Research projects

Haemocoagulation and fibrinolysis

The aim of the project is to describe the relationships between the individual components of the coagulation cascade and their influence on pathophysiological conditions (e.g. oxidative stress). We study coagulation from the activation of serine proteases, through the dynamics of the formation and structure of the resulting fibrin network, to fibrinolysis. We use not only conventional coagulation methods but also new approaches such as mass spectrometry, electron microscopy and structural biology methods. The findings are applied in the development of new diagnostic approaches for coagulopathies (e.g. deep vein thrombosis, pulmonary embolism, myocardial infarction, thrombotic complications in COVID-19).

Characterization of fibrinogen disorders

In patients with suspected dysfibrinogenemia and hypofibrinogenemia from all over the country, we investigate all exons of fibrinogen-encoding genes (FGA, FGB and FGG). If a mutation is found, we investigate its effect on the alteration of fibrinogen structure and function using specialised tests (polymerisation curves, quantification of fibrinopeptide cleavage by HPLC, SEM, LC-MS/MS, structural modelling). Overall, more than 100 cases from more than 70 families have been identified so far at our institution, of which 16 mutations are newly discovered.

Computer modelling

Theoretical structural biology methods such as molecular modelling and molecular dynamics are mainly used to study fibrinogen and its interactions with other molecules in the blood. Specifically, these include studying the effect of post-translational modifications and mutations on the structure and function of fibrinogen, describing the interactions of fibrinogen with thrombin and hemostatic snake venoms, and predicting the structure of portions of fibrinogen that could not be determined by experimental methods. We also use methods of sequence and phylogenetic analysis.

Metabolomics of plasma/serum/cell extracts in oncohematological and other diseases

Using a combination of liquid chromatography and tandem mass spectrometry (LC-MS/MS), we are engaged in the targeted analysis of low molecular weight compounds. Specifically, we study changes in the concentrations of metabolites of glycolysis, citrate cycle, energy pool, carnitine metabolism, tryptophan and selected amino acids. We found differences in patients with acute myeloid leukemia (AML) depending on the course of the disease. We also monitor markers of oxidative stress (malondialdehyde, uric acid, allantoin, etc.). For the analysis of protein carbonylation we use spectrophotometric determination.

Protein-protein interactions by surface plasmon resonance (SPR)

The aim of this work is to design a diagnostic SPR method and protein chip to more accurately stratify subgroups of oncohematological or other diseases. We have designed a protein (micro) chip that analyzes interactions between proteins associated with the pathogenesis of a given disease and the plasma proteome of patients. Another proposed Hsp70 trap (micro) chip uses five distinct subcellular chaperones of the Hsp70 family and is used to detect the damaged subproteome of patients. The protein (micro) chip has already demonstrated the ability to discriminate between different subgroups of myelodysplastic syndrome (MDS) and healthy donors, while the Hsp70 trap (micro) chip has demonstrated the ability to discriminate between low- and high-risk MDS patients. Identification of interacting proteins by mass spectrometry revealed other suitable candidates to increase the diagnostic sensitivity of the protein chip.

Proteomics of blood cells and other blood components

The aim of this work is to elucidate physiological and pathophysiological processes in selected diseases or to use the findings in diagnosis or treatment monitoring. The main focus is oncohematological diseases (e.g. myelodysplastic syndrome), but also other diseases (e.g. cardiovascular diseases). Another area of research is the monitoring of the interaction of artificial (polymer) surfaces with blood, using knowledge in a wide range of areas from the design of diagnostic tools to biomedicine (tissue engineering, implants, etc.).