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The expression profile of cardiovascular disease associated microRNAs in pregnancies with clinical manifestation of gestational hypertension, preeclampsia and intrauterine growth restriction
Bohatá, Jana ; Hromadníková, Ilona (advisor) ; Korabečná, Marie (referee)
MicroRNA (miRNA) are small non-coding 21-23 nucleotides long one strand RNAs. They are among the major posttranscriptional regulators of gene expression that regulate both physiological and pathological processes. Some of microRNAs, amount of their expression respectively, are specific only for certain type of tissue or pathological condition. The hypothesis for my diploma thesis was that gene expression of 28 cardiovascular disease associated microRNAs (miR-1-3p, miR-16-5p, miR-17-5p, miR- 20a-5p, miR-20b-5p, miR-21-5p, miR-23a-3p, miR-24-3p, miR-26a-5p, miR-29a-3p, miR-92a-3p, miR-100-5p, miR-103a-3p, miR-125b-5p, miR-126-3p, miR-130b-3p, miR-133a-3p, miR-143-3p, miR-145-5p, miR-146a-5p, miR-181-5p, miR-195-5p, miR- 199a-5p, miR-210-3p, miR-221-3p, miR-342-3p, miR-499a-5p, miR-574-3p) would differ in umbilical cord blood between groups of women with physiological pregnancies (FG), gestational hypertension (GH), preeclampsia (PE) and fetal growth restriciton (FGR). The studied cohort consisted of 184 pregnant women involving 44 controls, 47 GH pregnancies, 56 PE pregnancies and 37 FGR pregnancies. Relative quantification of microRNAs was performed by quantitative real-time PCR. Results showed a trend to miR-195-5p down-regulation in umbilical cord blood of GH patients. On the other hand, mild PE...
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The role of microRNA in hypoxic pulmonary hypertension
Tichý, Václav ; Hampl, Václav (advisor) ; Kolář, David (referee)
Pulmonary Arterial hypertension (PAH) is a devastating progressive disease that significantly decreases quality of life and has the average survival rate of only few years. One of the significant initiators of PAH is chronic hypoxia. After more than six decades of research that was initiated in 1946 by von Euler and Liljestrand, a new group of potential regulators of this pathology was discovered, that became heavily studied in the last five years. They are highly conserved molecules belonging to non-coding RNA. These 19-23 nucleotides long microRNA (miRNA) act as negative regulators of expression on various proteins. Many of them regulate traditional signalling pathways of hypoxic PAH (HIF-1, BMPR2) and miRNA is in turn regulated by other signalizations. Together, that creates an interconnected network of direct and indirect interactions and feedback loops, that we need to study in order to understand hypoxic PAH. This thesis summarizes findings about important miRNA molecules from the last few years and elucidates part of these regulatory mechanisms on several miRNA molecules (miR-17-92, miR-21, miR-210, miR-204 a miR143/145).
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Relation between cardiovascular microRNAs and pregnancy-related complications
Dalecká, Linda ; Hromadníková, Ilona (advisor) ; Daňková, Pavlína (referee)
During the pregnancy, there are many complications, which can affect a fetal development and eventually, its or mother's life. Pre-eclampsia is one of these complications. It is a hypertensive disorder, which appears during the pregnancy. Another example of these complications is an intrauterine growth restriction. It is a condition, when the fetus is not able to reach its genetical growth potential. Both of those pathological disorders are accompanied by changes in microRNA gene expression in placenta, for example miR-16, miR-21, miR-210. Knowledge about these changes in gene expression could represent unique instruments in the field of noninvasive prenatal diagnosis of these disorders. This could be benefitial due to the fact, that both of these disorders are responsible for increasing maternal and perinatal morbidity and mortality. This work focused on microRNAs, their expression in aformentioned disorders and benefits, which could bring in diagnostics. In this work these two disorders, characteristics of microRNAs and their biogenesi are described. MicroRNAs are interesting for us as potential biomarkers for noninvasive prenatal diagnostics becouse they are present in body fluids, such as plasma, breast milk or amniotic fluid and they are not subject of rapid degradation. Key words:...
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The role of biomarkers in erosive osteoarthritis of the hands
Lennerová, Tereza ; Šenolt, Ladislav (advisor) ; Tachezy, Ruth (referee)
Hand osteoarthritis (OA) is a degenerative joint disease that causes pain, functional limitation and negatively affects the patients' quality of life. The most severe subtype of this disease is erosive OA. Erosive hand OA is characterized by an abrupt onset, inflammation and is linked to worse outcomes than non-erosive hand OA. Current methods do not allow early diagnosis or to distinguish between patients with different forms at disease onset. This could be changed by the utilization of biomarkers in clinical practice. Biomarkers are molecules released into circulation that reflect biological processes. The main goal of this study was to analyze the levels of circulating biomarkers with the aim to differentiate patients from healthy subjects and patients with erosive OA from patients with non-erosive disease. Serum concentrations of seven biomarkers and the expression of plasma microRNAs were determined. Patients with hand OA showed altered cartilage metabolism, increased levels of adiponectin, decreased levels of clusterin and a dysregulated expression of several microRNAs in comparison to the healthy population. Patients with erosive OA had lower levels of clusterin and decreased expression of miR-151-3p than those with the non-erosive form of the disease. These findings suggest the potential...
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Vývoj chemických regulátorů drah mikroRNA a RNAi
Bruštíková, Kateřina ; Svoboda, Petr (advisor) ; Bařinka, Cyril (referee) ; Pospíšek, Martin (referee)
MicroRNAs are noncoding RNAs inducing sequence-specific posttranscriptional inhibition of gene expression and represent the major class of small endogenous RNAs in mammalian cells. Over 2,500 of human microRNAs potentially regulating more than 60% of human protein-coding genes have been identified. MicroRNAs participate in the majority of cellular processes, and their expression changes in various diseases, including cancer. Currently, there is no efficient small chemical compound available for the modulation of microRNA pathway activity. At the same time, small chemical compounds represent excellent tools for research of processes involving RNA silencing pathways, for biotechnological applications, and would have a considerable therapeutic potential. The presented work represents a part of a broader project, whose ultimate goal is: (i) to find a set of small molecules allowing for stimulation or inhibition of RNA silencing and (ii) to identify crosstalks between RNA silencing and other cellular pathways. This thesis summarizes results from the first two phases of the project, the development of high-throughput screening assays and the high- throughput screening (HTS) of available libraries of small compounds. To monitor the microRNA pathway activity, we developed and optimized one biochemical...
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CIrcadian regulation of miRNA and clock-controlled genes in tumorigenesis
Balounová, Kateřina ; Pácha, Jiří (advisor) ; Bendová, Zdeňka (referee)
The circadian clock generates circadian rhythms, which participate on regulation of a number of signalling pathways. Disruption of the circadian regulatory mechanism is linked to a development and a progression of certain types of cancer including colorectal tumorigenesis. Progression of tumorigenesis depends on the cell cycle machinery related to cell proliferation and apoptosis. MiRNAs play a role in initiation and progression of tumorigenesis because they interfere in regulatory pathways associated with tumorigenesis. The aim of the thesis was to determinate existence of circadian rhytms in clock controlled genes (Tef, Dbp), miRNAs (miR-1-3p, miR-16-5p, miR-34a-5p, miR-155-5p, miR-192-3p) and genes of the cell cycle machinery (Ccnd1, Ccne1, Ccna1, Ccnb1) and apoptosis (Casp3, Bcl2, Bad). Further, to compare detected circadian rhythms during aging and neoplastic transformation of colon by quantitative RT-PCR. We have observed circadian expression of Tef, Dbp, Ccne1, Ccna1, Ccnb1, Casp3 and Bcl2 in young mice colon, Tef, Dbp, miR-1-3p, Ccne1, Ccna1 in old mice colon and Tef and Dbp in colorectal tumors. In summary, circadian expression of clock controlled genes varied but was maintained in mice colorectal tumors. In aging we demonstrated weakening of circadian rhythms of the genes of the cell...
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Study of placental specific microRNA expression in pacients with spontaneous preterm birth and preterm prelabor rupture of membranes
Vintrová, Iva ; Hromadníková, Ilona (advisor) ; Daňková, Pavlína (referee)
MicroRNAs (miRNAs) are small non-coding RNAs with a length of 18 to 25 nucleotides playing a pivotal role in post-transcriptional regulation of gene expression. There are miRNAs whose expression is limited to a certain tissue type and diseases which are characterized by a unique miRNA expression profile. I assumed spontaneous preterm birth (PTB) and preterm prelabor rupture of membranes (PPROM) would be characterized by a unique miRNA expression profile. I observed the gene expression of 15 placental specific miRNAs (miR-512-5p, miR-515-5p, miR-516b-5p, miR-517-5p, miR-518b, miR-518f-5p, miR-519a-5p, miR-519d-3p, miR-519e- 5p, miR-520a-5p, miR-520h, miR-524-5p, miR-525-5p, miR-526a and miR-526b-5p) in placental tissue of pacients with PTB, PPROM and women with term in labor pregnancies (FG). PTB group consisted of 24 pacients, PPROM group of 75 pacients and FG group of 20 pacients. Quantitative real-time PCR was used to quantify gene expression. In the group of PTB pregnancies I identified 3 significantly upregulated miRNAs (miR-516b-5p, miR-519d-3p and miR-524-5p) and 4 miRNAs (miR-518b, miR-519a-5p, miR-520h and miR-526a) with a trend to upregulation compared to controls (FG). In the group of PPROM pregnancies I identified 3 miRNAs (miR-519d-3p, miR-520h and miR-256b-5p) with a trend to...
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Epigenetické regulace u autoimunitních onemocnění se zaměřením na revmatoidní artritidu
Horková, Veronika ; Daňková, Pavlína (advisor) ; Hušáková, Markéta (referee)
Exact cause of rheumatoid arthritis, as well as other autoimmune diseases has not been identified yet. In last twenty years, epigenetics showed a new face of immune system. DNA methylation, modification of histones - proteins around which DNA is wrapped, or interference of small RNA sequences - microRNAs, these all are heritable changes outside the DNA sequence that provide another component involved in autoimmunity. Presented epigenetic mechanisms alter gene expression and thus facilitate production of pro- inflammatory factors leading to autoimmune reactions. Moreover, genes regulating apoptosis are also frequently targeted by epigenetic modifications. Not only these mechanisms provide another level of immune defense, they also explain higher female susceptibility to autoimmune diseases and the influence of environment on pathogenesis of these diseases.
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Role of microRNA in regulations of circadian rhythms and tumorigenesis
Balounová, Kateřina ; Pácha, Jiří (advisor) ; Červená, Kateřina (referee)
MicroRNAs (miRNAs) are ~22nt long single-strand RNA found in all groups of organisms, where they affect biochemical, physiological and behavioral pathways by regulation of protein expression. Regulation of protein expression is mediated by silencing mRNA of target genes in one of two processes, translation repression or degradation of mRNA. Changed expression of miRNA can lead to aberrant regulatory pathways resulting in various pathophysiological conditions like cardiovascular diseases, cancer or neurological disorders. MiRNA can play a role in cancer both as an oncogen or as a tumor suppressor, and it is tissue and cancer-type specific. In colorectal cancer miRNAs downregulate or upregulate signaling pathways including key processes involved in cancer development, like proliferation, cell cycle, apoptosis and metastasis formation. Circadian clock in mammals synchronizes cellular and physiological processes by transcriptional-translational feedback loops. Not only miRNAs regulate the levels of key clock genes and clock controlled genes, but also a number of miRNAs exhibit circadian expression. Therefore aberrant circadian rhythms increase risk of colorectal cancer also by altered expression of miRNAs. The main aim of the thesis was to identify miRNAs, which regulate both tumorigenesis and circadian...
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The Role of oncogenic microRNA - 155 and proto - oncogen MYB in chronic lymphocytic leukemia
Vargová, Karina
(EN) Chronic lymphocytic leukemia (B-CLL) represents a disease of mature-like B-cells. Due to failed apoptosis but also due to enhanced proliferative signals, the leukemic B-cells accumulate in the peripheral blood, bone marrow, lymph nodes and spleen. The clinical course of B-CLL is very heterogeneous; in some patients B-CLL progresses very rapidly into an aggressive form. Such patients need therapy sooner while in other patients with indolent B-CLL the onset of therapy takes years. Several standard prognostic and disease progression markers are used for disease staging and monitoring, however a reliable marker that will suggest when to start therapy is unknown. Expression of small, non-coding microRNAs is often deregulated and represent important prognostic markers in variety of cancers including leukemia. Hence in our study we concentrated to miR-155, an important molecule regulating differentiation of hematopoietic cells, inflammation process and antibody production. Its aberrant expression was described in Hodgkin`s as well as in non-Hodgkin`s lymphoma, including indolent lymphoproliferations like B-CLL. Our results confirmed elevated levels of both, primary miR-155 transcript and mature form of miR-155 in our B-CLL patient samples (N=239). The aberrant expression of miR-155 in B-CLL samples...
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