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Monitoring of immune parameters during anti-tumor immunotherapy
Bílková, Pavla ; Palich Fučíková, Jitka (advisor) ; Fialová, Anna (referee)
Dendritic cells are the most effective antigen presenting cells in humans, they stimulate naive T lymphocytes and thus initiate specific immune response. The discovery of dendritic cells and understanding of their functions contributed to the idea of usingdendritic cells for the treatment of cancer. Anti tumor immunotherapy is a therapeutic strategy that aims to induce and maintain immune responses against tumor cells. Currently, immunotherapy based on dendritic cells has strong position among other anti cancer therapies and seems to be a promising therapeutic option for patients with tumors. In this work, I evaluated the effectiveness of treatment in patients with prostate cancer treated with immunotherapy based on dendritic cells. I focused on the detection of antigen specific T lymphocytes in peripheral blood against tumor antigens, PSA, NY ESO 1, MAGE A1 and MAGE A3. Using a 3 day standard protocol for the detection of antigen specific T cells using intracellular cytokine staining we were able to detect only a small percentage of this minor population. Only after extension of the protocol, we increased the sensitivity setting and we detected a significantly increased frequency of antigen specific T lymphocytes in the peripheral blood after one year DC vaccines application.
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Prostate Cancer Diagnostic Algorithm
Sedláčková, Hana ; Hora, Milan (advisor) ; Baxa, Jan (referee) ; Záleský, Miroslav (referee)
Prostate cancer diagnostic algorithm Aim: The aim of the study is to implement the latest scientific knowledge in the diagnosis of prostate cancer (PC). We focused on tumor markers, imaging methods, prostate biopsy methodology and we created a diagnostic algorithm based on a review of current literature in combination with our own experience. Material and methods: The algorithm is divided into several branches, which have been individually subjected to clinical studies. Due to the low sensitivity and specificity of PSA, prostate health index (PHI) was added to the first line of patient stratification. 787 patients were primarily examined and these subsequently underwent radical prostatectomy. PHI levels were compared with definitive staging and grading. Cut-off values for PC detection and high-risk stratification, including locally advanced PC were determined. Next, 320 patients underwent prostate biopsy followed by radical prostatectomy. The cohort was further divided into two subgroups, patients with GS = 6 and patients with GS > 6. The ability of PHI to distinguish between insignificant and significant prostate cancer was evaluated. In a multicentric study with 395 patients, PHI with additional markers (tPSA, PSAD) and multiparametric magnetic resonance imaging of prostate (mpMRI) was assessed....
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Molecular mechanisms responsible for regulation of apoptosis in prostate cancer cells treated with TRAIL and chemotherapeutic drugs
Tománková, Silvie ; Hyršlová Vaculová, Alena (advisor) ; Špegárová, Jarmila (referee)
Prostate cancer is one of the most common cancer-related causes of death among men. Chemotherapy is mainly used for treatment of its later stages, accompanied by unpleasant side effects. So far, the treatment of advanced stages of prostate cancer has not been sufficient, and new more effective alternatives are needed. The application of the TRAIL cytokine, which induces apoptosis in tumor cell, but is not toxic to nonmalignant cells, seems to be a promissing approach. However, TRAIL-based therapy is often limited by the emerging cancer cell resistance. Overcoming the resistance can be achieved by combination therapy of TRAIL with effective sensitizers. Within this work, a combination of TRAIL with platinum-based chemotherapeutic drugs such as cisplatin or its novel derivative LA-12 was applied in order to facilitate the elimination of prostate cancer cells. In the experimental part of this work, using Western blot and flow cytometry analysis it was shown that TRAIL in combination with CDDP or LA-12 effectively enhanced apoptosis in three human prostate cancer cell lines. This effect was accompanied with increased activation/amount of several proapoptotic Bcl-2 family proteins, while no changes in the level of the receptors for TRAIL were observed. These results demonstrated that especially the combination...
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The use of immunochemical methods in the diagnostic of cancer and degenerative disease
Hanousková, Lenka ; Kotaška, Karel (advisor) ; Bronský, Jiří (referee) ; Štern, Petr (referee)
Introduction: Immunochemical methods have significant importance in biomedical research. The benefit of immunochemical methods is their high sensitivity and accuracy. The aim of the work was to use immunochemical methods in investigation of cancer biomarkers and degenerative diseases. A total of four studies were performed. Two studies were focused on the search for new biomarkers of prostate cancer, the third study was focused on evaluation of FGF23 in patients with hyperparathyroidism, and the fourth study looked at the assessment of hyaluronic acid in synovial fluid of patients with knee gonarthrosis. Methods and patients: Study 1: Concentrations of Chromogranin A, Endoglin, TIMP-1 and thymidine kinase 1 (TK1) in serum and Endoglin, SPINK-1, Annexin, TIMP-1 in urine were measured in 58 patients with the (mean age 68 years, range 45 - 82 years) with clinically diagnosed prostate cancer and in 30 healthy individuals with the (mean age 64 years, range: 55 - 78 years) without clinically relevant urological history and PSA values in the reference range. Subsequently, serum TK1 levels were measured in 169 patients (mean age 62 years, range 45- 82 years) with prostate cancer and in a control group of 39 healthy men (mean age 61 years, range 54-78 years). The TK1 assay was also used to determine the...
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Perineural spread of pelvic tumors: mechanism and diagnosis
Čapek, Štěpán ; Sameš, Martin (advisor) ; Haninec, Pavel (referee) ; Hořínek, Daniel (referee)
Perineural spread of pelvic tumors mechanism and diagnosis Abstract Neoplastic lumbosacral plexopathies are infrequent affections of the lumbosacral plexus. Cases with minimal or non-specific finding on imaging can be particularly puzzling to diagnose. We describe a series of patients with perineural spread from the site of the primary tumor along the visceral autonomous nerves to the lumbosacral plexus and further proximally to the spinal nerves and even intradurally and also distally to the sciatic nerve. On series of 17 patients diagnosed with perineural spread of pelvic malignancy we describe characteristic clinical presentation and imaging finding. In many of these cases a tissue biopsy is necessary to finalize the diagnosis. We describe operative technique of targeted fascicular biopsy of the sciatic nerve and our experience with this procedure. On series of 117 patients, we report the outcome and complication: diagnostic yield was 84.8% and complication rate was 2.7 %. If a tissue sample is needed to conclude the diagnosis, targeted fascicular biopsy does increase the yield at an acceptable complication rate. Perineural spread of pelvic cancer is a new clinical-pathological entity with an unknown natural history or ideal treatment strategy. Based on the imaging finding in this group we present a...
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Adoptive transfer of tumor-specific lymphocytes for cancer immunotherapy
Vávrová, Kateřina ; Bartůňková, Jiřina (advisor) ; Němečková, Šárka (referee) ; Reiniš, Milan (referee)
Prostate cancer is the second leading cause of cancer death in men in Europe and the US. In the context of previous preclinical experiments and clinical studies there are certain assumptions predicating successful application of immunotherapy in the treatment of patients with prostate cancer. Promising results have been achieved by a combination of different treatment modalities which provide a synergistic antitumor effect. One of these combinatorial options is the use of antitumor vaccines and adoptive T cell transfer. The topic of this thesis is to provide a fresh insight into the past and current trends following the long-term candidate's department program in the field of anti-tumor immunotherapy. The experimental part of this thesis revolves around our own results published in this field. The introductory chapter delivers a basic overview of cellular mechanisms of anti-tumor immunity and the role of individual immune components in these processes. Following chapters are dedicated to current immunotherapeutic approaches with emphasis on the adoptive T cell transfer and implication of this technology in the treatment of prostate cancer. The results section describes the establishment of our protocol for adoptive T cell transfer as well as the protocol for ex vivo enrichment of human T cell...
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Verification of target volume position in radioterapy
LHOTÁKOVÁ, Vendula
This Bachelor thesis is concerned with the verification of the target volume setup in radiotherapy. With respect to technological advancement, the necessity to use verification systems for ensuring a quality treatment is highlighted. In its theoretical part, the objective of this Bachelor thesis is to summarize and delineate the imaging technologies used to verify the target volume setup. The objective of the practical part is to analyse deviations in the position of the target volume in IMRT technique for prostate radiation; the analysis is primarily directed at new functions in the correction of the position of the radiation table. The theoretical part of the thesis presents facilities used to verify the target volume and summarizes information relating to the two basic methods in radiotherapy: teletherapy and brachytherapy. Differences between X-ray and radionuclide irradiators and particle accelerators are explained. A brief description is given as to the structure of a linear accelerator, with a summary of irradiation techniques applied in radiotherapy. The data for the practical part were derived from results gained by the correction of the position of the target volume in verification imaging at the Radiotherapy Department in České Budějovice Hospital. For the analysis of deviations, a set of 50 patients who were treated for prostate cancer at the Radiotherapy Department in 2018 was selected. During radiotherapy, excepting daily MV-kV imaging, the total of 417 verification CTs was developed for these subjects. Based on these CT images, the deviation in directing the target volume was evaluated and the appropriate correction of the radiation table, and the given patient as well, was performed. The correction of the radiation table allows for the movement of six axes in total, as described in detail in the present thesis. The results gained from the data collected reveal that the correction of the setup of the irradiated volume is a necessity and it is amply used, specifically in all of the six axes. It was discovered that the functions of the table Lat, Lng, Rnt and Vrt are used at virtually all times, and in concrete terms, the newer function of the table Pitch is utilized at 95,03%, with the use of function Roll standing at 92,64%. The daily verification CBCT was performed in 10% of the patients in the set observed. The standard frequency of the use of the verification CBCT at the Radiotherapy Department in České Budějovice Hospital, for patients irradiated to prostate, is approximately 7 - 9 CBCT during the treatment, which includes a CBCT examination at the first setup and then twice during the first week of treatment, and subsequently only once a week during the radiotherapy procedure. The analysis results proved an abundant use of newly introduced options in the correction of the position of the radiation table, and in general, also an improvement in accuracy of directing the target volume. The improvement in accuracy, and therefore an improvement of quality of the administered radiation therapy, allows for changes in the size of the recorded target volumes, specifically the reduction in these volumes. As a consequence, it is logical that the volume of healthy irradiated issue is smaller, which brings (or might bring) for the patient a decrease in the occurrence or level of seriousness of undesired radiotherapy effects. The results of the research and its findings may serve for evaluating the quality of the verification process in radiotherapy, linked with installation of new technologies in the given healthcare department, or, as the case may be, as data and information to be used for preparation of articles published in specialized magazines.
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Detection of circulating tumour cells and their clinical application in patients with bioptically proven prostate cancer.
Čapoun, Otakar ; Soukup, Viktor (advisor) ; Zachoval, Roman (referee) ; Veselý, Štěpán (referee)
1 ABSTRACT Introduction and aim of the study Circulating tumor cells (CTCs) are a promising tool of identifying patients with castration- resistant prostate cancer (CRPC) who will benefit from often demanding cytotoxic therapy. The aim of this work was to evaluate the prognostic significance of CTC in docetaxel-treated CRPC patients. During the project, we also tested the various methods of CTC cultivation and studied their genetic profile as well as the genetic profile of histological specimen at the time of diagnosis. Patients and methods A total of 39 patients who met the CRPC criteria and were indicated for docetaxel chemotherapy were included in the prospective study. Blood collection for CTC analysis was done in all patients before chemotherapy and on the first day of the fourth or fifth cycle of docetaxel. In parallel, CTCs were cultivated. Isolation and detection of CTC was done using the AdnaTest system, which consists of immunomagnetic separation and subsequent detection of mRNA from the CTC lysate. The primary objective of the study was to evaluate the overall survival (OS) of patients. Survival analysis was performed using the Kaplan-Meier method of estimating the survival distribution function. The impact of individual factors was tested using the Log-rank test, the Wilcoxon test and the Cox...
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Nanodiamonds as an innovative system for intracellular delivery of mirna-34a inprostatic cancer therapy
Bitti, G. ; Abate, M. ; Neuhoferová, Eva ; Kindermann, Marek ; Petráková, V. ; Boccellino, M. ; Quagliuolo, L. ; Filová, Eva ; Benson, Veronika ; Caraglia, M. ; Amler, Evžen
The microRNA(miRNA)-34a is an important regulator of tumor suppression. It controls the expression of several target proteins involved in cell cycle, differentiation and apoptosis, and antagonizes processes that are necessary for basic cancer cell viability as well as cancer stemness, metastasis, and chemoresistance. It is downregulated in numerous cancer types, including prostatic cancer, and inhibits malignant growth by repressing genes involved in various oncogenic signaling pathways. Given the anti-oncogenic activity of miR-34a, here we proved the substantial benefits of a new therapeutic concept based on nanotechnology delivery of miRNA mimics. In order to monitor the miRNA-34a replacement, we used a fluorescent nanodiamond particles (FND) system with linked miRNA-34a mimic, which was delivered to PC3 and DU145 prostatic cancer cell lines. We used functionalized nanodiamonds coated with polyethylenimine to transfer miRNA-34a into PC3 and DU145 prostatic cancer cell lines and we measured the zeta-potential of these complexes before using them for in vitro experiments. A replacement of miRNA-34 was observed by monitoring levels of miRNA-34 via real-time PCR. Moreover, our in vitro experiments demonstrated that miRNA-34a replacement, using this FND delivery system, decreased viability and induced apoptosis in prostatic cancer cell lines. Our findings suggest the replacement of oncosuppressor miRNA-34a provides an effective strategy for cancer therapy and the FND-based delivery systems seems to be an excellent strategy for a safe and effective targeting of the tumor.
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Study of the mechanism of anticancer drug action on neuroblastomas
Černá, Tereza ; Stiborová, Marie (advisor) ; Souček, Pavel (referee) ; Mrízová, Iveta (referee)
Despite advances in cancer diagnosis and therapy, cancer is the second leading cause of death globally. The improvements of cancer treatment are the major challenge in this research. The aim of the thesis was studying of effects of two anticancer drugs ellipticine (Elli) and doxorubicin (DOX) on some cancer and healthy cell lines. Specific consideration was given to expand current knowledge about the metabolism and cytostatic effects of Elli in neuroblastoma cell lines. Another part of this study was focused on mechanisms contributing to the development of ellipticine-resistance in cancer cells and influence of histone deacetylase inhibitors on anticancer therapy was investigated. Moreover, the aim was to develop apoferritin (Apo) nanocarrier suitable for the active transport of cytostatics to cancer cells. Several essential data were found in this doctoral thesis. Anticancer efficiency of Elli depends on the CYP3A4-mediated metabolism in cancer. The CYP3A4 enzyme encapsulated into two nanoparticle forms, liposomes and SupersomesTM , was tested to activate ellipticine to its reactive species forming covalent DNA adducts. The formation of adducts seems to be dependent on concentrations of CYP3A4 in nanoparticle systems. A higher effectiveness of CYP3A4 in SupersomesTM than in liposomes to form...
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