• MeSH
• biologická evoluce * MeSH
• fyziologické jevy MeSH
• genomika MeSH
• lidé MeSH
• molekulární evoluce MeSH
• původ života MeSH
• vědomí MeSH
• život MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Heterogeneous spheroids have recently acquired a prominent position in melanoma research because they incorporate microenvironmental cues relevant for melanoma. In this study, we focused on the analysis of microenvironmental factors introduced in melanoma heterogeneous spheroids by different dermal fibroblasts. We aimed to map the fibroblast diversity resulting from previously acquired damage caused by exposure to extrinsic and intrinsic stimuli. To construct heterogeneous melanoma spheroids, we used normal dermal fibroblasts from the sun-protected skin of a juvenile donor. We compared them to the fibroblasts from the sun-exposed photodamaged skin of an adult donor. Further, we analysed the spheroids by single-cell RNA sequencing. To validate transcriptional data, we also compared the immunohistochemical analysis of heterogeneous spheroids to melanoma biopsies. We have distinguished three functional clusters in primary human fibroblasts from melanoma spheroids. These clusters differed in the expression of (a) extracellular matrix-related genes, (b) pro-inflammatory factors, and (c) TGFβ signalling superfamily. We observed a broader deregulation of gene transcription in previously photodamaged cells. We have confirmed that pro-inflammatory cytokine IL-6 significantly enhances melanoma invasion to the extracellular matrix in our model. This supports the opinion that the aspects of ageing are essential for reliable melanoma 3D modelling in vitro.

• Publikační typ
• časopisecké články MeSH

Hyperactivation of the extracellular signal-regulated kinases (ERK) signaling pathway due to B-Raf mutations is central to melanoma development and progression. Our preliminary data suggest that aberrant ERK pathway activation, although mediated by different mechanisms, is present also in majority of head and neck squamous cell carcinomas (HNSCC). This finding identifies the ERK activation as a key oncogenic event and provides the opportunity to concurrently analyze tumors of different origin but with similar oncogenic alterations. In this proposal, we will determine the role of the pathway in HNSCC and melanoma and compare the modes of its activation in these cancers. We will use biobanked and newly collected tumor samples to perform transcriptome profiling followed by unsupervised clustering analyses. In parallel, we will perform protein expression studies, immunohistochemistry, and kinase activation studies. Obtained tumor-specific characteristics will be related to clinical outcomes including vemurafenib resistance in order to design new prognostic factors for HNSCC and melano...

Hyperaktivace signální dráhy ERK (extracellular signal-regulated kinases) indukovaná mutacemi v genu B-Raf je klíčová pro vývoj a progresi melanomu. Naše předběžné výsledky ukazují, že i v dlaždicových karcinomech hlavy a krku (HNSCC) je dráha ERK aberantně aktivována, i když jinými mechanismy. Aktivace ERK se tak ukazuje jako jeden z klíčových onkogenních procesů v melanomu a HNSCC. To umožňuje současně studovat nádory s různým histogenetickým původem, ale s obdobnými onkogenními změnami. V navrhovaném projektu budeme studovat úlohu ERK dráhy v HNSCC a melanomu a srovnáme mechanismy její aktivace u těchto zhoubných onemocnění. K identifikaci mechanismů aktivace použijeme analýzu genové exprese s následnou shlukovací analýzou a to jak u archivních, tak u nově odebraných biopsií nádorů. Souběžně budeme studovat expresi proteinů, jak kvantitativně tak immunohistochemicky, a aktivaci proteinkináz. Stanovené nádorově-specifické vlastnosti budeme korelovat s klinickými výstupy, včetně rezistence na léčbu vemurafenibem, s cílem navrhnout nové prognostické faktory pro HNSCC a melanom.

• MeSH
• dlaždicobuněčné karcinomy hlavy a krku diagnóza   terapie   MeSH
• lidé MeSH
• MAP kinasový signální systém MeSH
• melanom diagnóza   terapie   MeSH
• nádorové biomarkery MeSH
• prognóza MeSH
• shluková analýza MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé MeSH

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

Aberrant regulation of the cell cycle is a typical feature of all forms of cancer. In head and neck squamous cell carcinoma (HNSCC), it is often associated with the overexpression of cyclin D1 (CCND1). However, it remains unclear how CCND1 expression changes between tumor and normal tissues and whether human papillomavirus (HPV) affects differential CCND1 expression. Here, we evaluated the expression of D-type cyclins in a cohort of 94 HNSCC patients of which 82 were subjected to whole genome expression profiling of primary tumors and paired normal mucosa. Comparative analysis of paired samples showed that CCND1 was upregulated in 18% of HNSCC tumors. Counterintuitively, CCND1 was downregulated in 23% of carcinomas, more frequently in HPV-positive samples. There was no correlation between the change in D-type cyclin expression and patient survival. Intriguingly, among the tumors with downregulated CCND1, one-third showed an increase in cyclin D2 (CCND2) expression. On the other hand, one-third of tumors with upregulated CCND1 showed a decrease in CCND2. Collectively, we have shown that CCND1 was frequently downregulated in HNSCC tumors. Furthermore, regardless of the HPV status, our data suggested that a change in CCND1 expression was alleviated by a compensatory change in CCND2 expression.

• Publikační typ
• časopisecké články MeSH

• MeSH
• CRISPR-Cas systémy genetika   MeSH
• editace genu * etika   MeSH
• etika výzkumu MeSH
• lékařská etika MeSH

• MeSH
• asistovaná reprodukce etika   zákonodárství a právo   MeSH
• chorobopisy normy   zákonodárství a právo   MeSH
• formuláře souhlasu normy   využití   zákonodárství a právo   MeSH
• genetická predispozice k nemoci * genetika   MeSH
• genetické testování * metody   využití   zákonodárství a právo   MeSH
• genom lidský genetika   MeSH
• informovaný souhlas pacienta * etika   normy   zákonodárství a právo   MeSH
• lidé MeSH
• lidská práva zákonodárství a právo   MeSH
• mutační analýza DNA etika   metody   MeSH
• odběr biologického vzorku etika   metody   využití   MeSH
• paternita MeSH
• záznamy jako téma normy   zákonodárství a právo   MeSH
• Check Tag
• lidé MeSH

We announce the completion of the genome sequence of a phenol derivative-degrading bacterium, Rhodococcus erythropolis strain CCM2595. This bacterium is interesting in the context of bioremediation for its capability to degrade phenol, catechol, resorcinol, hydroxybenzoate, hydroquinone, p-chlorophenol, p-nitrophenol, pyrimidines, and sterols.

• Publikační typ
• časopisecké články MeSH

The cluster of pbtTFYRABC genes is carried by plasmid pA81. Its elimination from Achromobacter xylosoxidans A8 resulted in increased sensitivity towards Pb(2+) and Cd(2+). Predicted pbtTRABC products share strong similarities with Pb(2+) uptake transporter PbrT, transcriptional regulator PbrR, metal efflux P1-ATPases PbrA and CadA, undecaprenyl pyrophosphatase PbrB and its signal peptidase PbrC from Cupriavidus metallidurans CH34. Expression of pbtABC or pbtA in a metal-sensitive Escherichia coli GG48 rendered the strain Pb(2+)-, Cd(2+)- and Zn(2+)-tolerant and caused decreased accumulation of the metal ions. Accumulation of Pb(2+), but not of Cd(2+) or Zn(2+), was promoted in E. coli expressing pbtT. Additional genes of the pbt cluster are pbtF and pbtY, which encode the cation diffusion facilitator (CDF)-like transporter and a putative fatty acid hydroxylase of unknown function, respectively. Expression of pbtF did not confer increased metal tolerance upon E. coli GG48, although the protein showed measurable Pb(2+)-efflux activity. Unlike the pbtT promoter, promoters of pbtABC, pbtF and pbtY contain features characteristic of promoters controlled by metal-responsive transcriptional regulators of the MerR family. Upregulation of pbtABC, pbtF and pbtY upon Pb(2+), Cd(2+) and Zn(2+) exposure was confirmed in wild-type Achromobacter xylosoxidans A8. Gel shift assays proved binding of purified PbtR to the respective promoters.

• MeSH
• Achromobacter denitrificans účinky léků   genetika   MeSH
• bakteriální geny MeSH
• Escherichia coli účinky léků   genetika   MeSH
• exprese genu MeSH
• kadmium toxicita   MeSH
• klonování DNA MeSH
• multigenová rodina MeSH
• olovo toxicita   MeSH
• plazmidy MeSH
• regulace genové exprese u bakterií účinky léků   MeSH
• stanovení celkové genové exprese MeSH
• tolerance léku * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

It is generally assumed that human endogenous retroviral elements (HERVs) belong to the class of genomic repetitive nucleotide sequences often called 'junk DNA'. These elements were categorized to families, and members of some of these families (e.g. HERV-H, HERV-W and HERV-K) were shown to be transcribed. These transcriptions were associated with several severe diseases such as mental disorders, AIDS, autoimmune diseases and cancer. In this review we discuss several bioinformatics strategies for genome-wide scan of HERVs transcription using high-throughput RNA sequencing on several platforms. We show that many more HERVs than previously described are transcribed to various levels and we discuss possible implications of these transcriptions.

• MeSH
• endogenní retroviry genetika   MeSH
• genetická transkripce genetika   MeSH
• lidé MeSH
• orgánová specificita MeSH
• sekvenční analýza RNA MeSH
• transpozibilní elementy DNA genetika   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Formation of all metazoan bodies is controlled by a group of selector genes including homeobox genes, highly conserved across the entire animal kingdom. The homeobox genes from Pou and Six classes are key members of the regulation cascades determining development of sensory organs, nervous system, gonads and muscles. Besides using common bilaterian models, more attention has recently been targeted at the identification and characterization of these genes within the basal metazoan phyla. Cnidaria as a diploblastic sister group to bilateria with simple and yet specialized organs are suitable models for studies on the sensory organ origin and the associated role of homeobox genes. In this work, Pou and Six homeobox genes, together with a broad range of other sensory-specific transcription factors, were identified in the transcriptome of hydrozoan jellyfish Craspedacusta sowerbyi. Phylogenetic analyses of Pou and Six proteins revealed cnidarian-specific sequence motifs and contributed to the classification of individual factors. The majority of the Craspedacusta sowerbyi Pou and Six homeobox genes are predominantly expressed in statocysts, manubrium and nerve ring, the tissues with sensory and nervous activities. The described diversity and expression patterns of Pou and Six factors in hydrozoan jellyfish highlight their evolutionarily conserved functions. This study extends the knowledge of the cnidarian genome complexity and shows that the transcriptome of hydrozoan jellyfish is generally rich in homeodomain transcription factors employed in the regulation of sensory and nervous functions.

• MeSH
• faktory domény POU chemie   genetika   metabolismus   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• Hydrozoa genetika   MeSH
• molekulární evoluce MeSH
• orgánová specificita MeSH
• terciární struktura proteinů MeSH
• transkriptom MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH