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First-trimester screening of pregnancy-related complications using plasma exosomal C19MC microRNAs
Špačková, Kamila ; Hromadníková, Ilona (advisor) ; Daňková, Pavlína (referee)
Pregnancy-related complications such as gestational hypertension, preeclampsia, fetal growth restriction, gestational diabetes mellitus, spontaneous preterm birth, and preterm premature rupture of membranes may have severe consequences for both the mother and the child. The development of reliable early screening methods for pregnancy-related complications has therefore been a long-term goal of obstetrics. New possibilities for prenatal diagnostics have opened with the discovery of circulating microRNAs in maternal plasma. MicroRNAs are short, noncoding, 21 to 23 nucleotides long, single-strand RNAs whose main function is to regulate gene expression. During pregnancy, both common and unique miRNAs are expressed by the placenta, amongst them the miRNAs of the C19MC cluster. Several C19MC miRNAs have been shown to display a different expression profile associated with certain pregnancy-related complications. This thesis identifies the plasma exosomal profiles of six C19MC miRNAs (miR-516-5p, miR-517-5p, miR-518b, miR-520a-5p, miR-520h, and miR-525-5p) in patients in their first trimester of gestation who later developed pregnancy-related complications, and compares them with profiles in patients with normal pregnancies.
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Expression and prognostic value of microRNAs in patients with early breast cancer.
Sochor, Marek ; Tesařová, Petra (advisor) ; Slabý, Ondřej (referee) ; Hajdúch, Marián (referee)
Breast cancer is the most frequent cancer in women and is prognostically very heterogeneous. Early breast cancer has an excellent overall prognosis with long-term survival above 90%. In this group we can also find patients with highly unfavourable progress with a risk of future disease relapse. Due to effective anticancer treatment is a main task of precise clinical decision to determine risk of an individual patient in the term of cancer relapse. We can use clinical (tumor diameter, lymph nodes) and pathological markers (grade, ER, PgR, HER2, and Ki-67), all of them have low individual sensitivity and specificity. Molecular tests based on multigene DNA or RNA assays have higher sensitivity and specificity but their interrelated concordance is low. One of the main scientific task is to find almost specific and sensitive prognostic biomarkers. microRNAs are small, highly stable, non-coding RNAs, which regulate tens of mRNAs and proteins inside cells. In cancerogenesis, they could act as oncogenes or tumor supressors as well and affect main steps of initiation and progression of cancer. One of the scientific directions is to determine their prognostic significance. Many experimental and clinical studies defining prognostic significance of miRs in early breast cancer was published but their data were...
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MicroRNA expression in glucocorticoid-treated patients with systemic autoimmune
Uher, Martin ; Kuchařová, Monika (advisor) ; Nováková, Veronika (referee)
Rheumatoid arthritis is the most common joint disease of autoimmune origin. It is accompanied by inflammatory conditions that lead to irreversible changes in the joints, their deformities ending with permanent disability. Treatment of the disease involves routine regimens, surgical, as well as pharmacological treatment, which is necessary for advanced forms. Glucocorticoids play an important role in the therapeutic intervention in the course and progression of the disease. In spite of their anti-inflammatory effect, which is a key to improving the condition of the patient, they have a number of side effects in the long term- use. In this study, we have focused on the impact of these drugs on microRNA expression changes in arthritic patients treated with pulsed doses of glucocorticoids. MicroRNAs are nowadays widely studied due to their possible use as biomarkers in monitoring disease progression and the effect of treatment. MiRNA expression analysis was performed by quantitative real-time PCR array of 754 miRNAs with reverse transcription using stem-loop primers that allow amplification of short sequences that microRNAs are. Data analysis revealed 29 miRNAs differentially expressed at the significance level p ≤ 0.05, 14 miRNAs were at significance level p ≤ 0.025 (respectively 7 miRNAs at p ≤ 0.005...
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MicroRNA expression in glucocorticoid-treated patients with systemic autoimmune
Uher, Martin ; Kuchařová, Monika (advisor) ; Nováková, Veronika (referee)
Rheumatoid arthritis is the most common joint disease of autoimmune origin. It is accompanied by inflammatory conditions that lead to irreversible changes in the joints, their deformities ending with permanent disability. Treatment of the disease involves routine regimens, surgical, as well as pharmacological treatment, which is necessary for advanced forms. Glucocorticoids play an important role in the therapeutic intervention in the course and progression of the disease. In spite of their anti-inflammatory effect, which is a key to improving the condition of the patient, they have a number of side effects in the long term- use. In this study, we have focused on the impact of these drugs on microRNA expression changes in arthritic patients treated with pulsed doses of glucocorticoids. MicroRNAs are nowadays widely studied due to their possible use as biomarkers in monitoring disease progression and the effect of treatment. MiRNA expression analysis was performed by quantitative real-time PCR array of 754 miRNAs with reverse transcription using stem-loop primers that allow amplification of short sequences that microRNAs are. Data analysis revealed 29 miRNAs differentially expressed at the significance level p ≤ 0.05, 14 miRNAs were at significance level p ≤ 0.025 (respectively 7 miRNAs at p ≤ 0.005...
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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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