National Science Centre

The aim of the study will be to broaden the knowledge about oleacein, a compound that has unique properties and can be used in the prevention of both early and late atherosclerotic changes. The planned activities are of basic research nature. Assessment of cholesterol removal from foam cells by the SRB1 receptor and ABC Al transporter as well as ABCG1 transporter will at least partially explain the mechanism of action of oleacein in this area, which is still unresolved. Oleacein present in olive oil, olives and leaves of the European olive as well as the leaves of the native species - the common ligust, shows strong antioxidant and anti-inflammatory properties. Spectacular studies (US patent 9,682,056 B2) have shown that oleacein, by increasing the expression of macrophage scavenging receptors CD 163 and IL-10 and increasing OH-1 secretion, can reduce intracellular microbleeding and thus prevent plaque rupture [1]. Stabilization of atherosclerotic plaque by oleacein was confirmed in ex vivo studies carried out on atherosclerotic plaques collected from carotid arteries from patients following endarterectomy [2]. Current studies confirm that oleacein regulates the work of scavenger receptors (SRA, CD36, CD68 and LOX-1) responsible for cholesterol uptake by macrophage cells. Reduction of the expression in the above-mentioned receptors may lead to inhibition of foam cell formation, which results from the accumulation of oxidized LDL (oxLDL) by macrophages. Foam cells are the basic building block of the atherosclerotic plaque responsible for the development of atherosclerosis, leading to stroke and / or heart attack and death.

The main goal of this project is to obtain by chemical synthesis a series of new compounds from the bis-amidine group and preliminary analysis of their biological activity. This research will contribute to broadening the knowledge of organic synthesis (classical and using microwave radiation) used to obtain new chemical compounds. These studies will also allow the extension of the library of chemical compounds, which may be used in the future in various scientific research into other topics, but related to pentamidine derivatives. In my research work, I search for new chemotherapeutics in the group of compounds with two amidine groups, the so-called bis amidine. The best known drug in this group is pentamidine, used in the treatment of pneumocystis pneumonia (Pnemocystis carinii infections) and in the treatment of tropical diseases caused by protozoa (e.g. leishmaniasis). Compounds from the bis-amidine group are characterized by broad biological activity. Their activity may be e.g. antiprotozoal, antifungal, antimicrobial, antitumor, anti-inflammatory, they act on NMDA receptors. I have developed relatively efficient methods for the synthesis of new bis-amidines. They are obtained mainly by transforming the cyano group of the previously obtained bis-nitriles, which I synthesize with various methods, mainly in O-alkylation reactions of 4-hydroxybenzonitriles or aromatic nucleophilic substitution of para-substituted halogenobenzonitriles. I transform the cyano group mainly on the basis of proven methods: Pinera or obtaining an oxime and then reducing it. So far I have received several series of new bis-amidines with proven anti-Pneumocystis carinii and antibacterial activity. After receiving, appropriate purification and confirmation of their structure, the compounds are subjected to preliminary tests of biological activity. On the basis of these studies, I conduct a SAR (Structure Activity Relationship) analysis and select structures for further in vitro research and select leading structures for further syntheses. The project provides for obtaining a series of new compounds from the bis-amidine group with the expected high biological activity. I designed the compounds based on a literature review and with the support of molecular modeling methods. The structures of new compounds were also planned based on my previous research, in which I obtained a series of compounds with a number of different substituents on aromatic rings connected with each other by aliphatic or aliphatic-aromatic linkers. The activity profile (including their anti-P. carinii and antibacterial activity) was determined for the compounds obtained so far. The obtained results are optimistic and encourage further research. I have established, inter alia, that the introduction of nitro and amino groups into the rings significantly increases the activity of the compounds. These compounds act on P. carinii in nanomolar concentrations. I plan to obtain a series of new bis-amidines with asymmetric structure (differing in substitutions in aromatic rings) and a series of new symmetrical compounds with various combinations of substituents, the presence of which, according to the conducted SAR analysis, increases the activity of new derivatives. The compounds that I planned to synthesize under this project will be obtained as a result of multistage organic syntheses, in selected cases assisted by microwave radiation. Organic synthesis is based on certain and proven chemical reactions. All derivatives will be purified by means of crystallization and column chromatography (gravity or flash). The structure of new compounds will be confirmed by 1H NMR, 13C NMR, HSQC, HMBC, 13C CP / MAS, MS spectra, elemental analysis and crystallographic methods for selected compounds. Pneumocystosis, or severe opportunistic pneumocystic pneumonia (PCP - pneumocystis carinii pnemonia), affects people with a compromised immune system. This especially refers to AIDS patients. Although the number of PCP cases in the HIV-positive group has decreased with the introduction of HAART (High-Active Antiretroviral Therapy), PCP remains the main cause of mortality in this group. In addition, in recent years there have been many reports of PCP in patients after transplantation, chemotherapy for cancer treatment, chronic corticosteroid treatment or patients with autoimmune diseases (e.g. Crohn's disease). PCP is a disease that is very difficult to treat due to the unusual nature of the pathogen (insensitivity to anti-fungal drugs and most antibiotics) and relatively high toxicity of the drugs used (e.g. pentamidine). Therefore, there is a great need to search for new compounds with high biological activity that could potentially be used in the treatment of PCP in the future. 4. Expected effect. The main effect of the presented project will be the receipt, purification and confirmation of the structure of a series of planned compounds with the expected biological activity. The results of the work will be published in international journals. I plan to obtain a series of new bis-amidines of asymmetrical and symmetrical structure with various ring substituents. Such derivatives have not been described in the literature so far, nor have they been tested for their antibacterial and anti-Pneumocystis carinii activity. The project will enrich the library of pentamidine derivatives with new compounds. The obtained results will be part of the material for the project manager's habilitation dissertation. Due to the fact that this project is part of the medical chemistry framework, it will be necessary to conduct biological activity tests (apart from the presented application). The activity of the compounds will be determined on the basis of the in vitro activity study against Pneumocystis carinii (as part of scientific cooperation with the Department of Internal Medicine, University of Cincinnati) and antibacterial (as part of the scientific cooperation with the National Medicines Institute).

The pharmaceutical industry is one of the fastest growing and most profitable sectors of the global economy. Thanks to its existence to the possibility of treating an increasing number of diseases. This fact is also related to the need to develop modern science in the world, especially that related to the search for new chemotherapeutic agents. Neoplasms are the second cause of death after cardiovascular disease, not only in Poland, but also in Europe and the USA. Chemotherapy plays an extremely important role in the treatment of neoplastic diseases. A characteristic feature of many anti-cancer drugs is their limited effectiveness and extensive side effects that often make it impossible to continue the therapy. Therefore, the search for new anti-cancer drugs is one of the key directions of scientific research. The availability of modern technologies and the related development of research on the pathogenesis of cancer have become the basis for the development of various strategies of searching for new drugs. One of them is the search for natural substances that, in an unchanged or modified form, exhibit high antiproliferative activity against neoplastic cells and can be used alone or in combination with other drugs (combination therapy) in the treatment of neoplastic diseases. In the literature, there are more and more reports on new active substances that have a strong antibacterial, antifungal or anti-cancer effect, but nevertheless, the search for compounds that would show high efficiency in inhibiting the multiplication of neoplastic cells, while not adversely affecting the functioning of healthy cells, is still ongoing. It is worth noting that there are more difficulties than one might expect. The complicated chemical synthesis itself poses many problems, be it with the preparation of the substance, its modification or with its purification in order to obtain a product of high purity. Another problem is the solubility of this substance. Chemical compounds have different chemical and spatial structures, which affects their behavior in solvents, including body fluids. It must be remembered that the undissolved compound will not be able to penetrate into the cells and cause the desired effect. Therefore, the challenge is not only to invent an appropriate compound, its chemical synthesis (or modification of an existing substance of natural origin) but also to enable the product to be well soluble. Many organic substances are easily dissolved in organic solvents, while the body fluids in which the substances will act contain water, which unfortunately does not dissolve organic molecules. The aim of this project is to search for new derivatives of plant-derived substances (alloxanthoxylethins), which will show antiproliferative activity against cancer cells comparable to that of leading cytostatics, and reduced cytotoxicity against normal cells. Commonly known substances characterized by high lipophilicity are fatty acids, which have a high affinity for biological membranes and can influence their structure and, consequently, the properties of the membrane, its permeability and selectivity. Planned modifications by combining alloxanthoxyletine with selected fatty acids will change the lipophilic properties, which will result in an increase in the cytostatic activity of these substances.

Chronic otitis media with cholesteatoma is still a major therapeutic problem. During the development of cholesteatoma, the cells of the keratinized epithelium from the external auditory canal to the middle ear cavity "crawl" and proliferate, causing chronic inflammation as a result of the activation of the immune system. As a result, the structures of the middle and inner ear are destroyed, leading to hearing loss, paralysis of the facial nerve, balance disorders (vestibular dysfunction), as well as a risk of intra-temporal and intracranial complications.The reasons for the formation and development of cholesteatoma are not fully understood. Surgical treatment is the treatment of choice. In our previous studies, we have shown a significant role of innate immunity in the development of acquired middle ear cholesteatoma. Our preliminary studies showed the presence of the HMGB1 protein and its RAGE receptor, as well as the presence of Toll-like receptors in the cholesteatoma microenvironment. Currently, more and more data indicate the key role of extracellular vesicles (EV) in the pathogenesis of chronic inflammation, because EVs act as carriers of various proteins, including pro-inflammatory modulating the activities of other cells. Taking into account the results of our preliminary studies, we hypothesize that by releasing HMGB1-containing microvesicles from the microenvironment of cholesteatoma, which is also spread systemically through the circulatory system, local and general inflammation occurs. Due to the fact that microvesicles can be collected by endocytosis by cells of the immune system, such as macrophages, but also by other cells (fibroblasts, vascular endothelium), we want to assess the direct impact of microvesicle HMGB1 (EV-HMGB1) on these cells in research in vitro and ex vivo. We also assume that the assessment of the number of microvesicles and the level of HMGB1 in the serum may be a marker of inflammation and correlate with the clinical advancement of cholesteatoma or its recurrence.

Aim of the project: The aim of the project is to evaluate the influence of iron and zinc on NETs ejection. In addition, by examining net ejection in patients with iron deficiency anemia and haemochromatosis (a disease characterized by the accumulation of iron in the body), I will assess whether the deficiency and excess of the test metal affects the ability of neutrophils to release the net and the efficiency of killing pathogens within the NETs. For the last stage of the research, I will include mice with haemochromatosis and mice fed with poor fodder and saturated with zinc and iron. Based on the results of research in a mouse model, I will try to explain whether a diet low in micronutrients or excessive supplementation of these elements may adversely affect the mechanisms of innate immunity. Expected results: The results of our research will contribute to a better understanding of how homeostasis of selected metals influences granulocyte function, including NET ejection. More importantly, we will estimate the effect of excessive micronutrient supplementation on non-specific immunity.

The general aim of this study is to deepen the knowledge of the experience of physical pain in people addicted to alcohol, a comprehensive analysis of pain sensitivity and endurance in this group, and to assess the relationship between these parameters and other risk and prognostic factors in alcohol dependence described in the literature. Detailed research goals:1) Analysis of the relationship between pain sensitivity and the known risk factors of relapse to drinking in people addicted to alcohol, with particular emphasis on the assessment of decision-making and processing of emotions in the study group. 2) Analysis of the relationship between pain tolerance and known risk factors of relapse to drinking in people addicted to alcohol, with particular emphasis on the assessment of decision-making and emotional processing in the study group. 3) Comparative analysis of subjective and objective measures of tolerance and pain sensitivity in a group of alcohol addicts. EXPECTED RESULTS: We hypothesize that people addicted to alcohol are characterized (compared to non-addicts) by a lower tolerance of pain and a greater sensitivity to pain. We anticipate that the experience of physical pain will be significantly associated with recognized risk factors for addiction development and relapse to drinking: depression, poorer behavioral control, sleep disturbance, history of suicide attempts, and poorer emotional regulation. Increasing the knowledge of the relationship between the various dimensions and characteristics of pain and other known risk factors for relapse drinking (impulsivity, emotional intelligence, emotional regulation, sleep disturbances, depressive symptoms) could have significant cognitive and clinical significance. It could also provide arguments to verify in a clinical setting the concept of addiction as "chronic emotional pain syndrome". In our opinion, this, in turn, may potentially contribute to the modification and improvement of the still very unsatisfactory results of treatment of people addicted to alcohol.

Project goals. Mesenchymal stromal cells (MSCs) have the ability to influence the activity of various types of immune cells and are currently being tested as a therapeutic agent in numerous clinical trials. For several years, attention has been drawn to the fact that hypoxic conditions significantly affect the immune response profile. It is also known that the state of reduced oxygen availability in the environment stimulates in MSC the activation of HIF-1, a transcription factor that influences the expression of several hundred genes. However, the importance of HIF-1 activation for the interaction between immune cells and MSCs has not yet been thoroughly investigated. The aim of this project is to determine the influence of HIF-1 on the immunomodulatory properties of MSC cells. The main research hypotheses of this project are the following: 1) HIF-1 activation significantly influences the immunomodulatory effect of MSC; 2) by modifying the activity of HIF-1 in MSCs using chemical methods or cellular engineering, and also in combination with inhibitors of selected cytokines (e.g. IL-6), the therapeutic potential of MSCs in terms of immunomodulation can be influenced; 3) the effect of HIF-1 activation on the immunomodulatory properties of MSCs varies depending on the origin of the cells. Expected results. The immunomodulatory properties of MSCs raise great hopes. However, the results of clinical trials to date are not as promising as the results of in vitro studies. Therefore, it is crucial to conduct further basic research using modern analytical methods that will deepen the current knowledge about the potential of MSC. The determination of the impact of HIF-1 activation in MSCs on their immunomodulatory properties is intended to allow for a better understanding of the mechanisms of the physiological and therapeutic effects of MSCs.We expect that determining the effect of hypoxia on the interactions between MSCs and inflammatory cells will provide a better understanding of the differences in the effectiveness of using MSCs in various disease states. Finally, the obtained results can form the basis for the definition of new strategies for enhancing the immunomodulatory potential of MSCs. Units involved: The project manager is Prof. Leszek Pączek. The research will be performed mainly in the cell culture laboratory of the Department of Immunology, Transplantology and Internal Diseases of the Medical University of Warsaw under the direct supervision of Dr Anna Burdzińska. The second unit involved in the implementation of the project will be the Department of Clinical Immunology of the Medical University of Warsaw under the supervision of dr hab. Radosław Zagożdżon. Human cells will be obtained from the bone marrow (cooperation with the Department of Orthopedics and Traumatology of the Motor System of the Medical University of Warsaw), and also from umbilical cords (cooperation with the Department of Obstetrics and Gynecology of the Medical University of Warsaw).

The aim of the project is to try to combine these two novel therapeutic concepts by evaluating the efficacy of the novel CAR receptor targeting PD-LI on cancer cells or tumor stromal. For this purpose, the following research tasks are planned: Optimization of CAR-anly-PD-LI receptor expression methods in effector cells. Comparison of the anti-tumor efficacy of CAR-T and CAR-NK cells against tumor cells showing different levels of PD-LI expression. Use the synNotch strategy to achieve specific CAR-anly-PD-LI action in the vicinity of tumor cells. Evaluation of the efficacy of cytotoxic cells expressing CAR-anti-PD-Ll receptor in vivo. Expected results: At the present time, intensive research is being carried out into the use of receptor technology to fight as many types of cancer as possible. The focus was on, inter alia, searching for new therapeutic targets; however, it is becoming increasingly clear that cancer cell resistance is a significant problem in this therapy. The project has the potential to make significant progress on both of these aspects by providing conceptual evidence for the effective and safe targeting of effector cells to a therapeutic target, extremely important in tumor resistance to immunotherapy. The research results will contribute in the future to the optimization of therapeutic regimens used in oncology with the use of modern molecularly targeted immunotherapies.

The aim of the project is the aim of the research project is to analyze the neurobiological mechanisms related to the control of behavior and the recognition of emotional states by people diagnosed with pedophilia. It is planned to include the perpetrators of pedophilic acts as well as people who did not come into physical contact with the child. The specific objectives of the study are: Identification of neural networks in the central nervous system (CNS) that are related to the recognition of emotional states in adults and children (Reading Mind in the Eyes Test test, RMET) and to assess whether the processes of recognizing emotions are related to the type of sexual intercourse preferences in the surveyed people. Assessment of the relationship between the type of sexual preferences and the activity of the CNS during tests assessing the ability to inhibit the reaction (Stop-Signal Task, Affective Go/No-go test). Analysis of the neurobiological correlates of the cognitive processes involved in performing the Sexual Preference Implicit Association Test. We hypothesize that during the performance of the above-mentioned tests, the activity of the central nervous system in people diagnosed with pedophilia will be significantly different from the activity of the CNS observed in people from the control groups. The expected results, comparing the four groups selected for different sexual preferences and the ability to control one's own behavior and recognize emotional states, may constitute a significant step forward in understanding complex mechanisms of behavior control and emotional processing. Abnormalities of the above-mentioned processes play an important role in many mental disorders, however, a comprehensive description of these phenomena may also have an extremely valuable cognitive meaning for the entire population. The results of this project may lay the foundations for a novel model of the neurobiological determinants of pedophilia, which could become the basis for inspiring theoretical concepts regarding the etiology of sexual crimes. These results may also help to create new therapeutic programs for perpetrators of pedophilic acts, which seems to be extremely important in the context of the results of meta-analyzes indicating the surprisingly low effectiveness of the currently proposed interventions.

The aim of the project is to investigate the project to investigate the role of reactive nitrogen species in the formation of NETs, ​​including the mechanism underlying the formation of RFA-induced NETs, ​​and to check whether their participation is necessary for this process. Expected results The formation of extracellular neutrophilic networks has been discovered relatively recently, and although this process has been the subject of much scientific research, our knowledge of the regulation and course of this phenomenon remains incomplete. The research carried out in this project will provide a better understanding of the mechanisms of non-specific immune responses as well as our understanding of the modulating role of RFA in the functioning of cells of the immune system. It is worth noting, however, that detailed research on the factors responsible for the regulation and course of the phenomenon of networking is interesting not only because of the progress in science. In the future, they could have an impact on clinical medicine, pointing to new treatment strategies for many inflammatory and autoimmune diseases where excessive networking is involved in pathogenesis.