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Role of tissue-specific expression of the ISWI protein Smarca5/Snf2h in murine hematopoiesis.
Turková, Tereza ; Stopka, Tomáš (advisor) ; Forejt, Jiří (referee) ; Mráz, Marek (referee)
The process of cell differentiation is dependent on the chromatin-remodeling activity of the ISWI ATPase SMARCA5 (SNF2H) and its complexes. A series of mouse models have been used to study the functions of this ATPase, starting with a model with constitutive deletion of the Smarca5 gene and followed by models with tissue-specific deletion of Smarca5 gene (especially at different stages of hematopoiesis). The severity of phenotypic manifestations in mouse models varies from early embryonic lethality in the constitutive deletion model, through fetal lethality associated with erythropoiesis failure (Vav1iCre model - deletion in the definitive hematopoietic progenitor) to lymphopoiesis defects that do not limit animal survival (hCD2iCre model - deletion in T and B lymphocytes) in conditional deletion models. This work builds on the observations obtained in the study of deletion models of this Smarca5 gene, i.e. that deletion leads to arrest of cell proliferation and their entry into apoptosis. Defects were observed already at the stem cell and hematopoietic progenitor stages, which were unable to enter the differentiation process in the absence of the SMARCA5 protein. In these models, it was not possible to study the progression of differentiation because of this reason. Therefore, we decided to create...
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CRISPR/Cas9 editing of leukemic B-cells: searching for microRNA-155 targets involved in the process of leukemogenesis
Sypecká, Markéta ; Savvulidi Vargová, Karina (advisor) ; Mráz, Marek (referee)
Markéta Sypecká CRISPR/Cas9 editing of leukemic B-cells: searching for microRNA-155 targets involved in the process of leukemogenesis Introduction: Chronic lymphocytic leukemia (chronic lymphoid leukemia, CLL) is a monoclonal disorder characterized by a progressive accumulation of functionally incompetent lymphocytes. CLL is the most common form of leukemia found in adults in Western countries. Course of the disease can differ: some patients die rapidly, within 2-3 years of diagnosis, because of complications from CLL, but most patients live 5-10 years. However, every stage of this disease has significantly higher level of miR-155, which is known as oncomiR. Micro RNAs represent negative regulators of gene expression. MiR-155 affects genes, which are involved in leukemogenesis and cell cycle. And it is known, that miR-155 suppresses its targets. We hypothesized that by gene editing of CLL B - cells we unblock miR-155 targets and find out correlation between these targets (known and unknown) with CLL leukemogenesis. Method we use for gene editing is CRISPR/Cas9, which enables to delete sequence of mature miR-155 in genome of leukemic B-cells. Methods: CRISPR/Cas9, nucleofection, qRT-PCR, FACS Results:We achieved to isolate clone that bears one allelic deletion (miR-155-/+) in sequence for mature...
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Up-regulation of microRNA miR-155 is reflected by low levels of its target mRNA encoding transcription factor PU.1 in primary tumors of human lymphomas
Hušková, Hana ; Stopka, Tomáš (advisor) ; Mráz, Marek (referee)
Lymphomas are heterogenous class of diseases characterized by proliferation of a malignant lymphocyte clone. MicroRNA miR-155 was found to be a key molecule in immune response, namely in inflammation and germinal reaction of B cells. On the other hand, miR-155 can drive lymphoproliferation in mouse and its levels were found to be elevated in certain lymphoma types in human. MiR-155 down-regulates expression of its target gene PU.1, a hematopoietic transcription factor important for B cell differentiation. Expression of the gene encoding miR-155, known as MIR155HG, is controled by several transcription factors, among them MYB, a member of an oncogenic E-box protein family. Levels of MYB itself are controled by microRNA miR-150. In this study, we measured levels of miR-155, PU.1, MYB and miR-150 in lymph nodes of patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CLL/SLL, N=20), diffuse large B-cell lymphoma (DLBCL, N=24), follicular lymphoma (FL, N=29), Hodgkin lymphoma (HL, N=25), marginal zone lymphoma (MZL, N=13), and mantle cell lymphoma (MCL, N=10). We also measured levels of these molecules in lymph nodes with the finding of strong inflammation (N=4). We found that patients of all the diagnoses except of MCL display heterogeneously elevated levels of miR-155 and correspondingly...
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Prevention of diabetes I. type and health promotion, education of diabetics and patients after pancreas transplantation
MRÁZ, Marek
The bachelor theses attend to people with diabetes mellitus Type 1 and patients after pancreas transplantation. The first chapters of theoretical part deal with pancreas anatomy, matter of disorder, its medication, belated complications, movement and nutrition connected with diabetes. The last chapter of this part is about pancreas transplantation. The practical part shows importance of education and knowledge of diabetics. It deals with life quality of diabetics who undergo pancreas transplantation. It compares their health and mental condition during diabetes, before and after pancreas transplantation.
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