out related to PD. The interaction between AS and the vesicles will be studied using a range of biophysical techniques including circular dichroism, fluorescence and nuclear magnetic resonance spectroscopy, Atomic Force and Electron Microscopy. The aim of this study is to establish a thorough understanding of the interplay between changes in lipid composition and increased propensity of protein aggregation.

Social anxiety disorder (SAD) – the most common type of anxiety disorders – has severe emotional, financial and social consequences for patients, their families and their social network. Affecting about 7% of Europeans at some point in their life, SAD produces substantial societal cost due to long lasting treatments and extensive periods of reduced working capacity. To address this important problem, the project will establish an innovative system for treating SAD by exploiting the proven therapeutic potential of advanced Virtual Reality (VR) technology. SAD is predominantly treated with cognitive behavioural therapy, in which patients are repeatedly exposed to fear-provoking, social situations. However, patients often have to wait very long before treatment becomes available, exposure is rarely applied with the necessary intensity due to resource limitations, and objective measurement of treatment efficacy and dynamic adjustment of therapeutic scenarios is hardly possible. These problems can be overcome with VR based, controlled exposure to virtual renditions of fear provoking situations, which has been shown to surpass traditional therapies in terms of treatment efficacy, patient acceptance, and economic efficiency. Given that advanced VR technology has become very affordable, a widespread application of VR for the treatment of SAD is now within reach. The proposed project will fuse the applicant’s extensive VR expertise with established principles for VR based therapy to create an innovative system for the treatment of SAD. This system will, for the first time, combine interactive 3D environments with external physiological measures (e.g eye tracking data) and real-time data analyses techniques to provide a precise, flexible and time and cost-efficient therapeutic approach. Established collaborations with clinical partners will allow for continuous testing and refinement of the system, thus ensuring that market readiness can be achieved within the funding period.