Ministry of Science and Higher Education

Circulating microRNAs (miRNAs) secreted mostly by platelets can significantly affect the concentration of miRNA in plasma, which makes it possible to monitor platelet function in populations of patients with increased cardiovascular risk, including type 2 diabetes. The assessment of platelet reactivity has many limitations, which means that the prognostic value of the phenomenon of increased platelet activity thus assessed remains uncertain. To date, no studies have investigated the potential relationship between the profile of circulating miRNAs and the prognosis in patients with type 2 diabetes.

Recent studies indicate that the pathogenesis of arterial hypertension may be related to disturbances in the composition of the intestinal flora. The commensal colonizing bacteria in the gut metabolize nutrients to produce a range of compounds that enter the bloodstream. The aim of the proposed project is to study the role of indole, a metabolite of intestinal bacteria, in the regulation of blood pressure and the development of hypertension. Haemodynamic studies will be performed in normotensive animals (WKY rats), hypertensive animals (SHR rats) and rats with borderline arterial pressure (BHR rats). The effect of increased indole levels in the large intestine, peripheral blood and cerebrospinal fluid on the regulation of blood pressure and the development of hypertension will be investigated. In addition, possible mechanisms by which indole could act in the body will be explored. Hypertension and its complications are one of the main causes of morbidity and mortality in Poland and the EU. The results of the proposed study may contribute to understanding the role of indole, a metabolite of the intestinal flora, in the regulation of blood pressure and in the pathogenesis of hypertension. The study will assess the therapeutic potential of indole synthesis donors / inhibitors in cardiovascular diseases, especially in arterial hypertension.

Recurrent nodal tachycardia (AVNRT) is one of the most common supraventricular arrhythmias (SVT) in the population and one of the most frequently ablated (approximately 1: 1,000 in the general population). Despite such a high frequency of AVNRT, only a few reports of a familial AVNRT-FAVNRT (familial AVNRT) occurrence have been published so far. The authors of these publications indicated the presence of FAVNRT most often in two family members with first-degree kinship. So far, data have been published in the form of original articles and abstract reports on a total of 41 families worldwide with confirmed FAVNRT. The population of FAVNRT patients from the Podkarpacie region presented by S. Stec and co-authors was the most numerous and accounted for up to 15% of FAVNRT among subsequent patients qualified for ablation procedures due to AVNRT. These observations prove the existence of a unique and the most numerous in the world population of FAVNRT families in the Podkarpacie region, and the sketched pedigrees and probability calculus prove the existence of a genetic basis for the occurrence of FAVNRT in this population. Research hypothesis: 1. Genetic factors are responsible for the occurrence of AVNRT in individuals with first or second degree relationships. 2. The frequency of AVNRT among family members with FAVNRT is higher than the frequency of AVNRT in the general population. Objectives of the study: 1. Keeping a registry of patients with FAVNRT occurring in the Podkarpacie region in order to accurately assess the clinical characteristics and occurrence of FAVNRT in this region of Poland. 2. Establishing the genetic basis for the occurrence of FAVNRT in the population of AVNRT patients for the first time in the world. 3. Determination of the influence of the genetic basis of FAVNRT on the mechanisms of AVNRT formation and the properties of the atrioventricular junction in the conduction of other cardiac arrhythmias.

Scientific objective: The aim of the project will be to evaluate the type and course of cardiomyocyte death in the female rat Takotsubo (TTC) model of isoprenaline-induced cardiomyopathy (ISO). State of knowledge: Takotsubo cardiomyopathy (TTC) is characterized by acute, reversible systolic left ventricular failure caused by emotional and / or physical stress, mainly in postmenopausal women. TTC shows akinesia of the apical segments of the left ventricle (echocardiography) and its specific systolic shape (ventriculography). The pathogenesis of Takotsubo cardiomyopathy has not yet been elucidated. It has been suggested that it may result from coronary circulatory disorders or the cardiotoxic effect of catecholamines. Recently, a male rat model of TTC induced by intraperitoneal administration of isoprenaline, an agonist of P1- and P2-adrenergic receptors, has been developed. Recently, our research team developed the first female rat TTC model induced by IP injection of ISO. Few studies, including our preliminary published results of TTC histopathological changes in the heart, indicate the presence of cardiomyocyte death and the accompanying inflammatory response. The type of cardiomyocyte death in Takotsubo cardiomyopathy, its impact on the course of the disease and the role of female sex hormones in these processes have not yet been elucidated. It seems that three types of cardiomyocyte death in particular may occur in the course of TTC, ie apoptosis, necroptosis and pyroptosis. Apoptosis is programmed cell death that is dependent on caspase 3. Necroptosis combines the features of programmed apoptosis and necrosis. Activation of the necroptotic pathway requires recruitment of the RIPK1 (Receptor-Interacting Protein Kinase 1) and / or RIPK3 (Receptor-Interacting Protein Kinase 3) and MLKL (Mixed Lineage Kinase domain -Like) proteins. Necroptosis has also been shown to play a significant role in inducing an inflammatory response. Pyroptosis, on the other hand, is a pro-inflammatory, programmed type of cell death, in which the key stage is the recruitment of the cytosolic protein complex called the inflammasome, e.g. NLRP3, activating caspases 1 and 11, inducing the release of pro-inflammatory interleukins (IL): IL-1P and IL-18. There are data suggesting the involvement of NLRP3 in the pathogenesis of numerous diseases of the cardiovascular system. Type II calcium-calmodulin-dependent kinase (CaMKII) is a protein directly involved in cardiomyocyte apoptosis, induced by stimulation of P1-adrenergic receptors and necroptosis of these cells associated with inflammation. The expression of this kinase increases with low estrogen levels, suggesting that it may have a significant influence on the pathogenesis of TTC. Methods and description of the research: The research will be carried out in 160, 9-week-old female Sprague Dawley (SPRD) rats. The animals will be divided into 4 treatment groups (5 runs in each group, 8 animals in the series). At 9 weeks of age, rats will undergo bilateral ovariectomy (groups I and II) or sham surgery (groups III and IV). At 12 weeks of age, rats will receive an intraperitoneal injection of 150 mg / kg body weight. ISO (groups II and IV) or saline (NaCl) (groups I and III). In each of the study groups, 6, 12, 24, 72 hours and 10 days after the ISO or NaCl injections, transthoracic echocardiography will be performed and blood and heart will be collected for biochemical tests. The plasma concentration of caspase 1, 3, IL-1P, IL-18, RIPK1, RIPK3, MLKL and CaMKIIS (ELISA) will be determined. Hearts will be tested for caspase 1, 3, RIPK1, RIPK3, MLKL and CaMKIIS (Immunohistochemistry). Characterization of the final result: So far, the process of apoptosis, pyroptosis and necroptosis in TTC has not been characterized, and their influence on the course of the disease has not been explained. The present study will bring closer the role of the cardiomyocyte death process in TTC and the influence of female sex hormones on the occurrence of the above types of cell death, which in the future will be the starting point for further research to determine an effective targeted therapy. Assessment of the plasma concentration of proteins involved in apoptosis, necroptosis and pyroptosis may contribute to the detection of new prognostic markers in TTC that are useful in clinical practice.

Regulatory T cells (Treg) are responsible for suppressing the immune system response. They play a key role in the proper functioning of the immune system. Their abnormal function is seen in many diseases, such as autoimmune diseases, allergies, as well as in the process of transplant rejection. The search for compounds that induce these lymphocytes or enhance their activity is of great importance in the development of new methods of treating many diseases. The proposed studies will determine the effect of various immunomodulators and drugs on the activity of T regulatory lymphocytes. Lymphocytes will be isolated from the peripheral blood of healthy donors (coats), and then Treg lymphocytes (lymphocytes suppressing the immune response) will be induced in the culture. In the preliminary studies to date, we have observed that butyric acid and inosine pranobex increase the number of Treg lymphocytes (CD4 + CD25h19hFoxp3 + phenotype) in culture. In addition, therapeutics such as rapamycin, glatiramer acetate, atorvastatin, vitamin D3 and prednisolone will be investigated. In the next stage, it will be shown whether the tested compounds affect the functionality of Treg lymphocytes. For this purpose, a functional test will be used to determine how lymphocyte function changes depending on the drug used. This test will determine whether these therapeutics could potentially be used to stimulate the activity of pathological regulatory T lymphocytes in patients with their deficiency. It will consist in testing the ability of Treg lymphocytes, after their incubation with the investigational drugs, to inhibit the proliferation of effector lymphocytes. In the last stage of the experiment, it will be examined whether the tested drugs can be used for therapeutic purposes in patients suffering from myasthenia gravis - a representative autoimmune disease. Treg lymphocytes in patients from this group have impaired function, which is one of the reasons for the onset of the disease. Lymphocytes from the peripheral blood of patients with newly diagnosed myasthenia gravis will be isolated, and then Treg lymphocytes will be induced in in vitro culture. It will then be determined whether the immunomodulators restore the normal activity of the pathological lymphocytes in the functional test described above using peptides derived from acetylcholine receptors (AChR) as stimulators of specific reactions. The proposed research may be important in the treatment of, inter alia, autoimmune diseases, allergies, graft versus host disease, or in inducing transplant tolerance.

Non-Hodgkin's lymphomas account for about 2% of all malignant neoplasms in Poland, and their incidence has been systematically increasing in recent decades. Despite the wide availability of various forms of treatment, patients' response to therapy is not satisfactory. A significant problem is the resistance of cancer cells to the drugs used. The PI3 / AKT pathway plays a key role in the development of resistance to chemotherapy. Literature data show that some of the chemotherapeutic agents used in lymphoproliferative diseases can activate AKT kin / e. The results of our research have shown that AK T kinase can, in turn, regulate the amount of the CD20 antigen - a target protein for therapeutic monoclonal antibodies also used in the treatment of lymphomas. Most of the treatment regimens currently used are based on combining chemotherapeutic agents with immunotherapy. Hence, the understanding of the interactions between these two groups of drugs is extremely important in the context of selecting the best possible combinations of these drugs. The scientific goal of the project is to investigate the influence of selected chemotherapeutic agents on the amount of CD20 antigen in cancer cells and on the effectiveness of therapeutic monoclonal antibodies directed against this molecule. We intend to carry out the experiments in models of established cell lines derived from B lymphocytes as well as on cells isolated from patients with hematological hyperplasia. The results of our research will make it possible to select chemotherapeutic agents, which could increase the effectiveness of aiUy-CD20 immunotherapy. This will make it possible to propose the most advantageous combination therapy regimens with the use of both groups of Icków [sic! - see Polish version]. In a broader perspective, our project will contribute to a better understanding of the mechanisms of secondary resistance to chemotherapeutic agents.