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Effect of TGFβ on the production of extracellular matrix proteins by mesenchymal cells in the glioblastoma microenvironment
Vepřková, Jana ; Bušek, Petr (advisor) ; Přechová, Magdalena (referee)
Glioblastoma (GBM) is the most aggressive and common malignant primary brain tumor, characterized by extracellular matrix (ECM). These changes may be caused by transforming growth factor beta (TGFβ), which is overexpressed in GBM. TGFβ induces the expression of fibroblast activating protein (FAP), which is found in GBM on mesenchymal cells surrounding blood vessels, where ECM proteins collagen type I and fibronectin are typically found. This study aimed to determine the effect of TGFβ on the expression of collagen type I and fibronectin by FAP+ mesenchymal cells in the GBM and to determine the effect of ECM produced by these cells on glioma cell migration. Bioinformatic and tissue analyses demonstrate a correlation between TGFβ, FAP, collagen type I, and fibronectin expression in GBM. Using immunohistochemistry, TGFβ expression was detected diffusely in GBM including areas of stroma that contained FAP+ mesenchymal cells and collagen type I. In in vitro experiments, FAP+ mesenchymal cells showed increased production of collagen type I and fibronectin after TGFβ stimulation. In contrast, glioma cells showed very low production of these proteins even after TGFβ stimulation. ECM produced by FAP+ mesenchymal cells significantly increased the migration of the glioma cell line U251. However, no further...
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Role fibroblastového aktivačního proteinu v růstu a invazivitě gliomových buněk
Fejfarová, Edita ; Bušek, Petr (advisor) ; Szabo, Pavol (referee)
High grade astrocytomas are very progressive brain tumors. Glioblastoma multiforme is the most frequent and the most malignant type with very infiltrative phenotype of the tumor cells. Fibroblast activation protein FAP is a predominantly membrane bound prolyl peptidase bearing exo- and endopeptidase hydrolytic activities. FAP is known to play a role in wound healing, cell migration and invasion and its expression is linked to the pathogenesis of several malignancies. mRNA expression of FAP is upregulated in 48% of glioblastomas according to The Cancer Genome Atlas microarray data. The involvement of FAP in the pathogenesis of astrocytic tumors is largely unknown. The aims of this work are to analyse the expression of FAP in primary cell cultures derived from high grade gliomas and to analyse the influence of FAP on the growth, migration and invasion of glioma cells. Our ELISA and western blot results showed heterogenous expression of FAP in the studied glioma primary cell cultures and cell lines. Both enzymatic activities characteristic of FAP were detected in the primary glioma cell culture P11 with high expression of FAP. In these cells, FAP was present not only in the typical plasma membrane localization, but also in the cytoplasm as demonstrated by immunofluorescence staining. The P11 cells...
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The role of dipeptidyl peptidase-IV and homologous proteases in migration and invasion.
Fejfarová, Edita ; Bušek, Petr (advisor) ; Rösel, Daniel (referee)
Migration and invasion are processes which naturally occur in organism during embryogenesis, immune reactions or wound healing. These processes are very important factors in some serious diseases like rheumatoid arthritis and carcinogenesis. There is no doubt about contribution of proteases in these processes-many of them degrade extracellular matrix and thereby facilitate the movement of cells. While dipeptidylpeptidase-IV cleaves solely two amino acids from N-terminus so it is not considerably involved in ECM degradation. DPPIV and its homologues recognize peptides with proline on penultimate position, which causes resistance to ordinary types of proteases. Substrates of DPPIV and its activity homologues include chemokines implicated in signalling of migration - their cleavage and thus inactivation present DPPIV and DPPIV-like molecules as modulators of cell migration signalling in choriocacinoma, neuroblastoma, on Sézary cells or epithelial cells migrating in response to injury. Another activity of some DPPIV-like proteases is binding to the extracellular matrix proteins, when are helping in the attachment of cells and thus affect the migratory ability of the cells like ovarian cancer cells, prostate cancer cells, melanoma cells or kidney cells. Effects on migration and invasion have also...
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Development of nanochemical tools targeting receptors in the tumor microenvironment
Blažková, Kristýna ; Konvalinka, Jan (advisor) ; Abramson, Jakub (referee) ; Bušek, Petr (referee)
Development of nanochemical tools targeting receptors in the tumor microenvironment Targeting the receptors in the tumor microenvironment is crucial for the future development of targeted therapies, precision medicine and immunotherapy of cancer. The options available now are, however, limited by the availability of specific ligands. The advances in the field strongly rely on the use of antibodies and genetic modifications of immune cells. Availability of small molecules targeting the receptors of interest would allow further development of alternative strategies as well as deepen our understanding of the underlying mechanisms of cancer development, progression and clearance. In the search for new small-molecule ligands and their use for receptor targeting, the prostate-specific membrane antigen (PSMA) and the immune receptors CD3 and CD64 were selected as model targets. The selected method - the phage display of bicyclic peptides - utilizes chemical modification of the displayed three-cysteine peptides to achieve their cyclization and formation of bicycles. The panning of a peptide library displayed on the phages and probed with PSMA revealed a reproducibly-selected amino acid sequence. Interestingly, the phage clone carrying this sequence was a specific binder of PSMA, but the synthesized peptide alone...
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Implementation of logistics modules of information system in Niedax s.r.o.
BUŠEK, Petr
The topic of this diploma thesis is focused on implementation of logistic modules of the enterprise information system in the Niedax company. The theoretical part provides information from the field of information systems, supplemented by information about ERP (Enterprise Resource Planning), IS/IT expenses and methods of design and implementation of information systems. The practical part briefly describes the selected information system, the company where the information system was implemented, the implementation partner and the BMD information system. Furthermore, and analysis of the current state is carried out and the course of the whole implementation project is described, followed by a practical demonstration of the BMD system. Finally, an evaluation of the project is performed. Evaluation is based in a comparison of the TCO (Total Cost of Ownership) values determined in the practical part of the thesis and an evaluation of the questionnaire survey.
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The Role of the Tumour Microenvironment on Melanoma Cell Invasiveness
Jobe, Njainday ; Rösel, Daniel (advisor) ; Kořínek, Vladimír (referee) ; Bušek, Petr (referee)
Cancer cell invasion and metastasis are hallmarks of cancer. It is becoming apparent that the interaction between cancer cells and the surrounding microenvironment are involved in their ability to invade and metastasise. In general, cancer cells can either migrate individually, in an amoeboid or mesenchymal manner, or collectively. The first aim of this thesis was to analyse the role of NG2 in amoeboid to mesenchymal transition (AMT) and Rho/ROCK signalling. We found that NG2 promotes an amoeboid morphology, and increased invasiveness, in a Rho-dependent manner. Secondly, we analysed the role of the major tumour microenvironment (TME) component, cancer-associated fibroblasts (CAFs), on melanoma cell invasiveness. We found the CAF interaction with melanoma cells leads to increased levels of interleukin-6 (IL-6) and IL-8, and this leads to increased invasiveness. Simultaneous blocking of IL-6 and IL-8, using neutralising antibodies, inhibits CAF-dependent invasion. Further analysis of another major component in the melanoma TME, keratinocytes, has highlighted the importance of the tumour cell niche in invasion. Our results indicate that cancer cells have the ability to change morphology, and that the TME plays an important role in melanoma cell invasiveness. Metastatic melanoma treatment has proven...
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Role fibroblastového aktivačního proteinu v růstu a invazivitě gliomových buněk
Fejfarová, Edita ; Bušek, Petr (advisor) ; Szabo, Pavol (referee)
High grade astrocytomas are very progressive brain tumors. Glioblastoma multiforme is the most frequent and the most malignant type with very infiltrative phenotype of the tumor cells. Fibroblast activation protein FAP is a predominantly membrane bound prolyl peptidase bearing exo- and endopeptidase hydrolytic activities. FAP is known to play a role in wound healing, cell migration and invasion and its expression is linked to the pathogenesis of several malignancies. mRNA expression of FAP is upregulated in 48% of glioblastomas according to The Cancer Genome Atlas microarray data. The involvement of FAP in the pathogenesis of astrocytic tumors is largely unknown. The aims of this work are to analyse the expression of FAP in primary cell cultures derived from high grade gliomas and to analyse the influence of FAP on the growth, migration and invasion of glioma cells. Our ELISA and western blot results showed heterogenous expression of FAP in the studied glioma primary cell cultures and cell lines. Both enzymatic activities characteristic of FAP were detected in the primary glioma cell culture P11 with high expression of FAP. In these cells, FAP was present not only in the typical plasma membrane localization, but also in the cytoplasm as demonstrated by immunofluorescence staining. The P11 cells...
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The role of dipeptidyl peptidase-IV and homologous proteases in migration and invasion.
Fejfarová, Edita ; Rösel, Daniel (referee) ; Bušek, Petr (advisor)
Migration and invasion are processes which naturally occur in organism during embryogenesis, immune reactions or wound healing. These processes are very important factors in some serious diseases like rheumatoid arthritis and carcinogenesis. There is no doubt about contribution of proteases in these processes-many of them degrade extracellular matrix and thereby facilitate the movement of cells. While dipeptidylpeptidase-IV cleaves solely two amino acids from N-terminus so it is not considerably involved in ECM degradation. DPPIV and its homologues recognize peptides with proline on penultimate position, which causes resistance to ordinary types of proteases. Substrates of DPPIV and its activity homologues include chemokines implicated in signalling of migration - their cleavage and thus inactivation present DPPIV and DPPIV-like molecules as modulators of cell migration signalling in choriocacinoma, neuroblastoma, on Sézary cells or epithelial cells migrating in response to injury. Another activity of some DPPIV-like proteases is binding to the extracellular matrix proteins, when are helping in the attachment of cells and thus affect the migratory ability of the cells like ovarian cancer cells, prostate cancer cells, melanoma cells or kidney cells. Effects on migration and invasion have also...
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The Role of Dipeptidyl Peptidase-IV in Glioma Cell Growth
Bušek, Petr ; Šedo, Aleksi (advisor) ; Trka, Jan (referee) ; Konvalinka, Jan (referee)
Dipeptidyl peptidase-IV (DPP-IV, CD26, EC 3.4.14.5) is a widely expressed serine protease that by limited proteolysis regulates a number of biologically active peptides including a number of mitogenic peptides involved in cancer development. Deranged DPP-IV expression and/or enzymatic activity has been reported in a number of tumors, and could lead to altered signaling and biological function of its substrates. Changes in DPP-IV expression were observed in glioma cell lines in association with differentiation, and recently also in malignant gliomas in vivo. In addition, DPP-IV substrates substance P (SP) and stromal cell derived factor-1 (SDF-1) that trigger growth promoting intracellular signaling cascades in glioma cells have strongly been implicated in the pathogenesis of gliomas. The aim of this study was (i) to investigate the effects of DPP-IV on the signaling of its biologically active substrates that promote the malignant phenotype of glioma cells, and (ii) to assess the effects of DPP-IV on the growth, migration and adhesion of human glioma cells. Using transfected glioma cell lines (U373CD26, T98GCD26, U87CD26) with mifepristone-inducible DPP-IV expression, we demonstrate that DPP-IV overexpressing T98GCD26 cells can cleave SP and thus abrogate its ability to trigger calcium signaling in U373...
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Implementation of enterprise information system in the selected company
BUŠEK, Petr
This thesis deals with the implementation of enterprise information system in the selected company. The theoretical part provides information from the field of information systems, which are complemented by the analysis of internal and external environment, the strengths and weaknesses, and the method of selection of the information system. In the practical part briefly describes the selected information system and the company, where the information system was implemented. Furthermore, the current information system is analyzed by the HOS 8 method, from which the targets for the implementation of a new information system was set. At the end of the practical part I will compare the system before implementation and the system after implementation with the HOS 8 analysis, which is the main goal of this thesis.
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