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Induction of beta-cell proliferation by synthetic modified mRNAs encoding cell cycle regulators
Ivanovská, Dana ; Koblas, Tomáš (advisor) ; Černá, Věra (referee)
Diabetes mellitus is a metabolic disease associated with a high blood glucose level over a prolonged period of time. Hyperglycemia is caused by the loss of pancreatic insulin producing beta cells. Diabetes mellitus II is linked with insulin resistence, which can indirectly lead to beta cell deficiency. It logically follows that the replacement or regeneration of beta cells could lead to a successful remission of diabetes. D type cyclins (D1, D2, D3) and cyclin-dependent kinases (Cdk) 4/6 appear to have the potential to induce beta cell proliferation. These proteins are responsible for driving cell mitotic entry. The aim of this bachelor thesis was to verify the possibility of inducing beta cell proliferation via D type and Cdk4/6 synthetic mRNA transfection. In vitro-synthesized mRNA induces short-therm protein overexpression. Cyclins harboring mutations are characterized by a higher protein stability and an increased half-life. The presence of D type cyclins and Cdk4/6 after cell transfection was detected using indirect immunofluorescence. Also a Western blot analysis with subsequent immunodetection was performed. Transfecting rat islet cells with various D type cyclins and Cdk4/6 mRNA combinations has shown to lead to a significant induction of beta cell proliferation. The levels of beta cell...
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Induction of beta-cell proliferation by synthetic modified mRNAs encoding cell cycle regulators
Ivanovská, Dana ; Koblas, Tomáš (advisor) ; Černá, Věra (referee)
Diabetes mellitus is a metabolic disease associated with a high blood glucose level over a prolonged period of time. Hyperglycemia is caused by the loss of pancreatic insulin producing beta cells. Diabetes mellitus II is linked with insulin resistence, which can indirectly lead to beta cell deficiency. It logically follows that the replacement or regeneration of beta cells could lead to a successful remission of diabetes. D type cyclins (D1, D2, D3) and cyclin-dependent kinases (Cdk) 4/6 appear to have the potential to induce beta cell proliferation. These proteins are responsible for driving cell mitotic entry. The aim of this bachelor thesis was to verify the possibility of inducing beta cell proliferation via D type and Cdk4/6 synthetic mRNA transfection. In vitro-synthesized mRNA induces short-therm protein overexpression. Cyclins harboring mutations are characterized by a higher protein stability and an increased half-life. The presence of D type cyclins and Cdk4/6 after cell transfection was detected using indirect immunofluorescence. Also a Western blot analysis with subsequent immunodetection was performed. Transfecting rat islet cells with various D type cyclins and Cdk4/6 mRNA combinations has shown to lead to a significant induction of beta cell proliferation. The levels of beta cell...
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Possibilities of prediction and immunointervention in type 1 diabetes
Sklenářová, Jana ; Štechová, Kateřina (advisor) ; Saudek, František (referee) ; Pavlínková, Gabriela (referee)
Type 1 diabetes mellitus (T1D) is an organ-specific autoimmune disease characterised by autoimmune destruction of insulin-producing beta cells in the islets of Langerhans. It is a long-term process initiated months or even years prior to the clinical onset. The main role in the pathogenesis is played by T lymphocytes but other cell types are involved as well. The presence of autoantibodies in the circulation is typical even before the disease onset. Nowadays, intensive research is focused on finding individuals at risk and developing an effective prevention. During my postgraduate studies I was involved mainly in the research of T1D prediction and prevention. We investigated the relationship of established autoimmune markers - autoantibodies - and the cellular reactivity to GAD65 and IA2 autoantigens. We discovered that the reaction to autoantigens is very individual and it is influenced by the patient's autoantibody profile. These results could be relevant in planning antigen-specific immunointervention studies and improving their efficacy. We also made an attempt to improve specificity and sensitivity of a beta cell destruction marker (specifically demethylated DNA), which would enable better understanding of the beta cell decline and identification of individuals at risk of T1D development. In...
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