Myelodysplastic syndromes (MDS) are hematopoietic stem cell disorders with large heterogeneity at the clinical and molecular levels. As diagnostic procedures shift from bone marrow biopsies towards less invasive techniques, circulating small noncoding RNAs (sncRNAs) have become of particular interest as potential novel noninvasive biomarkers of the disease. We aimed to characterize the expression profiles of circulating sncRNAs of MDS patients and to search for specific RNAs applicable as potential biomarkers. We performed small RNA-seq in paired samples of total plasma and plasma-derived extracellular vesicles (EVs) obtained from 42 patients and 17 healthy controls and analyzed the data with respect to the stage of the disease, patient survival, response to azacitidine, mutational status, and RNA editing. Significantly higher amounts of RNA material and a striking imbalance in RNA content between plasma and EVs (more than 400 significantly deregulated sncRNAs) were found in MDS patients compared to healthy controls. Moreover, the RNA content of EV cargo was more homogeneous than that of total plasma, and different RNAs were deregulated in these two types of material. Differential expression analyses identified that many hematopoiesis-related miRNAs (e.g., miR-34a, miR-125a, and miR-150) were significantly increased in MDS and that miRNAs clustered on 14q32 were specifically increased in early MDS. Only low numbers of circulating sncRNAs were significantly associated with somatic mutations in the SF3B1 or DNMT3A genes. Survival analysis defined a signature of four sncRNAs (miR-1237-3p, U33, hsa_piR_019420, and miR-548av-5p measured in EVs) as the most significantly associated with overall survival (HR = 5.866, p < 0.001). In total plasma, we identified five circulating miRNAs (miR-423-5p, miR-126-3p, miR-151a-3p, miR-125a-5p, and miR-199a-3p) whose combined expression levels could predict the response to azacitidine treatment. In conclusion, our data demonstrate that circulating sncRNAs show specific patterns in MDS and that their expression changes during disease progression, providing a rationale for the potential clinical usefulness of circulating sncRNAs in MDS prognosis. However, monitoring sncRNA levels in total plasma or in the EV fraction does not reflect one another, instead, they seem to represent distinctive snapshots of the disease and the data should be interpreted circumspectly with respect to the type of material analyzed.

• MeSH
• azacytidin farmakologie   MeSH
• biologické markery krev   MeSH
• biologické modely MeSH
• editace RNA genetika   MeSH
• extracelulární vezikuly metabolismus   MeSH
• Kaplanův-Meierův odhad MeSH
• lidé MeSH
• malá nekódující RNA krev   genetika   MeSH
• mikro RNA genetika   metabolismus   MeSH
• multivariační analýza MeSH
• mutace genetika   MeSH
• myelodysplastické syndromy krev   genetika   patologie   MeSH
• prognóza MeSH
• proporcionální rizikové modely MeSH
• regulace genové exprese MeSH
• reprodukovatelnost výsledků MeSH
• signální transdukce genetika   MeSH
• výsledek terapie MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The circulating human transcriptome, which includes both coding and non-coding RNA (ncRNA) molecules, represents a rich source of potential biomarkers for colorectal cancer (CRC) that has only recently been explored. In particular, the release of RNA-containing extracellular vesicles (EVs), in a multitude of different in vitro cell systems and in a variety of body fluids, has attracted wide interest. The role of RNA species in EVs is still not fully understood, but their capacity to act as a form of distant communication between cells and their higher abundance in association with cancer demonstrated their relevance. In this review, we report the evidence from both in vitro and human studies on microRNAs (miRNAs) and other ncRNA profiles analysed in EVs in relation to CRC as diagnostic, prognostic and predictive markers. The studies so far highlighted that, in exosomes, the most studied category of EVs, several miRNAs are able to accurately discriminate CRC cases from controls as well as to describe the progression of the disease and its prognosis. Most of the time, the in vitro findings support the miRNA profiles detected in human exosomes. The expression profiles measured in exosomes and other EVs differ and, interestingly, there is a variability of expression also among different subsets of exosomes according to their proteic profile. On the other hand, evidence is still limited for what concerns exosome miRNAs as early diagnostic and predictive markers of treatment. Several other ncRNAs that are carried by exosomes, mostly long ncRNAs and circular RNAs, seem also to be dysregulated in CRC. Besides various technical challenges, such as the standardisation of EVs isolation methods and the optimisation of methodologies to characterise the whole spectrum of RNA molecules in exosomes, further studies are needed in order to elucidate their relevance as CRC markers.

• MeSH
• exozómy genetika   metabolismus   MeSH
• extracelulární vezikuly genetika   metabolismus   MeSH
• kolorektální nádory krev   diagnóza   genetika   metabolismus   MeSH
• lidé MeSH
• mikro RNA genetika   metabolismus   MeSH
• nádorové biomarkery genetika   metabolismus   MeSH
• nekódující RNA genetika   metabolismus   MeSH
• progrese nemoci MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Early detection of colorectal cancer (CRC) is the key for prevention and the ability to impact long-term survival of CRC patients. Current CRC screening modalities are inadequate for global application because of low sensitivity and specificity in case of conventional stool-based screening tests, and high costs and a low participation compliance in colonoscopy. An accurate stool- or blood-based screening test with use of innovative biomarkers is an appealing alternative as it is non-invasive and poses minimal risk to patients. It is easy to perform, can be repeated at shorter intervals, and therefore would likely lead to a much higher compliance rates. Non-coding RNAs (ncRNAs) have recently gained attention because of their involvement in different biological processes, such as proliferation, differentiation, migration, angiogenesis and apoptosis. An increasing number of studies have demonstrated that mutations or abnormal expression of ncRNAs are closely associated with various cancers, including CRC. The discovery that ncRNAs (mainly microRNAs) are stable in stool and in blood plasma and serum presents the opportunity to develop novel strategies taking advantage of circulating ncRNAs as early diagnostic biomarkers of CRC. This chapter is a comprehensive examination of aberrant ncRNAs expression levels in tumor tissue, stool and blood of CRC patients and a summary of the current findings on ncRNAs, including microRNAs, small nucleolar RNAs, small nuclear RNAs, Piwi-interacting RNAs, circular RNAs and long ncRNAs in regards to their potential usage for screening or early detection of CRC.

• MeSH
• adenokarcinom chemie   diagnóza   genetika   MeSH
• adenom chemie   diagnóza   genetika   MeSH
• časná detekce nádoru metody   MeSH
• feces chemie   MeSH
• kolorektální nádory chemie   diagnóza   genetika   MeSH
• krevní plazma MeSH
• lidé MeSH
• nádorové biomarkery analýza   krev   MeSH
• nekódující RNA analýza   krev   MeSH
• pacientův souhlas se zdravotní péčí MeSH
• polypy tlustého střeva chemie   diagnóza   genetika   MeSH
• regulace genové exprese u nádorů MeSH
• senzitivita a specificita MeSH
• sérum MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Identifying patients likely to develop breast cancer recurrence remains a challenge. Thus, the discovery of biomarkers capable of diagnosing recurrence is of the utmost importance. MiRNAs are small, non-coding RNA molecules which are known to regulate genetic expression and have previously demonstrated relevance as biomarkers in malignancy. To perform a systematic review evaluating the role of miRNAs in predicting breast cancer recurrence. A formal systematic search of PubMed, Scopus, Web of Science, and Cochrane databases was performed. This search was performed according to the Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) checklist. A total of 19 studies involving 2287 patients were included. These studies identified 44 miRNAs which predicted breast cancer recurrence. Results from nine studies assessed miRNAs in tumour tissues (47.4%), eight studies included circulating miRNAs (42.1%), and two studies assessed both tumour and circulating miRNAs (10.5%). Increased expression of 25 miRNAs were identified in patients who developed recurrence, and decreased expression of 14 miRNAs. Interestingly, five miRNAs (miR-17-5p, miR-93-5p, miR-130a-3p, miR-155, and miR-375) had discordant expression levels, with previous studies indicating both increased and reduced expression levels of these biomarkers predicting recurrence. MiRNA expression patterns have the ability to predict breast cancer recurrence. These findings may be used in future translational research studies to identify patients with breast cancer recurrence to improve oncological and survival outcomes for our prospective patients.

• MeSH
• cirkulující mikroRNA * MeSH
• lidé MeSH
• malá nekódující RNA * MeSH
• mikro RNA * genetika   metabolismus   MeSH
• nádorové biomarkery genetika   MeSH
• nádory prsu * diagnóza   genetika   metabolismus   MeSH
• prospektivní studie MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• systematický přehled MeSH

miRNAs are short noncoding RNAs that post-transcriptionally regulate gene expression. miRNAs' ability to inhibit translation of oncogenes and tumor suppressor genes implies they have an involvement in carcinogenesis. Specific miRNA expression signatures have been identified in a variety of human cancers. More recently, occurrence of miRNAs in the blood serum and plasma of humans has been repeatedly observed. miRNA levels in serum and plasma are more stable, reproducible and consistent among individuals of the same species in comparison with other circulating nucleic acids. Circulating miRNAs have been successfully evaluated in a wide range of solid cancers as promising novel noninvasive biomarkers of early disease onset or relapse. Here we describe the origin of circulating miRNAs, principles of their immense stability and proposed functions, and comprehensively summarize studies focusing on their significance in the most frequently studied cancer types in this regard, including breast, colorectal, lung and prostate cancer.

• MeSH
• kolorektální nádory krev   MeSH
• lidé MeSH
• mikro RNA krev   MeSH
• nádorové biomarkery krev   MeSH
• nádory plic krev   MeSH
• nádory prostaty krev   MeSH
• nádory prsu krev   MeSH
• stabilita RNA MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Na rozdíl od klasických biopsií představují tekuté biopsie jemnější, více dostupný, méně bolestivý a komplexnější přístup, který je možné opakovat častěji, a umožňují tak získání biologicky relevantních informací o celém nádoru, ale i monitorování léčebné odpovědi a detekci minimální reziduální choroby. To je možné díky tomu, že periferní krev obsahuje nejen cirkulující nádorové buňky, ale také různé cirkulující molekuly nukleových kyselin (mikroRNA, mimobuněčné DNA, dlouhé nekódující RNA atd.). Mnohočetný myelom je geneticky heterogenní onemocnění charakterizované multifokálními nádorovými ložisky v kostní dřeni, ale i fokálními ložisky mimo kostní dřeň. Biopsie kostní dřeně z jednoho místa ovlivňuje molekulární profil, který je limitovaný místem odběru, protože taková biopsie neposkytne informaci ze všech klonů. Navíc během progrese nemoci a léčby se molekulární profil mění a subklony buněk mnohočetného myelomu se mohou stát rezistentními k léčbě. Navíc, různé klony, které se objevují v extramedulárních oblastech, které se navíc nenachází v kostní dřeni, reagují na léčbu jinak a přímo ovlivňují přežití pacientů. Pro nemoci jako je mnohočetný myelom se vyšetření pomocí tekuté biopsie jeví jako relevantní a nutný další krok.

Unlike bone marrow biopsies, liquid biopsies represent a gentler, more accessible, less painful, repeatable and more comprehensive approach to get biologically relevant information about the entire tumor but also about treatment response and level of minimal residual disease. This is all possible since peripheral blood contains not only circulating tumor cells but also many circulating molecules of nucleic acids (microRNA, cell-free DNA, long non-coding RNA etc.). Multiple myeloma is a genetically heterogeneous disease characterized by multifocal tumor deposits in the bone marrow but also focal lesions elsewhere. Single-site biopsy of the bone marrow creates a sampling bias that provides a limited molecular profile as the biopsy cannot capture all subclones. Moreover, during disease progression and treatment, molecular profile is changed and subclones of multiple myeloma cells resistant to treatment are formed. Likewise, various clones found in extramedullary sites that are not present in the bone marrow respond differently to treatment directly influencing survival of patients. Thus, liquid biopsies seem to be a relevant and necessary next step for diseases such as multiple myeloma.

• MeSH
• biopsie * metody   využití   MeSH
• DNA analýza   krev   MeSH
• lidé MeSH
• mikro RNA analýza   fyziologie   MeSH
• mnohočetný myelom * mikrobiologie   patologie   MeSH
• nekódující RNA fyziologie   MeSH
• odběr vzorku krve MeSH
• prognóza MeSH
• reziduální nádor diagnóza   patologie   MeSH
• RNA dlouhá nekódující fyziologie   MeSH
• tekutá biopsie MeSH
• vyšetřování kostní dřeně MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

The endothelial-mesenchymal transition (EndMT) of endothelial progenitor cells (EPCs) plays a notable role in pathological vascular remodeling. Emerging evidence indicated that long non-coding RNA-regulator of reprogramming (linc-ROR) can promote epithelial-mesenchymal transition (EMT) in a variety of cancer cells. Nevertheless, the function of linc-ROR in EPC EndMT has not been well elucidated. The present study investigated the effect and possible mechanisms of function of linc-ROR on the EndMT of EPCs. A linc-ROR overexpression lentiviral vector (LV linc-ROR) or a linc-ROR short hairpin RNA lentiviral vector (LV-shlinc-ROR) was used to up or downregulate linc-ROR expression in EPCs isolated from human umbilical cord blood. Functional experiments demonstrated that LV-linc-ROR promoted the proliferation and migration of EPCs, but inhibited EPC angiogenesis in vitro. In the meantime, reverse transcription-quantitative PCR and western blotting results showed that the expression of the endothelial cell markers vascular endothelial-cadherin and CD31 was decreased, while the expression of the mesenchymal cell markers ?-smooth muscle actin and SM22? was increased at both mRNA and protein levels in LV-linc-ROR-treated EPCs, indicating that linc-ROR induced EPC EndMT. Mechanistically, the dual-luciferase reporter assay demonstrated that microRNA (miR/miRNA)-145 was a direct target of linc-ROR, and miR-145 binds to the 3'-untranslated region of Smad3. Moreover, LV-shlinc-ROR increased the expression of miR-145, but decreased the expression of Smad3. In conclusion, linc-ROR promotes EPC EndMT, which may be associated with the miR-145/Smad3 signaling pathway. Keywords: Endothelial progenitor cells, Endothelial to mesenchymal transition, Linc-ROR, MiR-145, Atherosclerosis.

• MeSH
• endotel-mezenchymální transformace MeSH
• endoteliální progenitorové buňky * metabolismus   MeSH
• epitelo-mezenchymální tranzice * MeSH
• kultivované buňky MeSH
• lidé MeSH
• mikro RNA * metabolismus   genetika   MeSH
• pohyb buněk fyziologie   MeSH
• proliferace buněk MeSH
• protein Smad3 * metabolismus   genetika   MeSH
• RNA dlouhá nekódující * genetika   metabolismus   MeSH
• signální transdukce * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Plasmocelulární leukemie (PCL) je nejagresivnější formou monoklonálních gamapatií. Je to raritní onemocnění, s incidencí v Evropě jen zhruba 0,04 případů na 100 000 obyvatel za rok. Diagnostická kritéria PCL jsou založena na více než 20 % cirkulujících plazmatických buňkách v periferní krvi a jejich absolutním počtem vyšším než 2x109/l. Tyto hodnoty byly však nastaveny v roce 1974 a kvůli malému množství pacientů nebyly nikdy řádně validovány a mohou podhodnocovat opravdový počet pacientů s PCL. Je tedy možné, že nižší hladina plasmocytózy je dostatečná pro definici této choroby. Zároveň, nekódující molekuly RNA přímo regulují expresi genů, které mohou ovlivnit migraci plazmatických buněk z kostní dřeně do periferní krve. Tento grant se zaměří na redefinování diagnostických kritérií PCL na základě flowcytometrické analýzy cirkulujících plazmatických buněk, změn jejich hladin v závislosti na léčbě a na analýzu nekódujících molekul RNA.; Plasma cell leukemia (PCL) is the most aggressive form of monoclonal gammopathies. It is a rare disease with incidence in Europe of only 0.04 cases per 100 000 people per year. The PCL diagnostic criteria are based on more than 20% of plasma cells in peripheral blood and their absolute count greater than 2x109/L. However, these values were set in 1974; due to very small number of patients, they were not properly validated and may underestimate the real number of PCL patients. Thus, it is possible that lower degree of peripheral plasmocytosis is sufficient to define the disease. At the same time, the so-called non-coding RNA molecules directly regulate expression of genes; thus, the effect of these molecules on changes that allow cPC to migrate out of the bone marrow will be analyzed. This grant aims to redefine diagnostics criteria of PCL based on flowcytometric analysis of circulating plasma cells, changes in their amounts in connection to treatment and on analysis of non-coding RNA molecules.

• MeSH
• exprese genu MeSH
• nekódující RNA MeSH
• plazmocelulární leukemie diagnóza   MeSH
• průtoková cytometrie MeSH
• tekutá biopsie MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• onkologie
• hematologie a transfuzní lékařství
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

MicroRNAs (miRNAs) comprise a large group of small noncoding RNAs within a heterogeneous entity of noncoding RNAs, forming potent functional tools regulating the crucial biological processes within cells and the body. Cell-free miRNAs have become one of the novel promising diagnostic, predictive, and prognostic biomarkers for various diseases extensively investigated in recent years. This is due to their presence within extracellular fractions of various body fluids suggesting their potential as noninvasive "liquid biopsy" in case of their dysregulated expression.Among the body fluids, blood plasma and serum along with urine are the most commonly investigated sources of various types of cell-free miRNAs. Another body fluid, i.e., ascites (effusion, peritoneal/pleural fluid) may be the clinically important fluid particularly associated with carcinogenesis in ovarian carcinomas and hepatocellular carcinomas or in case of liver cirrhosis.Here, we provide a protocol for an expression profiling study based on qPCR analyses aimed at finding novel candidate miRNAs via small-scale or large-scale screening and evaluation experiments using liquid biopsies of blood plasma, ascites, and urine. Using this approach may be worth in cases where no (or limited) information is available on miRNA expression in particular diseases and geographic regions, for validation of previously published miRNAs with promising diagnostic potential, particularly in situations where follow-up study is aimed at validating miRNAs coming from (micro) array or NGS experiments, or where funding for large-scale experiments is not available. We demonstrate that assessment of plasma, ascites, and urine miRNAs expression may represent a feasible method to explore the potential for finding novel diagnostic, predictive, and prognostic biomarkers for various diseases.

• MeSH
• ascites MeSH
• biologické markery MeSH
• cirkulující mikroRNA * MeSH
• krevní plazma chemie   MeSH
• lidé MeSH
• mikro RNA * genetika   MeSH
• nádorové biomarkery MeSH
• následné studie MeSH
• tělesné tekutiny * metabolismus   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Východiska: Jedním z moderních přístupů identifikace biomarkerů nádorových onemocnění, a to jak na tkáňové úrovni, tak i v tělních tekutinách, je profilování exprese mikroRNA (miRNA). miRNA tvoří skupinu téměř 3 000 krátkých, 18-25 nukleotidů dlouhých nekódujících RNA. Slouží jako regulační prvky, které řídí expresi genů na posttranskripční úrovni, tj. na úrovni molekul mRNA. Schopnost miRNA inhibovat translaci či indukovat degradaci onkogenů a nádorových supresorů je podstatou jejich zapojení do procesů kancerogeneze. Důkazů o funkcích miRNA v regulaci procesů, jako jsou apoptóza, buněčná proliferace, diferenciace či invazivita, neustále přibývá. Analýza expresních profilů miRNA je proto stále častěji využívána pro účely molekulární diagnostiky nádorových onemocnění, analogicky jako je tomu u studií založených na profilování kódujících RNA. Z hlediska analytického využití je podstatná skutečnost, že miRNA jsou vysoce stabilní v tělních tekutinách vč. slin a vyskytují se zde v relativně vysokých hladinách. miRNA ve slinách již byly pro diagnostické účely úspěšně testovány u řady nádorových onemocnění, přičemž hlavní výhodou slin jako biologického materiálu je skutečnost, že jsou získatelné zcela neinvazivně. Cíl: Cílem přehledového článku je shrnout dosavadní míru poznání z oblasti cirkulujících miRNA u nádorových onemocnění se zaměřením na využití miRNA ve slinách pro účely onkologické diagnostiky.

Background: A modern approach to identify biomarkers of solid cancers in tissues and body fluids is based on microRNA (miRNA) expression profiling. miRNAs are a group of approximately 3.000 short noncoding RNAs containing 18-25 nucleotides that regulate gene expression at the post-transcriptional (mRNA) level. The abilities of miRNAs to inhibit the translation or induce degradation of oncogenes and tumor suppressors indicate that they are involved in carcinogenesis. There is increasing evidence that miRNAs regulate apoptosis, cell proliferation, differentiation, and invasion. miRNA expression profiles are therefore often analyzed for molecular diagnostics of solid cancers, similar to analyses based on mRNA profiling. It is important that miRNAs are highly stable and present at high levels in body fluids, including saliva, for analytic usage. miRNAs in saliva have been successfully tested as potential diagnostic biomarkers of many solid cancers. The main advantage of these miRNAs is that saliva samples can be collected non-invasively. Aim: This review aims to summarize current knowledge of circulating miRNAs in solid cancers, with a focus on the use of miRNAs in saliva for oncology diagnostics.

• MeSH
• cirkulující mikroRNA MeSH
• hormony MeSH
• lidé MeSH
• mikro RNA * analýza   klasifikace   MeSH
• nádorové biomarkery MeSH
• nádory hlavy a krku diagnóza   MeSH
• nádory jícnu diagnóza   MeSH
• nádory slinivky břišní diagnóza   MeSH
• nádory * diagnóza   MeSH
• posttranskripční úpravy RNA MeSH
• regulace genové exprese MeSH
• sliny * chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH