National Science Centre

Increased anisocytosis of erythrocytes expressed by the parameter: red blood celldistribution width (RDW) is associated with an increased risk of myocardial infarction and stroke. It is also an unfavorable prognostic factor after stroke, myocardial infarction, in patients with diabetes or chronic ischemia of the lower limbs. The aim of the project is to determine whether erythrocyte heterogeneity is only a new indicator of atherosclerosis and a prognostic factor for its complications, or whether it is a new factor in the remodeling of the vessel wall. The study also aims to identify the mechanisms by which erythrocyte anisocytosis affects the development of atherosclerosis. The implementation of the project will allow the identification of new indicators that can be used in the future to assess the risk of atherosclerosis. It will also enable the development of potential new therapeutic methods. Most scientists see the phenomenon of anisocytosis associated with generalized inflammation as a prognostic factor for these pathological conditions. Our hypothesis is that the phenomenon of anisocytosis is a predictor of the aforementioned cardiovascular dysfunctions. Its validity will be checked as part of the project.

Hepatitis C virus (HCV) infection is widespread in the world - the number of infected people is estimated at 170 million. About 50% of infected people complain of chronic fatigue, depression, difficulties with concentration, and impaired neurocognitive function. These symptoms are independent of both the degree of liver damage and the level of viral replication. HCV was long considered an exclusively hepatotropic virus, but there is now strong evidence that it is also lymphotropic and that it grows in peripheral blood mononuclear cells (PBMCs), among others. Posthepatic HCV replication can have serious clinical implications: infected leukocytes that cross the blood-brain barrier transmit the infection to the central nervous system (CNS). Hypothesis. Our general hypothesis is that HCV infects PBMCs, particularly monocytes / macrophages, and that infected cells cross the blood-brain barrier. Being in the CNS, they release pro-inflammatory cytokines that cause functional changes manifested by depression and neurocognitive disorders. The hypothesis predicts that the immune activation state of PBMCs correlates with the severity of these disorders. Moreover, the mere presence of HCV replication in PBMCs may indicate possible brain infection and may correlate with neurocognitive disorders and depression. The current project aims to investigate the relationship between HCV replication in PBMC and the activation status of these cells, and neurocognitive disorders in patients with chronic hepatitis C.

Epilepsy is a condition whose nature is still poorly understood. Despite the conducted research, it is still not possible to identify the reasons for the increased sensitivity of some people to convulsive factors, as well as the reasons why standard antiepileptic therapy is not effective in a certain percentage of patients. The aim of our research is to try to analyze the basis of individual differences in the rate of development of seizures, as well as differences in response to antiepileptic drugs. The analysis will include the assessment of biochemical parameters (concentrations of monoamines, inhibitory and excitatory amino acids) and immunocytochemical parameters (expression of selected receptor subunits, inflammatory mediators and markers of neuroplastic remodeling). An important element will also be the analysis of the expression of the REST protein involved in the regulation of GABAergic activity and mTOR kinase activity, the role of which in the pathogenesis of epilepsy associated with tuberous sclerosis is indisputable; however, its possible participation in the pathogenesis of acquired epilepsy is also postulated at present. We also plan to analyze the effectiveness of antiepileptic drugs in selected populations with different susceptibility to the development of seizures.

The knowledge about the phenomena accompanying the earliest stages of hepatitis C virus (HCV) infection is still unsatisfactory. This is due to the lack of a good experimental model and the specificity of the clinical course of the infection, which at this stage is usually asymptomatic. In particular, the knowledge about the factors that determine the elimination of a virus or its survival and disease development is still incomplete. In the presented Project, we plan to study 180 blood donors with newly acquired HCV infection. The aim of the presented project is to investigate the factors influencing the early stage of infection that determine the elimination or survival of HCV infection. Factors from both the host side: activation of the immune system response and genetic conditions (IL28B genotype), as well as from the virus side: population complexity and the occurrence of characteristic HCV HVR1 and 5'UTR sequence motifs using deep sequencing techniques will be analyzed.

Diabetic foot syndrome (DFU) - is a serious complication of long-lasting diabetes, occurring in more than 15% of patients with diabetes mellitus (ncuropathy and peripheral arterial diseases are the main factors of phalcyclic / SC). Depending on the predominant etiological factor, DFU is divided into types: neuropathic DFU, vascular DFU and mixed type. The research hypothesis is the existence of variability within the Osteoprotegerin gene affecting the frequency of particular types of diabetic foot syndrome. The aim of the study is to evaluate the role of OPG polymorphisms in patients with type 2 diabetes, diabetic foot syndrome of neuropathic, vascular and mixed etiology and Charcot neuroarthropathy.

1. Identification of new, rare and functional genetic variants within genes related to the process of activation and aggregation of platelets in the Polish population with a history of ischemic stroke. 2. Compared carriers of rare functional variants within individual genes (potentially harmful) in the study group and the control group (without a history of ischemic stroke) in order to determine their relationship with the risk of stroke. 3. Assessment of the relationship between the presence of rare functional variants and short-term prognosis based on the assessment of neurological disorders in patients after ischemic stroke. 4. Functional studies of genes based on the use of in vitro cell culture as an experimental model to confirm changes in protein activity at the cellular level.

The research hypothesis of the project assumes that the production of two key mediators in the trigeminal system - the calcitonin gene derivative peptide (CGRP) and the cerebral neurotrophic factor (BDNF) depends on the activity of neurons and the presence of inflammatory factors. The aim of the project is to investigate the role of the pro-inflammatory cytokine - tumor necrosis factor (TNF) in modulating the function of sensory neurons in the trigeminal ganglion.

The γc receptor (CD132) is a subunit of interleukin (IL) 2, 4, 7, 9, 15 and 21 receptors. Too high activation of γc leads to an excessive immune response and may lead to the development of autoimmune diseases. The binding of a ligand to the γc receptor leads to the phosphorylation of JAK1 and JAK3 Janus kinases, followed by signal transduction to the nucleus and activation of gene expression. In recent years, research has been conducted into the possible use of JAK1 and JAK3 kinase inhibitors in the treatment of autoimmune diseases. The aim of the research planned in this project is to investigate whether cefazolin, a cephalosporin antibiotic, inhibits γc receptor activation.

The aim of this study is to identify prognostic protein markers for vulvar squamous cell carcinoma (VC). Computerized analysis of changes at the level of regulatory pathways to model carcinogenesis could provide new strong hypotheses about the biological causes of VC progression. We intend to develop a method for quantifying the expression of a selected panel of proteins using the MRM technique (multiple reaction monitoring). The developed test will enable the quantification of the expression of the studied proteins. Correlation of the level of peptides in tumors with clinical data obtained during the observation period will allow the assessment of the predictive value of the test. The research methodology proposed in this application may lead to the detection of many proteins with different expression in tumors obtained from patients with progressive VC ("progVC") during the follow-up period compared to the expression in tumors of patients with long disease-free survival ("d-fVC" ”) During 8-12 years of observation. These proteins can then be grouped according to common biological characteristics, revealing the changes that cause the tumor's aggressive phenotype.

The project is carried out as part of the scientific consortium. The leader of the project is the Institute of Human Genetics of the Polish Academy of Sciences, while the partners are the M. Nencki Institute of Experimental Biology of the Polish Academy of Sciences and the Medical University of Warsaw, 1st Faculty of Medicine. In the proposed project, we assume that in tumors, apart from the already well-documented gene damage, there are also unknown mutations within the regulome (non-coding sequences responsible for the regulation of gene expression), which significantly contribute to the pathogenesis of these diseases. The aim of this project is to identify potential mutations within the regulome of laryngeal squamous cell carcinoma (LSCC). In this project, we plan targeted NGS sequencing of the regulome, which includes a defined region of 46 min base pairs, covering the proximal enhancers and promoters, identified experimentally with DNase l-seq and CAGE methods in the ENCODE and FANTOM projects; both constitutive (active in many cell types) and epithelial specific. Sequencing will be performed on the HiSeq 1500 genomic platform (Illumina) using the TruSeq SBS Kit (Illumina).All libraries will be sequenced in paired end mode (2 x 100 bp) with an average coverage of 90x (minimum 20x) for reference samples and 190x (minimum 40x) for cancer cell lines and tumors. The tested material, 94 samples in total, will consist of 7 pairs: LSCC cell line - belonging culture of non-neoplastic fibroblasts and 40 pairs: pharyngeal tumor of the larynx - peripheral blood (from the same patient). At the same time, genome-wide analysis of gene expression using microarrays will be carried out in a group of 7 tested cell lines (a total of three repetitions) and in a group of 15 non-cancer reference samples (epithelial cells). The obtained results will be subjected to bioinformatics analysis, the purpose of which will be to identify somatic changes by comparing the cell line - fibroblasts, cancer - blood, and then integrating the identified changes with expression data. Such an approach will allow the emergence of mutations in regulatory regions in the vicinity of genes with significantly altered expression. For this purpose, a constantly updated database of regulatory areas and motives (Nencki Genomics Database - NGD) will be used. Thus, the selection of the most important changes in regulatory sequences will be carried out on the basis of: (a) the impact of the changes found on the expression of neighboring genes (comparison of expression in groups of cell lines - control non-cancer tissues, microarray data), (b) frequency of changes in a given regulatory sequence ( in a group of 40 tumors). In the last stage of the work, the most important, identified changes will be validated by other techniques (pyrosequencing, Sanger sequencing, qRT-PCR) in the group of tested cell lines and primary tumors, as well as by Western blot at the protein level. In addition, targeted testing of the identified loci with the above techniques in an additional group of 50 primary laryngeal tumors will be performed to estimate the frequency of lesions. As of today, there are no reports in the literature on the role of regulome changes in the pathogenesis of LSCC. Hence, the planned research is the first research approach on a global scale that integrates data on the presence of regulatory sequence variants with global gene expression in these tumors. We believe that this research approach may contribute to the description of significant changes in the regulom, which will contribute to a better understanding of the genetic basis of LSCC pathogenesis.