HMGCR (3-hydroxy-3-methylglutaryl-coenzyme A reductase) is an enzyme that plays a crucial role in cholesterol biosynthesis. Research on HMGCR is important for numerous patients with atherosclerosis because inhibiting the enzyme with statins has been shown to reduce the risk of cardiovascular events such as heart attacks and strokes. Despite being generally well tolerated, statin intolerance is reported in 5-20% of patients, leading to reduced regimen adherence and statin therapy duration. In addition, HMGCR may also have a role in the development of diabetes. The goal of the Project is to discover molecular pathways linking excessive inhibition of HMGCR and hepatotoxicity, myopathy and neurological deficits. We will map the molecular effects of HMGCR deficiency in a unique mouse Hmgcr-KO model, thus advancing the understanding of mechanisms behind the statin treatment-induced tissue-specific adverse effects. We will specifically explore the role of HMGCR inliver cells and its involvement in the mechanisms of mitochondrial fatty acid metabolism. All this will help to evaluate the hypothesis of HMGCR as a crucial enzyme for maintaining cellular energy metabolism balance and to drive discovery of innovative approaches to prevent and treat cardiometabolic diseases.

Alzheimer’s disease (AD) is an age-related neurodegenerative disorder responsible for about 2 million deaths per year. Despite tremendous progress in basic research within the last years, its efficient treatment and diagnostic tools are still lacking. The previous studies have gathered sufficient evidence of a causative role of the aggregates of two proteins - amyloid beta and tau - in the AD pathogenesis. Both of these proteins can form highly toxic oligomeric species, which are the primary suspects of AD-related neurotoxicity. However, due to experimental difficulties the detailed structural and functional characterization of the oligomeric forms has been a big challenge. In the proposed project we aim to build on cutting-edge technologies and novel approaches to obtain atomic-level structures and investigate interactions of the amyloid beta and tau oligomers. Thus, this multidisciplinary project will apply (I) cell-free protein expression and purification protocols for incorporation of various selectively 13C, 15N and 19F labeled amino-acids; (II) microfluidics in order to generate size-controlled oligomers (III) solid-state NMR (ssNMR) at fast magic-angle spinning (MAS) regime tailored for 1H and 19F detection schemes. The anticipated outcome of this will be a unique combination of approaches to study amyloid aggregates, which can be further used and adapted for studying other protein assemblies. The project results will form the basis of innovation in the treatment of AD as well as other tauopathies.

Heart failure with preserved ejection fraction (HFpEF) is a syndrome associated with high morbidity, mortality, and increasing prevalence. Development of HFpEF is driven by metabolic inflammation and alterations in energy metabolism with concomitant accumulation of lipotoxic metabolism intermediates. Long-chain acylcarnitines (LCAC) are fatty acid metabolism intermediates which at elevated levels impair mitochondrial bioenergetics and activate pro-inflammatory signaling pathways. This project aims to assess the role of LCAC in HFpEF development and evaluate LCAC pool-lowering therapy as a potential treatment for HFpEF. HFpEF will be induced in mice by treating them with high fat diet and L-NAME in their drinking water. LCAC pool will be decreased by treatment with meldonium. To characterize HFpEF development, heart function, energy metabolism and physical performance in vivo, function of isolated skeletal muscle, mitochondrial bioenergetics will be assessed. Additionally, the effects of concomitant administration of an SGLT2 inhibitor with meldonium will be studied in the same experimental setup. The role of LCAC on the development of HFpEF will be studied in trimethyl-lysine hydroxylase knockout mice which do not express enzyme essential for the LCAC synthesis and have a very low LCAC levels. Obtained results will provide new knowledge about the role of LCAC in the development of HFpEF as well as lay grounds for discovering novel drugs for treatment of HFpEF.

The candidiasis disease is one of the more severe fungal infections with particularly high mortality amongst immunocompromised patients. It is caused by the opportunistic fungus Candida albicans (C.albicans) that is quite resistant to common antifungal medicines. The cell wall of C.albicans has a key role for protection and interaction of fungus, making it a promising target for new antifungals. However, the structural organization at atomic level for the intact C. albicans cell walls remains understudied. Here we are proposing a project, which will focus on the cell wall molecular organization studies at atomic level for the intact C. albicans fungus using non-destructive solid-state nuclear magnetic resonance (ssNMR) spectroscopy. We will elucidate the intact C.albicans cell walls by exploiting modern 1H and 19F detected ssNMR technologies at fast magic angle spinning regime. During this interdisciplinary project we plan to improve the selective 1H, 13C, 15N labeling schemes and create a novel biochemical 19F incorporation in the C.albicans cell walls. In combination with development of new 1H and 19F detected ssNMR techniques, we will elucidate the structures of main polysaccharides, lipids and proteins, allowing to decipher the supramolecular arrangements. The anticipated findings in this research project will provide a precise structural model of the intact C.albicans cell walls, paving a way for discoveries of new antifungals.