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Circulating tumor cells in personalized cancer therapy
Jakabová, Anna ; Kološtová, Katarína (advisor) ; Svoboda, Bohuslav (referee) ; Tencerová, Michaela (referee)
Circulating tumor cells (CTC) have showed great potential to become both prognostic and predictive biomarker in various types of oncological diseases. CTC can help detect patients in higher risk of shorter overall survival, progression-free survival or relapse. They can be also helpful in therapy selection as in current clinical practice treatment is chosen based on primary tumor characteristics. Regular CTC counts and features monitoring can be real-time indication of therapy response and can be used to guide-targeted treatment. This information can be implemented to personalized medicine and each cancer patient can be treated based on individual profile. However, patients' sample with CTC is easily accessible, their detection has remained challenge due to low CTC number in the circulation and heterogeneous nature. CTC can circulate in blood in the form of single cells or in clusters that usually represent minority in comparison with single CTC but their metastatic potential is significantly greater than of single CTC. Apart from CTC count, the molecular character showed dynamic development and heterogeneous nature not only between patients but also within the individual patient's tumor tissue itself. Character of primary tumor, CTC and metastasis are not always consistent and has been changing...
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Long-term monitoring of ctDNA levels in patients with metastatic colorectal cancer for early detection of progression or recurrence of the disease
Kopalová, Dominika ; Benešová, Lucie (advisor) ; Kološtová, Katarína (referee)
Circulating tumor DNA (ctDNA) in peripheral blood of patients with metastatic colorectal cancer appears to be a promising molecular marker that provides various applications. ctDNA levels vary depending on the presence, alternatively on the volume of tumor mass within patient's body, which can be used primarily for early detection of disease progression or recurrence and moreover for evaluating radicality of surgical treatment, all within long-term postoperative follow-up of the patient. Due to minimal invasivity of ctDNA analysis from peripheral blood (so-called liquid biopsy), it is possible to perform it repeatedly at relatively short time intervals. On account of very low fraction of ctDNA in total cell-free DNA (cfDNA) ranging between units and hundreds of percent, the key factor is optimal methodology covering all steps from the isolation process to a sufficiently sensitive detection technology. In this thesis I focus on an optimization of isolation process and analysis of ctDNA obtained from tumor tissue and plasma of selected patients with metastatic colorectal cancer in connection with surgical radicality and correlation with a clinical status of the patients.
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TaqMan-based nucleic acid quantification - abilities and limits with regards to type of collection, age and quality of human specimen
Herzogová, Eva ; Daňková, Pavlína (advisor) ; Kološtová, Katarína (referee)
Real-time PCR method is a type of PCR which allows continual monitoring of DNA amplification during every cycle of its process. It is mostly used for gene expression analysis. Based on the results of previous experiments, we decided to test out the effect of anticoagulants EDTA, heparin, sodium citrate and CPDA on the expression of selected genes of the immunological spectrum and further, to test how the time period between drawing the blood and processing of blood sample influences mRNA levels of selected genes that are determined by changes in gene expression and/or mRNA degradation. To quantify mRNA of the studied genes, we isolated total RNA from the peripheral blood leucocytes and transcribed it into cDNA by using the reverse transcription PCR. This cDNA served as a template for the real-time PCR. To examine the changes of the expression caused by the effect of each particular anticoagulants, peripheral blood derived from 10 volunteers was used (each donor's blood was taken into 3 vacuum tubes with EDTA, heparin and sodium citrate anticoagulant agents). Next to that, we obtained 10 buffy coat samples in transfusion blood bags with CPDA anticoagulant agent. Compared to blood cells influenced by one of the three anticoagulant agents present in vacuum tubes, cells from transfusion bags affected...
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Immunogenetic studies on autoimmune diabetes mellitus
Kološtová, Katarína ; Černá, Marie (advisor) ; Mateička, František (referee) ; Štechová, Kateřina (referee)
Immunogenetic studies on autoimmune diabetes. Aims of the study: The study has to characterize the genetic background of patients with different types of diabetes mellitus (T1D in children, T1D in adults, LADA, T2D, MODY). The relationship of the diabetes associated HLA-DRB1*04 and NFKB1 genes to the disease course was proved further in the functional studies of the mRNA gene expression. Patients were divided into the tested subgroups in relation to the HLA class II, NFKB1, and NFKBIA genotypes and disease type (T1DM in children, T1DM in adults, and LADA). Results and Conclusion: According to our findings we can conclude that the progression of the diabetes in T1D adults, T1D children and LADA is strongly influenced by different immunogenetic background modifying the ethiopathogenesis of diabetes in the above described groups. Our results offer new possibilities for the population risk testing and may be that far used in the future for better diagnostics of the diabetic's adults.
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