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Fostering high scientific quality in protein research in Eastern Slovakia

Periodic Reporting for period 1 - CasProt (Fostering high scientific quality in protein research in Eastern Slovakia)

Reporting period: 2020-10-01 to 2021-12-31

Slovakia belongs to the lowest R&I performing countries in the EU; moreover, substantial economic differences exist between individual Slovak regions. About 70% of the Slovak GDP per capita is produced in Western Slovakia. Eastern Slovakia contributes only 14% to the Slovak GDP. Project CasProt with the coordinating organization, Pavol Jozef Šafárik University in Košice (UPJŠ), the second-best classical university in Slovakia, will provide excellent support for scientific development in one of the least prosperous regions of Slovakia (Eastern Slovakia). A general objective of CasProt project is to significantly strengthen the scientific excellence in protein science at UPJŠ by linking it with two internationally leading institutions, Technical University of Munich (Germany) and University of Zürich (Switzerland).

Success of this project will unavoidably lead to a higher reputation and enhanced networking channels of UPJŠ. We believe that UPJŠ will become a leader in the field of protein science in Slovakia after the project implementation, with the potential to attract and incorporate recognized as well as early-stage researchers into UPJŠ and, in the long-term, slow down or even reverse the brain-drain from Slovakia.
Successful outcomes of the project's scientific activities will enable a new strategy for searching for new potential drugs targeting pharmaceutically relevant GPCR and developing new biotechnologically and pharmaceutically interested biomacromolecules. The contribution of the susceptible molecular technology platforms (single-molecule techniques and display methods) established in Košice can significantly reduce the necessary costs for expensive drug-development stages such as high-throughput screening of molecules as potential drugs targeting specific signaling pathways of GPCRs. Such cost reduction for the R&D process should be projected in a decreased price of the co-developed drugs and hence may find a broader use among less privileged members of the society.
The connection of the project actions with the educational process at UPJŠ will constitute a platform for creating a unique ecosystem of interdisciplinary-oriented education in bioscience at UPJŠ and, in the future, in Slovakia.

Concrete main objectives are:
(i) Reinforcement of scientific capacity and raise the research profile of the coordinating institution UPJŠ in the field of protein science
(ii) Identification and integration of the experienced and young scientists/early-stage researchers at UPJŠ
(iii) Enhance and stimulate the technological capacity of the coordinating institution UPJŠ
(iv) Strengthening the research management and administration skills at UPJŠ
Collaboration between our groups at UPJŠ and groups at partner universities, the group of Prof. Plückthun at the University of Zürich and the group of Prof. Rief at the Technical University of Munich, has been successfully established. We used a time window between two pandemic waves of COVID-19 for researchers from UPJŠ to carry out their study stays at our partner universities: 2 PhD students (2x 2 months) and 1 senior researcher (1x 1 month) at the University of Zürich and 1 PhD students (1x 2 months) and 1 senior researcher (1x 0.5 month) at the Technical University of Munich. The increased collaboration between UPJŠ and partner universities led to two joint publications.
The collaboration with the top laboratories in the field has led to a relatively steep increase in the number and quality of papers published by the members of the CIB-TIP UPJŠ laboratory in the field of protein science. In fact, while during 2019 and 2020 there were 8 (6 Q1 and 2 Q2) and 12 (7 Q1 and 5 Q2) papers published, respectively, in 2021 20 papers (15 Q1, 4 Q2, and 1 Q3) were published.
The plan for the conference attendance in the first half of the project has been accomplished. We attended three recognized conferences (e.g. EBSA meeting in Vienna, July 2021) and altogether 11 senior and early-stage researchers participated on these conferences.
Support of our research by project CasProt and projects from the EU structural funds OPENMED and BioPickmol as well as by projects supported by Slovak grant agencies APVV and VEGA had a synergic effect on obtained results that have been presented in five lectures and nine posters on foreign scientific conferences.
New technologies and methods have been established in the laboratories of UPJŠ, such as the protein evolution method – ribosome display. We also created the kappa opioid receptor (hKOR) library and performed the first analysis of the GPCR library by ribosome display. We selected several hKOR candidates that will be subsequently analyzed experimentally. Practical skills of utilizing a new method of the expression and purification of eukaryotic proteins based on the baculovirus expression system (BEV) have been obtained during the stay at UZH. Adapting BEV enables purifying GPCRs and other different eukaryotic proteins, membrane proteins or protein complexes in UPJŠ laboratories.
Short-stays at TUM taught us a basic knowledge and technical aspects of the preparation of so-called nanodiscs that, combined with the expression of functional GPCRs, provide a unique opportunity to study biophysical properties, such as dynamics and stability of integral membrane proteins in their natural environment. In the case of GPCR protein, the implementation of this system and the optimization of steps will lead to the development of comprehensive procedures for monitoring GPCR dynamics using single-molecule techniques.
Thanks to CasProt support, we have established a new collaboration with the top laboratory in the field of rational design - the group of Prof. Damborsky at Loschmidt laboratories at Masaryk University in Brno in the Czech republic. Consequently, based on this newly established collaboration, we have submitted the bilateral scientific project within the APVV call on the topic related to the utilization of ribosome display in improving properties of thrombolytics based on staphylokinases with vast potential in clinical application.
In summary, the CasProt project can significantly contribute to the long-term sustainability of excellent research, technology transfer, and raise research profile of UPJŠ, specifically in the ambitious fields of single-molecule protein folding and stability, directed evolution of proteins and potentially, drug delivery system development. Based on this fact, the CasProt project will contribute to the enhancement of the R&I performance of Slovakia and significantly subsidize the creation of highly competitive research institutions in the less developed region (Eastern Slovakia) and by this means strengthen the economic potential and social cohesion of this region. All this can reduce the regional disparities existing in Slovakia, which is one of the main priorities of the Slovak government as well as one of the primary purposes of the WIDESPREAD-05-2020: Twinning call.
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