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Biomarker detection using electrochemical method with microfluidic chip
Klepáčová, Ivana ; Svoboda, Ondřej (referee) ; Neužil, Pavel (advisor)
The thesis is focused on the development of the electrochemical system with microfluidic platform for the detection of multiple biomarkers. It analyses the use of biomarkers for the early diagnosis of cancer. The theoretical part contains basic information about voltammetric methods and microfluidic systems. The practical part provides solutions to the microfluidic chips, including the description of the used materials, designs, methodologies of preparation and conclusions from the testing of the manufactured microfluidic systems. The thesis describes the lock-in electrochemical system which measures the response of 4 electrochemical cells simultaneously. For the electrochemical system measurements, an electrochemical chip consisting of 64 electrochemical cells was used. The results of the analysis include the processing of the system tests and detected voltammetric curves of the Fe2+/Fe3+ solution and cysteine.
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Therapeutic use of alternative protein binders targeting tumor biomarkers in clinical testing of oncology patients
Tauš, Petr ; Drbal, Karel (advisor) ; Lepšík, Martin (referee)
Almost until the end of the last century, antibodies (aka immunoglobulins) were considered the only class of specific binding proteins. The discovery of hybridoma technology in 1975 had enabled the production of monoclonal antibodies and after twenty years some of them have entered clinical practice. Meanwhile, the first non-immunoglobulin protein scaffold, in which new specific binding sites could be introduced was discovered. To date, many different alternative scaffolds have been described, but only a few of them are being further developed for diagnostics, therapeutics or tools in basic research. Since these structures are overcoming the drawbacks of immunoglobulin structure, which are big size, expensive production and difficult rational design, they have potential to replace and exceed them. In this bachelor's thesis all the alternative scaffolds in development are summarized. Moreover, their advancements in clinical trials are described and compared with approved therapeutics based on immunoglobulin structure.
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Antigenome defines a selection of mutated tumor peptides driving tumor-specific T-cell response
Hadlová, Petra ; Drbal, Karel (advisor) ; Dibus, Michal (referee)
T cells, as an essential part of the adaptive immune system, play crucial role in eradication of tumor growth. T cells target, interact with and eventually annihilate the tumor cells in antigen- specific (Ag) manner. T cells interact with tumor cells via short epitopes bound to the major histocompatibility complex (MHC) molecules on the tumor cell surface. Tumor specific neoepitopes arise from random somatic mutations and constitute a part of the tumor antigenome. Antigenome comprises of two classes of antigens, tumor specific antigens (TSA) and tumor associated antigens (TAA). TSA are neoantigens carrying neoepitopes unique to each tumor. TAA are self-antigens presented by both tumor cells and non-transformed cells. Each tumor cell is able to develop numerous ways to evade the immune system consisting of T cells, NK cells, macrophages and other mechanisms employed. Despite that immunotherapy has shown a great potential in personalized medicine. The stratification of responsive patients is essential for effective and durable management of therapy in clinical practice. Methods are employed, which study existing reactive T cell clones, somatic mutations present in each patient, role of somatic mutations in tumor development and present neoepitopes. All these patient- specific features facilitate...
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Somatic driver mutations during early differentiation of bladder carcinoma cell of origin
Brázdilová, Ludmila ; Drbal, Karel (advisor) ; Láníková, Lucie (referee)
A normal healthy cell traves through different routes to become a tumor cell, which according to the cell-of-origin theory initiates the whole tumor. Deregulation of cell processes by somatic mutations directs the cell into transformation. To this day, many mutations that cause a tumor phenotype, termed driver mutations, have been identified by genomic and targeted analyses. Not only for optimal therapy management but also for the prediction of disease progression the detection of driver mutations accumulating in the cell of origin of a specific tumor is very important. This thesis is focused on driver mutations of bladder carcinoma cell of origin, which is a tumor with a high mutation load. Bladder carcinomas compose a very heterogeneous group of tumors, having phenotype parallels in many other carcinomas, such as breast cancer. The driver mutations could be used as diagnostic and prognostic markers, but are not yet used in clinical practice. This thesis intends to summarize known findings about bladder carcinoma tumor initiation, based on understanding its cell of origin. Further characterisation of important driver mutations in bladder carcinoma and a comparison to other carcinomas is shown here, with respect to their molecular classification.
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Antigenome defines a selection of mutated tumor peptides driving tumor-specific T-cell response
Hadlová, Petra ; Drbal, Karel (advisor) ; Dibus, Michal (referee)
T cells, as an essential part of the adaptive immune system, play crucial role in eradication of tumor growth. T cells target, interact with and eventually annihilate the tumor cells in antigen- specific (Ag) manner. T cells interact with tumor cells via short epitopes bound to the major histocompatibility complex (MHC) molecules on the tumor cell surface. Tumor specific neoepitopes arise from random somatic mutations and constitute a part of the tumor antigenome. Antigenome comprises of two classes of antigens, tumor specific antigens (TSA) and tumor associated antigens (TAA). TSA are neoantigens carrying neoepitopes unique to each tumor. TAA are self-antigens presented by both tumor cells and non-transformed cells. Each tumor cell is able to develop numerous ways to evade the immune system consisting of T cells, NK cells, macrophages and other mechanisms employed. Despite that immunotherapy has shown a great potential in personalized medicine. The stratification of responsive patients is essential for effective and durable management of therapy in clinical practice. Methods are employed, which study existing reactive T cell clones, somatic mutations present in each patient, role of somatic mutations in tumor development and present neoepitopes. All these patient- specific features facilitate...
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Cell surface CD47 expression in cancer stem cell-targeted tumor therapy
Kuzmík, Ján ; Drbal, Karel (advisor) ; Černý, Jan (referee)
CD47 is a transmembrane glycoprotein with a high expression in both, healthy and cancer (stem) cells. Level of the CD47 expression is negatively correlated with survival of cancer patients. Binding of CD47 to SIRPα, localized on a phagocyte, triggers intracellular signaling cascade. The final effect of this cascade is dephosphorylation of nonmuscle myosin-IIA, which disrupts its function and accumulation to phagocytic synapse. The blockage of CD47-SIRPα signaling pathway in a presence of the pro-phagocytic signal induces phagocytosis of cancer cells. Afterwards, phagocytes can serve as the antigen presenting cells and prime T cell response. Role of CD47-SIRPα signaling pathway in immunity has established this pathway as a target of cancer therapy testing. Preclinical research has identified a positive therapeutic effect of blocking this signaling pathway. Nowadays, the first phase of clinical trials is being conducted. The most prevalent approach of blocking CD47-SIRPα signaling pathway in therapy is the use of anti-CD47 blocking monoclonal antibodies, which cause mild anemia. However, alternative approaches of blocking this pathway are also being developed. In this bachelor thesis, I have summarized the research related to the blockage of CD47-SIRPα signaling pathway as a cancer therapy.
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Therapeutic use of alternative protein binders targeting tumor biomarkers in clinical testing of oncology patients
Tauš, Petr ; Drbal, Karel (advisor) ; Lepšík, Martin (referee)
Almost until the end of the last century, antibodies (aka immunoglobulins) were considered the only class of specific binding proteins. The discovery of hybridoma technology in 1975 had enabled the production of monoclonal antibodies and after twenty years some of them have entered clinical practice. Meanwhile, the first non-immunoglobulin protein scaffold, in which new specific binding sites could be introduced was discovered. To date, many different alternative scaffolds have been described, but only a few of them are being further developed for diagnostics, therapeutics or tools in basic research. Since these structures are overcoming the drawbacks of immunoglobulin structure, which are big size, expensive production and difficult rational design, they have potential to replace and exceed them. In this bachelor's thesis all the alternative scaffolds in development are summarized. Moreover, their advancements in clinical trials are described and compared with approved therapeutics based on immunoglobulin structure.
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Biomarker detection using electrochemical method with microfluidic chip
Klepáčová, Ivana ; Svoboda, Ondřej (referee) ; Neužil, Pavel (advisor)
The thesis is focused on the development of the electrochemical system with microfluidic platform for the detection of multiple biomarkers. It analyses the use of biomarkers for the early diagnosis of cancer. The theoretical part contains basic information about voltammetric methods and microfluidic systems. The practical part provides solutions to the microfluidic chips, including the description of the used materials, designs, methodologies of preparation and conclusions from the testing of the manufactured microfluidic systems. The thesis describes the lock-in electrochemical system which measures the response of 4 electrochemical cells simultaneously. For the electrochemical system measurements, an electrochemical chip consisting of 64 electrochemical cells was used. The results of the analysis include the processing of the system tests and detected voltammetric curves of the Fe2+/Fe3+ solution and cysteine.
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