Structure determination of amyloid oligomers, the pathogenic species in Alzheimer ́s disease using fast MAS NMR and microfluidics (2021-2023)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.
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Moving NMR infrastructures to remote access capabilities (2022-2025)
Remote NMR (R-NMR) is 3 years project by 26 European NMR groups/entities that aims to design and deliver a platform for remote access capabilities to Nuclear magnetic resonance (NMR) Research infrastructures (RIs). Nuclear magnetic resonance spectroscopy (NMR) is one of the major analytical methods applied in all chemical, physical, biological, and medical sciences. NMR’s leading role stems from its analytical power in terms of molecular resolution, quantification, reproducibility, and broad application envelope. It requires sophisticated and expensive equipment, operated by scientists with diverse background ranging from service-oriented researchers to highly trained experts. The aggregated capital investment in European NMR facilities, operating at local, national, and European level, exceeds 500 Million Euro. The NMR community maintains excellent networking between sites and serves a broad community within the focal points of European research interests. With the application Remote-NMR (R-NMR), the Consortium wish to establish remote access for all NMR users throughout Europe. While ordinary NMR applications are routine in every university and in industry, more specialized applications are performed in dedicated RIs offering access and help to local and outside users. During the pandemic, NMR infrastructures slowed down their operations to variable degrees, and particularly access by remote users dropped significantly, raising the need to establish standardized procedures for remote access to improve their resilience to adverse external factors. This project aims to establish an inclusive network of NMR-infrastructures throughout Europe, to survey if and how remote access can be made possible according to the needs of the community, and to implement GDPR at facilities and sample shipment procedures. Routines for remote NMR-usage will be also established, including dissemination of research and teaching protocols, archiving of data, and sample shipment. The overall CO2 footprint of the operation of the consortium will be evaluated, as well as its reduction due to the reduction of travels.
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This project has received funding from the European Union’s Horizon Europe research and innovation programme under grant agreement no. 101058595.
Integrative structural biology of pathological tau protein, an appealing therapeutic target for Alzheimer’s disease modifying drugs (2020-2025)
There is an enormous unmet medical need to find efficient methods of prevention, diagnosis and disease-modifying therapies for tauopathies, including Alzheimer’s disease. The common molecular denominator of tauopathies are pathological forms of tau protein. Tau pathology relates to conformational changes during oligomerization and assembly resulting in toxicity. Given its role in the pathogenesis, conformationally altered and assembled tau would be a promising molecular target for disease-modifying therapy. However, the field is still lacking deeper understanding of tau structural changes in the course of assembly and their inducers on the pathway towards the pathological forms of Tau; therefore, pharmaceutical development is hampered. The main aim of the InterTau project is the detailed structural and biophysical characterization of tau protein and its variants in monomeric, oligomeric and fibril states relevant for AD and other tauopathies. The InterTAU consortium is composed of a clinical-stage biotech company pursuing development of anti-tau immunotherapy and academic partners with cutting-edge methodologies suitable for functional and structural characterization of tau assembly by solution and solid-state nuclear magnetic resonance (NMR), cryo-electron microscopy and cellular assays corroborated by bioinformatics. The mutual transfer of complementary expertise envisaged in the project will facilitate academic outcome and biotechnological development. Specific expertise will be transferred from three institutions in North America and one institution from Argentina. The results of InterTAU will be directly translated into innovation in pharmacological development through the non-academic partner. The platform for sharing knowledge will be a foundation of sustainable cooperation beyond the InterTau project.
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Novel 19F NMR based methods for structural biology of pathological tau protein, an appealing Alzheimer ́s disease therapeutic target (2020-2022)Alzheimer’s disease and other tauopathies pose a formidable clinical challenge as well as several unmet medical needs to find efficient tools and therapies for their prevention, diagnosis and treatment. All tauopathies are associated with aggregation of pathological forms of the tau protein leading to neurofibrillary tangles in the brain, which makes it a promising target for therapy. However, there is still lack of understanding of the factors and conformational changes during tau oligomerization and assembly that result in specific pathologies. The proposed project aims to unravel the structural and dynamic details of the tau assembly pathway towards the pathological forms by establishing innovative approaches based on solution and solid-state nuclear magnetic resonance (NMR). We will build on cutting-edge methodologies and novel approaches for synthesis and incorporation of 19F labelled amino acids and high-sensitivity NMR experiments to obtain structural and functional characterization of the tau protein and its variants in monomeric, oligomeric and filamentous states relevant for disease. The project will heavily benefit from collaboration with partners of the H2020-MSCA-RISE InterTAU project. The project results will form the basis of innovation in the treatment of Alzheimer’s disease and other tauopathies. |
Chemically modified artificial spider silk (2019-2022)
Spider silk is biocompatible, biodegradable, strong and elastic, which makes it ideal for biomedical applications. However, harvesting of silk from spiders is not feasible for large scale industrial purposes and current biotechnological approaches face problems with low recombinant protein yields as well as difficulties reproducing the native mechanisms of fibre spinning. The aim of the present project is to develop a novel method based on bioconjugation with polyethylene glycol for obtaining chemically modified artificial spider silk in a biomimetic way. The developed method would have several advantages in comparison to a purely biotechnological approach as it would ease recombinant spider silk protein (spidroin) expression in bacteria by circumventing the requirement for all three spidroin domains, allow crosslinking between spidroins as well as easy tuning of material properties by varying the bioconjugation reaction conditions. Successful implementation of the project will contribute to the development of several defined specialization areas of the Latvian Smart Specialization Strategy (RIS3) – biomedicine, medical technologies, biopharmacy and biotechnology. The project results will be applicable to development of new biomaterials with potentially broad applications in biomedicine and nanotechnology and thereby contribute to transformation of economy of Latvia towards high added value products. |
From molecules to crystals: molecular self-assembly in crystal nucleation from solution (2018-2020)
Crystallisation is the most common method of chemical compound isolation from solution, and it is extensively used in chemical, food and pharmaceutical industry. Nevertheless, early stages of crystallisation are still poorly understood. Nucleation – the formation of the first stable embryonic structure of crystal in supersaturated solutions, plays a central role in determining the structure and size distribution of the final crystals, therefore, understanding the fundamental nucleation aspects are crucial to the control of crystallisation processes. |
Understanding prion peptide fibril-induced aggregation of prion protein (2017-2019)
Prion-like spreading may be employed in a number of fatal neurodegenerative disorders, including such as Alzheimer’s and Parkinson’s diseases. Understanding all possible mechanisms of such spreading would be a big step towards curing these diseases. |
