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Immunoscore in 3D tissue
Novák, Jaromír ; Drbal, Karel (advisor) ; Procházka, Jan (referee)
Solid tumors are complex structures comprising besides the cancer cells vasculature, extracellular matrix (ECM), soluble molecules and a plethora of various other cell types. These components form a so-called tumour microenvironment. From the numerous cell types that are part of tumor microenvironment, tumor infiltrating lymphocytes (TILs) play a major role in patient prognosis. Their presence is also of major importance with regard to new biological therapies based on immune checkpoint inhibitors. Crucial role of TILs is also reflected by the new approaches in cancer diagnostics namely by Immunoscore method (currently used in clinical settings). Immunoscore is based on localization and quantification of CD3+ and CD8+ TILs in thin histological sections of tumor tissue. The question remains to which extent the information obtained from 2D slices reflects the situation in tumor microenvironment considering its spatial heterogeneity. The development of new methodological approaches allowing evaluation of histological information in 3D is the key to answer this question. The theoretical part of this work first describes the heterogeneity of the tumor microenvironment and the role of immune cells within it. Then, the role of spatial heterogeneity and its possible influence on the histopathological...
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Exprese CD47 a jeho topologie na povrchu primárních buněk karcinomu močového měchýře při interakci s makrofágy
Rajtmajerová, Marie ; Drbal, Karel (advisor) ; Brdička, Tomáš (referee)
CD47 is a so-called "don't eat me" signal, which protects cells from phagocytosis. Its high expresion on tumor cells brings new perspective to the tumor therapy. Monoclonal antibodies, which are these days undergoing clinical trials, prevent CD47 binding to the SIRPA inhibitory receptor on macrophages, and so they enhance their phagocytic functional capacity. In this way they enable phagocytic removal of tumor cells. Overall expression, structural conformation and stoichiometry of CD47 on a particular cell predestine whether it will be phagocytised. The aim of the thesis is to develop and test methods to characterise expression parameters of CD47 via flow cytometry (FCM), quantitative PCR (qPCR) and microscopy. To achieve this goal I performed competition tests of commercially available antibodies in order to characterise their binding epitopes on cell lines. After performing tSNE analysis of primary BCa patient samples I correlated CD47 expression with other cell surface markers. I focused on CD47 expression in various differentiation stages of the tumor. To better understand the relationship between CD47 expression and differentiation status of cells I performed qPCR analysis of particular transcription factors. Using cell lines I examined method for phagocytosis quantification, which will be...
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Radiotherapy of cervical cancer
KUBIŠOVÁ, Lucie
This bacherol thesis focuses on radiotherapy of cervical cancer. In the theorical part we describe the anatomy proportions of little pelvis, typology of the tumor, its occurance, risk factors that can cause this disease, symptoms and diagnostics. We put special attention on the questions related to the prevention of cervical carcinoma. In addition we are interested in the treatment for this malignant disease surgery solutions, teletherapy, brachytherapy and chemotherapy. The second part of the bacherol thesis is empirical and focuses the radiotherapeutical treatment of patients with cervical carcinoma, stressing the analysis of doses of irradiation for the critical organs with repercussion for posterior occurance of acute or late undesired effects. The major risk factor detected was the insufficient participation of the respondents on the preventive gynecology examinations. The collected data reflect that only one third of women are undergoing the regular screening by their doctors. When analyzing the facts, we wanted to focuse on the irradiation doses applied to the critical organs. The obtained results showed that neither doses nor fractions have demostrated even a little progress during seven years. At the same time the techniques of irradiation did not evolve very much during the same period. The only difference was the increase of application of the IMRT technique in 2017 in comparison to 2010. The original hypothesis of the thesis advocating that the improvement of verification systems, imaging machines and planning systems could have an impact on the reduction of undesired effects caused by irradiation of critical organs during radiotherapy of cervix, was rejected due to above mentioned results.
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Immune response to experimental active immunotherapy DCVac/OvCa in patients with ovarian carcinoma in phase II clinical trials.
Ksandrová, Marie ; Sadílková, Lenka (advisor) ; Krulová, Magdaléna (referee)
The immunotherapeutic drug DCVAC/OvCa is being tested in the treatment of ovarian cancer patients within the SOV02 clinical trial (Eudra CT number: 2013-001323-38). Ovarian cancer belongs to gynaecological malignancies with the highest mortality rate. Around 60% of patients are diagnosed at advanced stages. Despite the initial successful treatment, relapses occur in most cases, and the disease often becomes resistant to chemotherapy. Effective therapy for relapsed or metastatic patients is still missing. The solution could be immunotherapeutic treatment. DCVAC is an active cellular immunotherapy based on autologous dendritic cells. The aim of this diploma thesis was monitoring of immune parameters in samples from clinical trial SOV02 patients during the time period defined in the study protocol. We have monitored the presence of antigen specific T lymphocytes, tumor specific antibodies, immunosuppressive populations of regulatory T cells and MDSC cells, and also the expression of inhibitory molecules on the surface of T lymphocytes. We observed higher levels of Her-2, Muc-1 and MAGE-A1 antibodies in the DCVAC/OvCa treated group of patients versus the control group. Significant differences in the other monitored parameters were not observed. However, a large amount of data have been obtained that...
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Surface phenotype of human carcinoma cancer stem cells (CSC)
Bočková, Marie ; Drbal, Karel (advisor) ; Čermák, Vladimír (referee)
Tumor is composed of a heterogenous mass of cells. Similar to normal healthy organs and tissues, these can be divided into individual cellular subpopulations according to morphology, function and expression patterns. A subpopulation of cells that are able to give rise to all of these cellular lineages is referred to as cancer stem cells (CSC). CSCs have the capabilities of normal stem cells such as the self-renewal and the ability to give rise to a heterogenous population of differentiated cells. Usually, this is the most resistant subpopulation within a tumor, highly non-responsive to therapy. Doing so, they are the cause of residual disease. Characterisation of CSC markers of individual tumor types is beneficial since it enables higher therapy efficacy via targeting this cell population. The -omics approaches to characterisation of the surface proteome bring a broader view into the field when searching for a unique gene signature of specific cancer stem cell types. It has been found that these cells can be identified based on the high expression levels of CD44, CD90 and CD49f. Among other markers, CD47 is an important marker for its immunosuppressive function.
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Thyroid tumors and their molecular genetic causes.
Šmídová, Barbora ; Dvořáková, Šárka (advisor) ; Soták, Matúš (referee)
The aim of this work is to summarize actual literature overview and to collect the up- to-date knowledge on genetic causes of the development of thyroid carcinomas. Thyroid carcinomas represent the most often endocrine malignancy and its incidence is still growing. This work describes all types of thyroid carcinomas derived from different cell types and are distinguished also according to the degree of differentiation. The main genetic changes in the tumor tissues of the medullary, papillary, follicular and anaplastic thyroid carcinoma are described. Thyroid carcinomas occur mostly in sporadic form, rarely as a familial disease. The causes of familial forms of thyroid carcinomas are also summarized and the main mutations in the germ-line DNA are identified. Key words: carcinoma, genetics, mutation, thyroid
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Thyroid tumors and their molecular genetic causes.
Šmídová, Barbora ; Dvořáková, Šárka (advisor) ; Koudelková, Lenka (referee)
The aim of this work is to summarize actual literature overview and to collect the up- to-date knowledge on genetic causes of the development of thyroid carcinomas. Thyroid carcinomas represent the most often endocrine malignancy and its incidence is still growing. This work describes all types of thyroid carcinomas derived from different cell types and are distinguished also according to the degree of differentiation. The main genetic changes in the tumor tissues of the medullary, papillary, follicular and anaplastic thyroid carcinoma are described. Thyroid carcinomas occur mostly in sporadic form, rarely as a familial disease. The causes of familial forms of thyroid carcinomas are also summarized and the main mutations in the germ-line DNA are identified. Key words: carcinoma, genetics, mutation, thyroid
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Study of genetic factors modifying the risk of onset and progression of colorectal and pancreatic cancer
Mohelníková Duchoňová, Beatrice ; Souček, Pavel (advisor) ; Skálová, Lenka (referee) ; Macek, Milan (referee)
Introduction: The aim of this study was to evaluate the role of genetic and lifestyle factors in the risk of onset and progression of colorectal and pancreatic cancer. The first part deals with the etiological factors and the importance of polymorphisms in biotransformation enzymes and genetic alterations in the gene CHEK2 in the origin of these malignancies. In the second part, the ABC transporter genes were analyzed as potential prognostic and predictive markers of a treatment's outcome. Materials and methods: The polymorphisms and other genetic alterations were detected using real-time PCR, allelespecific PCR and PCR-RFLP methods in DNA which was extracted from the blood of patients. The frequency of polymorphisms was evaluated and their importance was assessed with regard to the available epidemiological data. Gene expressions were determined by qPCR in paired samples of tumor tissue and adjacent non-tumorous parenchyma. Results: A majority of the observed polymorphisms failed to show a relationship between their presence and the risk of any of these malignancies. CYP2A13 variant allele*7 coding inactive enzyme was found in 7 of 265 controls and in none of 235 pancreatic carcinoma patients. In contrast, GSTP1-codon 105 Val variant allele and GSTT1-null genotype were associated with an elevated...
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