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Non-small cell lung cancer immunotherapy based on dendritic cell vaccine pulsed with tumor cells killed by immunogenic cell death
Podzimková, Naďa ; Adkins, Irena (advisor) ; Ryška, Aleš (referee) ; Froňková, Eva (referee)
Lung carcinoma represents the leading cause of cancer mortality worldwide with dismal prognosis. Immunotherapy exploiting the patient's own immune system is booming along with dendritic cell (DC)-based immunotherapy in recent years. The use of DC-based vaccines for treatment of non-small cell lung cancer is, however, still limited with limited clinical efficacy despite the development of various techniques for the vaccine preparation. In this study, we focused on the development and characterization of DC-based vaccine pulsed with multi-antigenic tumor cells killed with high hydrostatic pressure (HHP) and cytotoxic hyperthermia (cHT). Both physical modalities induced immunogenic cell death which leads to DC activation and more efficient onset of antitumor immunity. The molecular apoptotic pathways activated upon HHP and cHT treatment in tumor cells were analyzed and the optimal conditions to achieve high mortality, high immunogenicity and low antigen degradation were identified. We also showed that the prophylactic vaccination of mice with cHT-treated tumor cells significantly delayed tumor growth and prolonged mice survival. DC pulsed with HHP or cHT-treated tumor cells exhibited phenotypic maturation, increased chemotactic migration, higher production of proinflammatory cytokines and increased...
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Mechanisms of immune dysregulation leading to inflammatory bowel disease
Horáčková, Klára ; Froňková, Eva (advisor) ; Filipp, Dominik (referee)
Bc. Klára Horáčková DIPLOMA THESIS Mechanisms of immune dysregulation leading to inflammatory bowel disease Abstract Inflammatory bowel disease (IBD) is a complex disorder characterized by chronic inflammation of the gastrointestinal tract. Classical IBD is a multifactorial disease with adulthood or later-childhood onset. However, children with very early onset IBD (VEO-IBD, before 6 years of age) are a specific cohort, whose pathology can be caused by severe genetic defects in genes connected to immune homeostasis in the gut. We aimed to identify the causal genetic variants in 20 pediatric patients diagnosed with IBD (age of onset from 3 to 154 months) using whole exome sequencing (WES). We evaluated several bioinformatical approaches for WES data analysis. This included a comparison of two methods of variant identification using VarScan2 or GATK4-based tools. Furthermore, we compared 4 gene lists ("virtual panels") for variant filtering, one of which was compiled purposefully for this thesis. We identified and validated via segregation analysis 5 causal variants in 4 genes (DUOX2 compound heterozygote, FOXP3, NLRP3 and NOD2) accounting for 20 % of the cohort. NOD2 (p.A755V) variant has already been reported in IBD cases, while DUOX2 (p.R1216W + p.A1131T), FOXP3 (p.H400L) and NLRP3 (p.V200M) were newly...
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A study of interactions of novel anticancer drugs with drug transporters.
Šorf, Aleš ; Čečková, Martina (advisor) ; Kollár, Peter (referee) ; Froňková, Eva (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Mgr. Aleš Šorf Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Title of doctoral thesis: A study of interactions of novel anticancer drugs with drug transporters. Despite the significant progress in recent years, the efficiency of anticancer therapy is still complicated by drug resistance. This phenomenon can be mediated by pharmacokinetic mechanisms such as efflux via ATP-binding cassette (ABC) transporters and biodegradation of parent drug to inactive metabolites via metabolizing enzymes. These proteins also play a key role in absorption, distribution and excretion of drugs. When two or more interacting substances are administered simultaneously, the pharmacokinetic drug-drug interactions may occur. Since the targeted protein kinase inhibitors are becoming more common and important component of modern anticancer therapy, the aim of this study was to elucidate interactions of selected drugs of this novel therapeutic group with ABC transporters and also drug metabolizing enzymes by in vitro approaches. We further examined relevant substances ex vivo using mononuclear cells isolated directly from the blood of patients de novo diagnosed for acute myeloid leukemia (AML). Regarding the...
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A study of interactions of novel anticancer drugs with drug transporters.
Šorf, Aleš ; Čečková, Martina (advisor) ; Kollár, Peter (referee) ; Froňková, Eva (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Mgr. Aleš Šorf Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Title of doctoral thesis: A study of interactions of novel anticancer drugs with drug transporters. Despite the significant progress in recent years, the efficiency of anticancer therapy is still complicated by drug resistance. This phenomenon can be mediated by pharmacokinetic mechanisms such as efflux via ATP-binding cassette (ABC) transporters and biodegradation of parent drug to inactive metabolites via metabolizing enzymes. These proteins also play a key role in absorption, distribution and excretion of drugs. When two or more interacting substances are administered simultaneously, the pharmacokinetic drug-drug interactions may occur. Since the targeted protein kinase inhibitors are becoming more common and important component of modern anticancer therapy, the aim of this study was to elucidate interactions of selected drugs of this novel therapeutic group with ABC transporters and also drug metabolizing enzymes by in vitro approaches. We further examined relevant substances ex vivo using mononuclear cells isolated directly from the blood of patients de novo diagnosed for acute myeloid leukemia (AML). Regarding the...
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The aberration of chromosome 7 in haematological malignancies of the myeloid lineage
Onderková, Martina ; Ransdorfová, Šárka (advisor) ; Froňková, Eva (referee)
Accurate localization of breakpoints and deleted regions on chromosome 7 in bone marrow cells of patients is an essential step in identifying genes involved in tumor transformation of a cell. In case of hematological malignancies usually oncogenes and tumor suppressor genes are activated or deleted by a change in the arrangement of genetic material. Aberration of chromosome 7, total or partial loss of chromosome, especially long arms 7q, are among the recurrent cytogenetic abnormalities in patients with myeloid diseases such as myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). Aberrations of chromosome 7 are an important prognostic marker occurring in 8-10% de novo MDS and AML and in 40-50% treated MDS / AML. For a detailed analysis of chromosome 7 breakpoints and aberrations, samples of 51 adult patients diagnosed with AML / MDS were examined using conventional and molecular cytogenomic methods. In our testing group we demonstrated a separate 7q deletion in one patient (2%) and an isolated monosomy of chromosome 7 in six patients (12%). Aberration of chromosome 7 detected in combination with another change was found in 17 cases (33%) and 27 patients (53%) had complex karyotype changes including chromosome 7. The most frequent breakpoint was 7q22. In 26 patients we proved a deletion...
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Experimental therapy of B-cell Non-Hodgkin's lymphonas.
Klánová, Magdalena ; Klener, Pavel (advisor) ; Machová Poláková, Kateřina (referee) ; Froňková, Eva (referee)
1 ABSTRACT B-cell non-Hodgkin lymphomas (B-NHL) represent the most common mature lymphoproliferative diseases. B-NHL arise at different stages of B-cell development and represent their malignant counterpart. Diffuse large B-cell lymphoma (DLBCL) and mantle cell lymphoma (MCL) are aggressive types of B-NHLs. Deregulation of cell cycle control, inhibition of apoptosis or abnormal DNA damage response play a key role in the pathogenesis of DLBCL and MCL. Aberrant activation of several signaling pathways that further promote survival, cell proliferation or affect the tumor microenvironment have been recently recognized. Increased understanding of the oncogenic mechanisms implicated in pathogenesis of B-NHL lead to development of novel agents that target the oncogenic drivers of distinct lymphoma subtypes. MCL is an aggressive subtype of B-NHL associated with poor prognosis. In vivo models of human MCL for experimental therapy are however scarce. We established and characterized several mouse models of human MCL by xenotransplantation of either primary cells or established cell lines into immunodeficient mice (publication no 1). We demonstrated that engrafted MCL cells displayed complex changes of gene expression profile, phenotype and sensitivity to cytotoxic agents compared to the original in vitro growing...
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Study of HLA antigens and KIR genes in a donors and recipients of bone marrow
Kroulíková, Zuzana ; Vraná, Milena (advisor) ; Froňková, Eva (referee)
HLA and KIR genes are highly polymorphic regions within the human genome. Protein products of these genes play a critical role in hematopoietic stem cell transplantation. Genetic HLA match is a major barrier to engraftment and influences the outcome of this therapy. Therefore it is necessary to genotype donors and recipients selected for hematopoietic stem cell transplantation. Today HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 genes are tested by modifications of polymerase chain reaction or by sequence-based typing methods on the level of high- or low-resolution. Donors registered in bone marrow registries are selected on the basis of a 10/10 match. Donors KIR genotype leads to a better outcome, to relapse-free survival and overall survival in treatment of patients with acute myeloid leukemia. A better protection against relapse is achieved by Cen-B/B donor haplotype. Therefore KIR typing by polymerase chain reaction is used and the genotype is compared with the IMGT/KIR database by an online B- content calculator. Donors are divided in groups according to their genotype and their influence on the success of treatment for acute myeloid leukemia. The study of polymorphic systems and the development of genotyping donors and recipients significantly improve the outcome of hematopoietic stem cell...
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