National Science Centre

The aim of the project is to investigate the biological effects of trimethylamine oxide (TMAO), which is produced by intestinal bacteria and may affect the normal function of the heart and the entire circulatory system. Heart failure is one of the most important cardiological causes of death, numerous hospitalizations, disability and reduced quality of life. Recent studies indicate that TMAO, a metabolite of intestinal bacteria, may play an important role in cardiovascular disease. In recent years, there have been reports that an increase in the concentration of trimethylamine oxide (TMAO) in the blood is associated with a higher risk of myocardial infarction, stroke and death. The increase in TMAO levels may be due to diet, impaired gut function, or changes in the composition of the gut flora. It is known that the concentration of TMAO in the blood increases after eating foods containing L-carnitine and phosphatidocholine, which are found in large amounts, for example, in red meat. It has been proposed that TMAO may constitute a long-sought link between diet and cardiovascular disease. On the other hand, a large number of previous studies on the role of TMAO in the biology of marine animals and physicochemical studies on the effects of TMAO on proteins indicate the protective effect of TMAO. Therefore, it can be assumed that the increase in the concentration of TMAO in the blood is not the cause, but the body's defense action, which is a protective mechanism against factors disturbing the proper functioning of the circulatory system. Another argument pointing to the beneficial effects of TMAO may be the fact that TMAO is found in very high concentrations in fish, especially marine fish, which are an important part of many diets considered beneficial to the circulatory system, such as the Mediterranean diet.

Neoplastic diseases constitute an important, still unsolved problem of clinical medicine and pharmacology of the 21st century. Therefore, intensive research is carried out into the development of new anti-cancer drugs that specifically destroy cancer cells, without causing negative side effects and with improved pharmacokinetic parameters. Interesting cytotoxic properties of several compounds from the group of benzo [b] furans and dicarboximides designed and produced at the Department of Medical Chemistry of the Medical University of Warsaw have been discovered in the Screening Laboratory of Anticancer Compounds operating at the Department of Bioorganic Chemistry of the Center for Molecular and Macromolecular Research of the Polish Academy of Sciences in Łódź. These compounds turned out to be cytotoxic to CML (K562), ALL (MOLT-4) and AML (HL-60) leukemia cells, while they are non-toxic to normal endothelial cells (HUVEC) and tumor adherent cells (HeLa and CFPAC). These compounds and their antitumor activity have been protected by Polish and international patents (EP13150611.5-1452, EP13176421.9, P.398193, P.400000). Aim of the research: Conducting advanced preclinical studies for selected benzo [b] furan and dicarboximide derivatives as potential anti-leukemic compounds, including testing their activity against hematopoietic stem cells from AML and CML patients and in vivo in animal models and understanding the molecular mechanism of action of these compounds in cells.

Aim of the research / research hypothesis: Thyroid neoplasms are the most common group of endocrine neoplasms, and their number is constantly increasing. Papillary thyroid cancer (PTC) is the most common, accounting for 85% of the total. The basis of its treatment is surgical resection of the thyroid gland with regional lymph nodes and complementary therapy consisting of the administration of radioactive iodine 1131. Some patients, however, are resistant to therapy with radioactive iodine. Due to the limited effectiveness of other forms of treatment, e.g. teleradiotherapy, chemotherapy, these patients die because of the aggressive course of the disease. Hence, it is necessary to search for new therapies. The protein product of the SLC5A8 gene was originally identified as an iodide transporter present in the apical membrane of thyrocytes and therefore called AIT (Apical Iodide Transporter). It is also a tumor suppressor and its expression is reduced in many cancers, including PTC. Most studies on its suppressor function concern the large intestine and are based on the ability of AIT to transport short-chain fatty acids. Only a few studies mention the modulation by SLC5A8 of other genes involved in the tumorigenesis process, and these are genes as fundamental as TP53, FASL, FASR, Bcl-2, TRAIL, TRAILR1 and 2, BIRC5 It is therefore surprising that no one has made the changes induced by SLCSA8 overexpression at the level of the whole transcriptome. There are only a few studies on the regulation of SLC5A8 expression, which directed our attention to the role of microRNAs in this process. MicroRNAs (miRNAs) are about 22 nucleotide non-coding RNA molecules that inhibit gene expression by binding to a complementary sequence in their transcripts. In our preliminary research we have shown that miR-181a-5β, -182-5p and -494 regulate SLC5A8 expression, and their increased level in PTC can lead to a decrease in SLCSA8 expression. We also showed the influence of microRNAs on the radioactive iodine transport process. Changes in the expression of microRNAs can be modulated by specific inhibitors, therefore we confirmed that transfection with a synthetic miR-181a-5p inhibitor increases the expression of SLC5A8 mRNA and we hypothesized that by inhibiting the activity of the identified microRNAs we would increase the expression of SLC5A8, which will lead to increased accumulation of radioactive iodine and to a change in the level of genes regulated by SLC5A8, which may increase the effectiveness of anti-cancer therapies.The aim of the project is to use specific microRNA inhibitors to restore the proper level of SLC5A8 in cells derived from thyroid cancer. We will conduct a double analysis of the effect of increased SLC5A8 expression on cell physiology: using next-generation sequencing, we will determine the total transcripts of cell lines in which the action of selected microRNAs will be inhibited (analysis of potential cytotoxicity of inhibitors). Next, we will examine the total transcripts of cell lines in which we overexpress the SLCSA8 gene (analysis of the role of SLC5A8 in the regulation of gene expression). Thanks to this analysis, we will identify the pathways in which SLC5A8 exerts its suppressor role. MiR-181a-5p, -182-5p and -494-3p are also overexpressed in other neoplasms, so we can call them onkomiRami. Therefore, it is even more justified to study the influence of their inhibition on changes taking place at the level of the entire transcriptome. The next step will be to compare gene expression changes caused by SLC5A8 overexpression and microRNA inhibition. Not only will this provide a list of target genes, but it will also bring us closer to the question of whether tissue-specific inhibition of these microRNAs could find therapeutic application in the future. Research method / methodology used: During the project implementation, we will create a plasmid expressing the coding sequence of the SLC5A8 gene and the so-called sponge for miR-181a-5p, -182-5p and -494, expressing a transcript with sequence complementary to 3 microRNAs, acting as a specific inhibitor of these molecules. Optimal transfection conditions will be determined by measuring the expression of SLC5A8 and at least one of its target genes described by using real-time PCR. After determining the optimal protocol, we will isolate RNA from the cells, prepare a cDNA library and subject it to next-generation sequencing (NGS). We will create lists of genes with altered expression due to SLC5A8 overexpression and (separately) microRNA silencing. We will then compare these lists with each other to determine changes caused by gene overexpression, changes caused by silencing of selected microRNAs, and consequently we will determine the potential utility of microRNA inhibitors in restoring SLC5A8 expression in PTC cancer. The impact of the expected results on the development of science, civilization, society: This will be the first project to characterize the influence of SLC5A8 on the expression of other genes at the level of the entire transcriptome. We will also determine the possibility of restoring its expression by microRNA inhibition. Importantly, thanks to the use of the NGS technique, we can identify other target genes useful from a therapeutic point of view for these microRNAs. We will compare the changes caused by overexpression of the SLC5A8 gene with those caused by silencing of selected microRNAs, which also regulate other target genes, including those involved in tumor suppression. Thanks to this, we will determine the potential utility of silencing selected microRNAs in patients suffering from papillary thyroid cancer.

The intestinal epithelial cells constitute the single layer of epithelium that forms a barrier that separates the intestinal lumen from the immunocompetent cells of the GALT system. The intestinal epithelium is now considered an important part of the gut immune system necessary for the induction and regulation of both non-specific and specific immune responses in the intestinal mucosa. Impaired immune function of epithelial cells may contribute to the development of inflammatory bowel disease (IBD) and food allergies. The main goal of the project is to evaluate the effects of bacteriophages (bacterial viruses) on the immune functions of the intestinal epithelium in vitro. Testing method / methodology used All experiments will be performed on the Caco-2 cell line. In the first stage of the study, the influence of bacteriophages on the expression of selected genes important for the immune functions of the intestinal epithelium will be assessed. Gene expression will be assessed using the RT2 Profiler PCR Assay kit. Significant bacteriophage-induced changes in gene expression will be verified by measuring the production of the corresponding proteins by ELISA (for soluble proteins) or by flow cytometry (for membrane proteins). The influence of the expected results on the development of science, civilization and society. According to our knowledge, this is the first study to assess the influence of bacteriophages on the immune functions of the intestinal epithelium. The implementation of the project will broaden the knowledge of the role of the intestinal microflora in the regulation of immune homeostasis in the intestinal mucosa and the pathogenesis of IBD and food allergies.

Asthma and chronic obstructive pulmonary disease (COPD) are the most common chronic respiratory diseases in our society. Their essence is inflammation in the lower respiratory tract, which leads to obstruction (narrowing) of the bronchi. The main symptoms of asthma and COPD are shortness of breath, wheezing heard when breathing, cough (dry or tough), and decreased exercise tolerance. The manifestation of the disease depends both on genetic predisposition as well as on environmental factors, such as allergens that favor the development of asthma and tobacco smoke, which is the most important cause of COPD. Both asthma and COPD are very complex and variable diseases. The diagnosis of asthma and COPD can be difficult because these diseases sometimes have quite similar symptoms. In practice, asthma and COPD are often two sets of respiratory diseases whose pathophysiology, clinical symptoms and treatment overlap to a large extent. One of the most important phenomena in the development and course of these diseases is the influx of various inflammatory cells into the respiratory tract. The presence and activity of specific groups of cells in the respiratory tract is closely related to the development of the symptoms of the disease. Typical cells for asthma are eosinophils, and in COPD the dominant role is played by neutrophils (neutrophils).However, this classic picture of airway inflammation ascribed to asthma or COPD is often difficult to observe as there are many mixed and overlapping phenotypes. The constant presence of an increased number of eosinophils and neutrophils in the lungs leads to the breakdown of these cells and the release of harmful messengers causing damage to the airways, leading to symptoms of the disease. As a result, the production of a large amount of secretions and irritation of the nerve endings are observed. For the patient, this means coughing, shortness of breath, worse respiratory test results and the need for frequent visits to the doctor due to respiratory infections. The cells in the airways are able to communicate thanks to the huge number of transmitters they produce. The cytokines produced by the epithelium: TSLP, IL-33 and IL-25 are important proteins that shape the allergic response of the organism. Thanks to these cytokines, it is possible to "talk" between the groups of cells that directly create inflammation in the airways. The role of TSLP, IL-33 and IL-25 is widely studied in asthma as a disease that is often allergic. The role and function of these cytokines in COPD are unknown. Atopy, i.e. a genetically determined complex of disorders of the immune system resulting from the body's reaction to a given allergen, is associated with TSLP, IL-33 and IL-25. In the light of recent scientific publications that emphasize the individualization of the diagnosis and treatment of obstructive respiratory diseases, it is important to understand the underlying biological reactions of allergic inflammation, which may occur in both asthma and COPD, especially in COPD with an allergic phenotype (eosinophilic inflammation in COPD). The presented project aims to assess the interactions and relationships between the types of cells forming the immune response of the respiratory system: macrophages, respiratory epithelium and dendritic cells through TSLP, IL-33 and IL-25 expression in atopic and non-atopic obstructive respiratory diseases. The project is to be carried out in a complex (three-layer) and advanced in vitro model that reflects the site of an active immune response in the human airways. The material for the study will be the respiratory epithelium isolated from nasal brush swabs, as well as macrophages and dendritic cells isolated from peripheral blood cells from patients with COPD, asthma and healthy people. Careful analysis of the biochemical reactions underlying allergic inflammation in patients with asthma and COPD will increase knowledge in the field of immunology and provide a better understanding of the mechanisms that govern the course of obstructive respiratory diseases.

The study design includes eight research hypotheses and 8 corresponding study objectives. The most important goals are outlined below. • Analysis of changes in lung ventilation indices and changes in gas exchange indices depending on the volume of fluid removed and changes in intrapleural pressure. • Examination of changes in the compliance of the lungs and chest walls and the related changes in the scope of respiratory work occurring during therapeutic pleural puncture and fluid evacuation. • Analysis of the relationship between changes in intrapleural pressure, dimensions and heart function assessed by echocardiography, and changes in blood concentration of type A natriuretic peptide and type B natriuretic peptide. pressure and ventilation measurements taken during the procedure. The examination will be performed in 60 patients with pleural fluid requiring therapeutic pleural puncture. Research before toracentesis. bodyplethysmography, spirometry, carbon monoxide diffusion capacity (DLCO), arterial blood gas testing, six-minute gait test, echocardiography, blood natriuretic peptide concentration.Assessment during thoracentesis: constant measurement of tidal volume and respiration rate, measurement of pleural effusion volume, measurement of pleural pressure, percutaneous monitoring of O2 and CO2 partial pressures. A catheter will remain in the pleural cavity to allow the fluid to drain and periodically measure the intrapleural pressure after the procedure is completed. Monitoring during 48 hours after surgery. In two groups of patients (undergoing physiotherapy aimed at accelerating the aeration of the lung and those who will not be subjected to physiotherapy), the tests before pleural puncture will be repeated at appropriate intervals. Pleural pressure measurements will be taken up to 48 hours after surgery. Thereafter, the catheter will be removed. Analysis. The signal analysis and visual multidimensional analysis of the relationship between parameters carried out with the use of proprietary computer programs will be an introduction to the formulation of physiological, medical and statistical hypotheses, and will also suggest possible modifications to the analysis and previously parameterized signals. The results of the study will allow to characterize the relationship between lung function indices, gas exchange indices and cardiac function, and changes in intrapulmonary pressure. Some of the studied relationships have never been studied before. The results of the study may change the standard of care during therapeutic pleural puncture and in the early period after its performance.

Parasites of the genus Trypanosoma have a unique cell physiology as they carry out important glycolytic and peroxisomal processes in one organelle, the glycosome. Since glycosomes do not contain genetic information, all enzymes active in this compartment must be delivered there post-translationally. A group of enzymes called peroxins (small peroxisomal proteins, PEX #) govern this transport. Of these, PEX5 and PEX14 play an important role in this biochemical pathway because complex formation between them is necessary for the import of proteins from the cellular matrix into the glycosome. For this reason, it is postulated that preventing the formation of this complex, e.g. by means of a small-molecule drug-like substance, could be an interesting strategy to fight diseases caused by Trypanosoma, as well as a method of studying biochemical processes taking place in glycosomes. Interprotein interactions are among the most difficult molecular targets. As with many other protein interactions, the interaction of PEX14 with PEX5 is predominantly hydrophobic and aromatic in nature, with only two shallow, exposed binding pockets located over a large contact area. As a result, to be able to compete for binding sites on the PEX14 surface, a "conventional" "drug-like" chemical molecule would need to be strictly lipophilic in nature. This, in turn, implies difficulties in keeping the pharmacochemical parameters within the desired ranges. Therefore, there is a need to look for alternative strategies to inhibit this difficult molecular target, other than "classic" small molecule chemicals. The aim of this project is to try to use alpha helix mimetics, oxopiperazine derivatives, as new inhibitors of PEX14 and PEX5 protein interactions with high antiparasitic efficacy and appropriate pharmacochemical properties. A multidisciplinary approach to achieving this goal will be based on research methods such as chemical synthesis, computer-based ligand design based on protein structure, as well as biophysical and cellular assays. Lessons learned from these studies may be important not only for the design of future potential drugs against tropical diseases, but also for a better understanding of the biochemical processes involved in glycosomes.

The project involves the production of a series of new chemical compounds, hydroxycoumarin derivatives, with high affinity for serotonergic receptors and the serotonin transporter. Compounds were designed based on a literature review and using molecular modeling methods. The structures of the compounds were planned on the basis of the previous research in which a pilot series of aryl-piperazinyl-propoxyl and butoxy derivatives of 8-acetyl-7-hydroxy-4-methylcoumarin was obtained, their activity profile and affinity for some serotoninergic receptors were determined. Initial results are optimistic and encourage further research. It was found that the introduction of an acetyl group at the 8-position of the coumarin ring contributed to a significant increase in affinity for the 5-HT1A receptor (Ki = 89.2 nM for the compound without an acetyl group to Ki = 0.8 nM for 8-acetyl-7- {4 - [4- (3-methoxyphenyl) -1-piperazinyl] butoxy} -4-methyl coumarin). Moreover, due to molecular docking, it was found that compounds of this type are stabilized in the receptor pocket mainly by the hydrogen bond between Ser190 and the acetyl group of coumarin. This binding has previously been reported to be important in the binding of (non-coumarin) ligands to the 5-HT1A receptor. Also important is the substitution of the 4-position on the coumarin ring with a methyl group and, of course, the selection of the appropriate linker and arylpiperazine derivatives having substituents in the ortho or ortho / meta position of the phenyl ring. Based on the above data, I decided to synthesize new arylpiperazinyl derivatives of hydroxycoumarins with the use of a three / four / five and six carbon linker and a number of amines. Organic synthesis is based on certain and proven chemical reactions. The compounds planned under the project will be obtained as a result of multistage organic syntheses, mostly with the use of a microwave reactor. The derivatives will be purified by crystallization or column chromatography methods, and their structures will be confirmed by1H NMR, 13C NMR, MS spectra, and by crystallographic methods for selected molecules. The main result of the presented project will be the purification and confirmation of the structures of planned chemical compounds with the expected pharmacological activity. The results will be published in international journals. We envisage the preparation of aryl-piperazinyl derivatives of 8-acetyl-7-hydroxy-4-methylcoumarin and 6-acetyl-5-hydroxy-4,7-dimethylcoumarin containing an acetyl group in ortho position to the coumarin hydroxyl group. The aryl piperazinyl derivatives of 5-hydroxycoumarin have never been described in the scientific literature or tested in terms of affinity to serotonin receptors. The project will allow to enrich the library of coumarin derivatives with new derivatives, and the obtained results will constitute the material of the project manager's habilitation dissertation. As the project is part of the medical chemistry framework, pharmacological tests will be necessary (apart from the presented application). The affinity for serotonergic receptors and the serotonin transporter will be determined based on in vitro studies (as part of scientific cooperation with the Department of Pharmacobiology, Faculty of Pharmacy, Jagiellonian University).

The subject of my scientific interests are molecularly imprinted polymers (MIPs). The process of creating MIPs known as molecular imprinting consists in the polymerization of functional monomers and a cross-linking agent in the presence of a molecule called a template. The result of the synthesis is a polymer that specifically and selectively captures the standard or its structural analogs. MIPs are chemically and thermally stable and resistant to various physicochemical factors (e.g. high pressure, various types of radiation). Due to their properties, polymers with molecular footprints are used, among others, in analytical methods (juko-sorbents in solid phase extraction, sensors, column fillings), as drug release systems, catalysts in synthesis. In my work I deal with simulations of prepolymerization systems, polymers, the adsorption process and theoretical analysis of interactions determining molecular recognition in MIPs systems. I also conduct experimental work involving the synthesis and research on the properties of MIPs. Despite the favorable properties of traditional MIPs (obtained by block or precipitation polymerization), there are limitations in their use related to the time-consuming separation of the appropriate bed or the pattern remaining in the polymer matrix after the treatment process. One way to overcome these difficulties may be to obtain magnetic molecularly imprinted polymers (MMIPs). Their isolation from the reaction mixture does not require centrifugation and filtration and is easier than with traditional polymers due to the possibility of using an external magnetic field. The external magnetic field simplifies the application of MMIPs when separating compounds and as drug carriers to a target site in the body. This project covers preliminary studies on the preparation and analysis of the properties of MMIPs. As part of the project, I am going to obtain MMIPs intended for the separation of biogenic amines. In the first stage, the synthesis of iron (II) oxide (III) (Fe3O4) will be performed. The next step will be to modify the surface of magnetic particles (magnetic nanoparticles, MNPs). For this purpose, the MNPs will be coated with a SiO2 layer using tetraethoxysiloxane and then functionalized with a suitable silane and optionally a surfactant. In the next stage, the synthesis of appropriate polymers will be performed with the molecular trace of the selected pattern. As a reference, I will use NN-dimethyl-2-phenylethylamine, which is a structural analog of the biogenic amine - 4- (2-dimethylaminoethyl) phenol hordenine. I am going to make four polymers with a molecular footprint using different functional monomers: acrylic acid, methacrylic acid, 4-vinylbenzoic acid and itaconic acid. The final process will be to remove the standard through Soxhlet extraction. The analytical part will include the analysis of the adsorption capacity and affinity of the obtained polymers to hordenine. In the analysis of adsorption properties, I will use the stationary method. I will choose one polymer for further research, showing the best properties (the highest affinity to the analyte and the highest adsorption capacity). I will determine the physicochemical parameters of the selected polymer, such as: adsorption kinetics, size and surface of the grains, chemical composition, magnetic properties, and I will perform a thermogravimetric analysis. I will optimize the adsorption process by examining the influence of pH, temperature and ionic strength of the solution on the adsorption capacity of the analyte on the imprinted polymer. I will examine the selectivity of the polymer in relation to various biogenic compounds (tyramine, synephrine, octopamine, tryptamine, tyrosine) and the ability of the polymer to adsorb hordenine in model and biological samples. Hardenin was selected as the target analyte because, to the best of my knowledge, there are no literature reports on molecularly printed polymers for the selective adsorption of this biogenic amine. At the same time, hordenine affects the functioning of the body: it stimulates the release of gastrin, inhibits monoamine oxidase B, and also inhibits melanogenesis in human melanocytes. Due to the possibility of its consumption in fruits, herbs and dietary supplements, there is a need to monitor its concentration in blood and urine. The most frequently described method of quantifying hordenine in the literature is HPLC-UV, which requires isolation of the compound and elimination of the matrix by solid phase extraction. Unfortunately, commercially available sorbents do not fulfill their role in the extraction of hordenine. Therefore, there is a need to find a selective sorbent that will allow a simple and effective extraction of the analyzed amine from complex samples, while eliminating the influence of the biological matrix on further quantitative determinations. The expected effect of the activities described in the project will be to increase the knowledge of MMIPs, interfering compounds and the process of molecular imprinting of amines. The obtained MMIP will facilitate the quantification of hordenine in biological samples. The obtained results of preliminary studies will help to develop optimal conditions for obtaining and analyzing MMIPs intended for the separation of various analytes and will be helpful in further work on the preparation of drug forms enabling the release of the active substance in a specific place or organ in the organism.

My research interests include the pathogenesis of atherosclerosis, I focus mainly on determining its molecular mechanisms. The main goal of my research is to develop effective preventive and therapeutic methods. My research to date has focused on assessing the impact of pro-atherogenic factors on dysfunction; vascular endothelium. They include assessments of the role of oxidative stress in activating the inflammatory process in endothelial cells by inducing the expression of adhesion molecules (ICAM, VCAM), chemotactic factors (MCP-1), and activation of mitogen-activated kinase signaling pathways (ERK, JNK, p38) and the nuclear factor - KB The aim of the research is to evaluate the expression and function of innate immunity receptors in the development of inflammation in endothelial cells. The research will be carried out using: - cell cultures - endothelial cells from umbilical veins (HUVEC), - atherosclerotic plaque preparations (N 40), material currently collected from patients from the carotid artery during elective open endarectomy - the research will be performed in cooperation with the Department of Otorhinolaryngology of the Faculty of Dental Medicine of the Medical University of Warsaw (consent of the bioethical committee KB / 92/2008) - we have the clinical characteristics of these patients.  The implementation of the research includes: - determination of the expression level of the following receptors: TLR 2, TLR 4, TLR 7, TLR 9, RAGE, as well as the H1MGB1 protein and nuclear factor - KB in umbilical vein endothelial cells (HUVEC) before and after stimulation with ligands of the tested receptors by cytometry flow cytometry - evaluation of the effect of inhibitors of individual receptors on ligand-stimulated expression of the KB nuclear factor, HMGB1 protein (flow cytometry) and expression of IL-6 in endothelial cells (Elisa test) - evaluation of the expression of TLR 2, TLR 4, TLR 7, TLR 9, RAGE receptors and proteins H1MGB1 and nuclear factor - KB in atherosclerotic plaque by immunohistochemistry. Available scientific data indicate an important role of innate immunity receptors, ie: toll-like receptors (TLR) and receptors for advanced glycation end products (RAGE) in the pathogenesis of atherosclerosis. Improper activation of these receptors causes disturbances in the body's homeostasis. The lack of signaling by these receptors exposes the body to pathogenic attack, while over-activation causes the uncontrolled release of many pro-inflammatory cytokines and chemokines, leading to the development of inflammation, which in turn intensifies the development of the atherosclerotic process. The main challenge for the future of successful targeted therapy in patients with atherosclerosis is to inhibit the inflammatory process associated with this disease without affecting the body's innate immunity. Therefore, it is important to understand the signal transduction pathways and the function of individual innate immunity receptors and the role of their selective inhibitors. This will control the immune response and block negative processes for the host. Currently, we have only selective data in this area and it is necessary to conduct research documenting the impact of inhibition of the activity of these receptors on the development of atherosclerosis. The planned research will provide important and missing information on the role of innate immunity receptors in the induction of inflammation in endothelial cells. The effect of selective inhibitors of these receptors will be documented, the effect of which will indicate the possibility of using these inhibitors in the treatment of inflammation, which will allow the control of the immune response and blocking inflammatory processes without affecting the organism's natural protective barrier. Assessment of the severity of inflammation by determining the expression of selected receptors in atherosclerotic plaque collected from patients from the carotid artery will provide new data on the role of these receptors in regulating inflammatory processes. The conducted research will be the basis for the development of new, effective therapeutic strategies in the prevention and treatment of atherosclerosis.