National Science Centre
Endometriosis is a common, chronic gynecological disease, manifested by the presence of endomeric tissue outside the uterus, most often in the smaller pelvis. The disease is accompanied by chronic pain and it is one of the main causes of infertility, which makes it a significant clinical and social problem. The etiology of endometriosis is unclear, and the underlying pathological mechanisms are still poorly understood. The presence of endometriosis foci is most likely the cause of the induction of chronic inflammatory reactions manifested by the accumulation of activated macrophages and lymphocytes and the secretion of large amounts of various pro-inflammatory cytokines. Endometriosis is also associated with autoimmune disorders and therefore some consider it an autoimmune disease. It has been suggested that endometriosis and the associated immunopathogenetic phenomena may be caused by a disturbance in the distribution and activation of the Treg and Th17 lymphocyte systems. Treg cells perform suppressor functions, are responsible for the phenomenon of auto-tolerance and inhibit inflammatory and autoimmune reactions, while Th17 cells are responsible for stimulating immune responses, especially inflammatory and autoimmune responses. Our previous observations show that endometriosis is associated with an increase in the number of Treg cells in the peritoneal fluid, which may be important for the course of the immune phenomena underlying this disease. MHowever, the mechanism responsible for this phenomenon remains unknown. The role that Th17 cells may play in endometriosis is also unknown. We hypothesize that the disturbance of Treg cell distribution and, possibly, Th17 cells is associated with chemotaxis and differentiation of these cells under the influence of the cytokine-rich peritoneal fluid environment. Therefore, the aim of our project is to investigate the influence of the peritoneal fluid environment of patients with endometriosis on chemotaxis and differentiation of Treg and Th17 cells, and to determine if there is a correlation between the level of specific cytokines / chemokines in the peritoneal fluid and the occurrence, migration and differentiation of these cells. The research will be conducted in close cooperation with the 1st Department of Obstetrics and Gynecology at the Medical University of Warsaw. The material used for the research will be the peritoneal fluid obtained during the laparoscopic procedure and peripheral blood from about 30 patients with endometriosis and a similar number of patients qualified for the control group. In the peritoneal fluid, the concentration of selected cytokines / chemokines will be determined by ELISA tests. The blood and peritoneal fluid subpopulations of Treg and Th17 will be determined by flow cytometry. The chemotactic and stimulating activity of the peritoneal fluid will be determined by in vitro assays using isolated Treg cells and a CD4 + cell population containing Treg and Th17 precursor cells. These cells will be isolated from buffy coat by immunomagnetic or flow cytometer (FACS Aria) sorting. Individual patient test results will be analyzed using multiple linear regression and logistic regression models. We believe that our proposed research will contribute to deepening the knowledge about the causes and mechanisms of Treg and Th17 cell system disorders in endometriosis. This knowledge may be useful in creating the concept of new methods of therapy for this frequent, but still mysterious, disease,
The aim of the project is to investigate the effect of selected kinase inhibitors (such as Syk, Btk, Akt, PI3K, mTOR) on the anti-tumor efficacy of monoclonal antibodies related to the mechanism of antibody dependent cellular cytotoxicity (ADCC). There are currently many clinical trials in which inhibitors of the cell signaling pathway are tested in combination with monoclonal antibodies used in cancer therapy. The results of our preliminary studies indicate that some of the tested inhibitors of kinases involved in signal transduction from the B lymphocyte receptor almost completely inhibit the cytotoxic functions of NK cells (which are the main effector cells in the ADCC process), and thus significantly weaken the anti-cancer effect of monoclonal antibodies in ADCC mechanism. As part of the project, the scope of research will be extended. Experiments will be carried out to test a series of various inhibitors of the aforementioned kinases on the activity of NK cells in the ADCC process induced by many different monoclonal antibodies (such as trastuzumab, obinutuzumab, alemtuzumab, cetuximab or daratumumab). The project will develop and optimize methods of studying the mechanisms of NK cell cytotoxicity, in particular the ability to kill target cells by the mechanism of cell cytotoxicity dependent on monoclonal antibodies commonly used in cancer therapy: anti-CD20 antibodies (rituximab, ofatumumab, obinutuzumab), anti-HER2 (trastuzumab), anti-EGFR (cetuximab) as well as anti-CD38 (daratuumab) used in clinical trials. In the ADCC assay models, the target cells will be cells of various human cell lines expressing the antigens recognized by the respective monoclonal antibodies: CD20 antigen-expressing lymphomas (Raji, Daudi, DoHH2, Ly-4), HER2/Neu expressing breast cancer lines (SKBR3 , T47D, ZR75-30) and ovarian cancer (SK-OV-3), showing EGFR expression: lung cancer (SW-900), bowel cancer (HCT116, LoVo), prostate cancer (Du-145). In addition, a method for studying NK cell degranulation and cytokine secretion using flow cytometry will be developed and optimized. Screening will then be performed using established methods to identify inhibitors of the cell signaling pathway that may affect the anti-tumor activity of NK cells. Inhibition of selected signaling pathways in NK cells will be confirmed by Western blotting. The project also plans to introduce a sequence encoding the FcγRIIIa (CD16) receptor, which is a key receptor involved in ADCC, into an NK-derived cell line by viral transduction. All commercial NK cell lines lack the surface of the FcγRIIIa receptor. The line created will serve as a model for ADCC research. Monoclonal antibodies are widely used in the treatment of cancer. There are currently 15 monoclonal antibodies registered in oncology. Moreover, new monoclonal antibodies modified in such a way as to maximize their antitumor effectiveness are still being tested. In the light of ongoing clinical trials combining monoclonal antibodies with many kinase inhibitors, it is extremely important to investigate how the tested chemical compounds affect the mechanisms of action of monoclonal antibodies already used in therapy. Understanding the interactions between the tested drugs will allow the selection of those therapeutic combinations that should be contraindicated, as well as those that should be used in combination therapy. The project's research will also contribute to a better understanding of NK cell biology and a more detailed understanding of what role different signal transduction pathways play in NK cell activation.
Photodynamic therapy (PDT) is an example of a method of cancer treatment in which activation of the immune system is one of the key elements responsible for the treatment effectiveness. By activating the photosensitizer and creating reactive oxygen species, PDT causes the breakdown of cancer cells. In this way, inflammation within the tumor is induced, and the activated elements of the non-specific immune response are able to initiate an effective specific response only in special cases. Due to the immune tolerance, despite the presence of cancer antigens, the immune system does not fight the disease effectively. The enzyme indoleamine 2.3-dioxygenase (IDO) inhibits the immune response both within the neoplastic lesion and promotes the development of immune tolerance to neoplastic antigens. IDO is synthesized by cancer cells, but its main source in the body are dendritic cells and macrophages. The enzyme inhibits the proliferation and activation of cytotoxic T cells and causes the differentiation of CD4 + T cells into regulatory cells. The aim of the project is therefore to investigate whether inhibition of IDO activity will affect the long-term effect of PDT. The research hypothesis assumes that abolition of the immunomodulatory effect of IDO will result in the destruction of both the tumor and secondary neoplastic lesions.
Breast cancer is the most common malignant neoplasm in women. Seventy percent of human breast cancers have the estrogen receptor, which is an extremely attractive target for endocrine therapies. However, a significant percentage of breast cancers treated this way become refractory to this therapy, which is associated with a significant worsening of the prognosis. This often occurs in the mechanism of switching the dependence of tumor growth from estrogen dependent on estrogen in the direction dependent on pathways activated by growth factors, their receptors (GFR) or GFR signaling oncogenic proteins. One of the factors responsible for this phenomenon seems to be the conditions of oxidative stress in neoplastic cells. It is the subject of intense research. Our group, coordinating the work of several centers abroad, recently found that one of the main chains of enzymes (dependent on thiol groups) counteracting oxidative stress protects the estrogen receptor against the effects of this stress. It has also been found in the clinical material from a total of over 1,200 patients that the increased expression in the tumor of the main enzyme of this chain, peroxyredoxin 1 (PRDX1), is a favorable prognostic factor. Based on our preliminary studies and the available results of other research groups, we have created a systemic model in which the enzyme chain PRDX1 / 2thyoredoxin (TXN) plays an important role in maintaining the estrogen / ER pathway dependent type of breast cancer growth. This hypothesis is the basis for setting three research goals to be carried out in vitro in this project. The fourth objective concerns in vivo research, complementing the holistic approach to the studied phenomenon. The four main goals of the current project are: Goal 1: To investigate the molecular mechanisms of ER protein suppression induced by a defect in the function of the thiol-dependent antioxidant enzyme chain. Objective 2: To investigate the role of thiol-dependent antioxidant enzymes in switching signaling in breast cancer between hormone-dependent and oncogen-dependent breast cancer using -omics techniques. Objective 3: To evaluate the regulation of PRDX1 / 2 enzymatic activity in breast cancer. Aim 4. To study the influence of PRDX1 dysfunction on the progression of breast cancer in the in vivo xenotransplantation model.
The aim of the project is to search for genetic variants predisposing to early death in the group of adults representative of the Polish population. The aim of the project is to answer the following questions: 1) What known genetic variants predisposing to early death are particularly common in the Polish population. What are the relative frequencies of each variant, and are there any variants that are currently underestimated in importance. 2) What new genetic variants (unknown mutations in genes with known function and mutations in genes not yet associated with diseases) occur in the Polish population, contributing to the early death of adults. Project results are expected to identify the most common genetic diseases contributing to early death among adults in Poland. With a high degree of probability, you can expect to find min. 10-20 cases of mutations with a clear effect and a clear relationship with the cause of death (previously described mutations, new destructive mutations in known genes). In addition, it is possible to identify new genes, the defects of which may cause serious, because they lead to early death, diseases in humans. Due to the representativeness of the WOBASZ cohort for the entire population of Poland, the results will allow to estimate the role of known rare diseases in adult mortality in our country. The results of the project may indicate the desirability of performing more extensive genetic tests for certain mutations in people with certain diseases than at present, or even the validity of screening the entire population. The emergence of new genes related to mortality is important for basic research in biology, physiology and medicine. The project is the first attempt to use WES in an epidemiological study in Poland. WES has been widely used in epidemiological studies in other countries, however, there are no reports on the use of this technology in the analysis of early mortality comparable to that described in the application.
Ovarian cancer has the highest mortality rate of any cancer of the female reproductive organs, with a 5-year survival rate of only 30% of patients. The reason for such a low survival rate of patients is late diagnosis, caused by the lack of clinical symptoms and the lack of appropriate biomarkers to detect the disease at an early stage. Importantly, while some of the ovarian cancer subtypes initially respond well to therapeutic approaches, subsequent resistance remains a significant clinical problem. Therefore, further research is needed to deepen the understanding of the molecular phenomena occurring in ovarian cancer cells, especially in the context of searching for new therapeutic strategies. As with other cancers, ovarian cancer uses a wide variety of mechanisms to suppress the activity of the immune system. The aim of the project is to comprehensively assess the role of two exosome-related enzymes (arginase 1 and 2) involved in the breakdown of endogenous amino acids in ovarian cancer's ability to escape immune surveillance. The development of chronic inflammation and the induction of enzymatic pathways for arginine breakdown are recognized as important factors involved in this mechanism. Thus, we hypothesize that by releasing arginase-containing exosomes which then spread systemically through the circulatory system, cancer cells lead to a global deficiency of L-arginine and thus to a systemic impairment of T cells necessary for the development of an effective anti-cancer response. Taking into account the recent development of inhibitors of amino acid metabolic pathways, we also assume that inhibition of the enzymatic activity of the above-mentioned enzyme system may be a target for novel anti-cancer strategies, especially in combination with the already existing targeted therapies at the molecular level, but also with classical chemotherapy.
The project aims to investigate the mechanism of the development of endometriosis - a common disease of women causing pain, infertility and sometimes leading to the development of ovarian cancer. In the course of the disease, endometrioid epithelial tissue develops ectopic, most often in the peritoneal cavity. The most widely recognized theory of disease development postulates that foci of ectopic endometrioid tissue originate in the eutopic endometrium, but many aspects of the disease remain unexplained. One of the unconfirmed hypotheses is the accumulation of somatic mutations in eutopic endometrial epithelial cells. It has been proven that in the course of endometrial cancer of the ovary there is microdeletion in the cells of endometriotic foci, which then give rise to the tumor, while in endometriotic lesions not related to cancer, no such microdeletions were found. In the proposed project, it is planned to study the sequences of exomalous tissues from women suffering from endometriosis and from healthy women. Based on the analysis of the obtained mutation pattern, we plan to determine whether there are somatic mutations in the ectopic endometrium that may be the cause of ectopic changes.
It is known that the proper functioning of the human body depends largely on the bacteria that are found in the intestines. Recent studies indicate that disturbances in the composition of the intestinal flora are associated with arterial hypertension. The bacteria in the gut produce a number of chemicals that enter the bloodstream, including hydrogen sulfide (H2S) and methane. The aim of the proposed project is to investigate the effect of increased concentrations of H2S and methane in the intestines on the regulation of blood pressure and the development of hypertension. The research will also produce new compounds that can release H2S. The research will be carried out on rats. 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 will help elucidate the role of metabolites produced by gut bacteria in the regulation of blood pressure and in the pathogenesis of hypertension. The study will help assess whether drugs that alter the concentration of H2S and methane in the intestines can be used in the treatment of cardiovascular diseases, in particular arterial hypertension.
The aim of the study is to find out the influence of intestinal bacteria and the methylamines they produce on the development of arterial hypertension. Recent studies indicate that arterial hypertension may be associated with disturbances in the composition of the intestinal flora. The effect of gut bacteria on the functioning of the circulatory system can take place through a number of chemicals produced by the gut microflora that enter the bloodstream. More and more studies suggest that methylamines produced by intestinal bacteria may play an important role in the pathogenesis of cardiovascular disease. It has been shown that an increase in the concentration of trimethylamine oxide in the blood is associated with a higher risk of myocardial infarction, stroke and death. In the proposed project, tests will be carried out on rats fed with water or water containing the tested methylamines. The effect of methylamines on blood pressure and heart function will be investigated. Hypertension and its complications are one of the most important causes of death and disability. The proposed project aims to investigate new compounds that may be responsible for the development of hypertension. The results of the study may contribute to the creation of a new group of drugs for arterial hypertension, the mechanism of action of which will be based on the modification of the production of methylamines in the human intestines.
Acute B-cell lymphoblastic leukemia (B-OBL) is a biologically diverse group of diseases that underlie specific genetic changes that result in abnormal differentiation and proliferation of the hematopoietic cell. In recent years, there has been great progress in the treatment of B-OBL, especially in children. Unfortunately, the results of treatment in adults are still unsatisfactory. One of the reasons for this may be the presence of two genetic subtypes of leukemia with a very high risk of disease progression in more than half of adults with B-OBL - BCR-ABL1 positive and BCR-ABL1-like. Both leukemia subtypes, despite the different genetic changes occurring in them, show a similar gene expression profile, which consequently leads to the activation of the JAK-STAT pathway. Recent reports indicate that both BCR-ABL1-positive B-OBL and BCR-ABL1-like may share a similar mechanism responsible for disease progression, which is related to an unfavorable prognosis. Both BCR-ABL1-positive and BCR-ABL1-like leukemia have been shown to be characterized by a high percentage of specific genetic mutations (changes in copy number of DNA fragments; CNAs), mainly deletions. Their presence is probably due to abnormal activity of some cellular enzymes. The complex of recombinases - RAGs and proteins from the APOBEC family, including cytidine deaminase AID, are enzymes involved in a very important physiological process, namely the production of functional antibodies and the formation of their diversity in contact with the pathogen. The activity of both enzymes is characteristic of the lymphoid tissues from which B-OBL is derived. However, it is a double-edged sword, which, on the one hand, allows the body to produce antibodies that protect us against the threat of microbes, and on the other hand, it can cause genetic changes leading to the formation and progression of cancer. Therefore, the project focuses on endogenous genomic mutators such as RAGs, AID, APOBEC3A, APOBEC3B. For this purpose, various methods of molecular biology will be used, including DNA sequencing and gene expression analysis. The project aims to elucidate the molecular basis leading to a poor prognosis in adults with B-OBL, namely the phenomena of genetic instability mediated by RAGs and proteins from the APOBEC family. During genetic instability, mutations accumulate, favoring the selection of leukemic clones and causing disease progression. Despite the great progress in this field of science, there is still not enough knowledge about it. The above research may also translate into other areas of oncology, constituting a starting point and inspiration to repeat similar research in other disease entities.