Doporučené postupy klinické péče o nosiče patogenních variant v klinicky relevantních genech predisponujících ke vzniku Lynchova syndromu a karcinomu kolorekta definují kroky primární a sekundární prevence, která by měla být osobám ve vysokém riziku vzniku dědičných nádorů v ČR poskytnuta. Tvorba doporučených postupů byla organizována pracovní skupinou onkogenetiky Společnosti lékařské genetiky a genomiky při České lékařské společnosti J. E. Purkyně ve spolupráci se zástupci onkologie, onkogynekologie a gastroenterologie. Doporučené postupy vycházejí z aktuálních doporučení National Comprehensive Cancer Network (NCCN), Evropské společnosti pro klinickou onkologii (ESMO) a zohledňují kapacitní možnosti našeho zdravotnictví.

The guidelines for clinical practice for carriers of pathogenic variants in clinically relevant genes predisposing to Lynch syndrome and colorectal cancer define the steps of primary and secondary prevention that should be provided to the individuals at high risk of developing hereditary cancer in the Czech Republic. The drafting of the guidelines was organized by the Oncogenetics Working Group of the Society for Medical Genetics and Genomics of J. E. Purkyně Czech Medical Society, in cooperation with representatives of oncology, oncogynecology, and gastroenterology. The guidelines are based on the current recommendations of the National Comprehensive Cancer Network (NCCN), European Society of Medical Oncology (ESMO) and take into account the capacity of the Czech healthcare system.

• MeSH
• adhezní molekula epiteliálních buněk genetika   MeSH
• dědičné nepolypózní kolorektální nádory genetika   MeSH
• genetická predispozice k nemoci * genetika   MeSH
• kolorektální nádory * genetika   MeSH
• mismatch repair endonukleáza PMS2 genetika   MeSH
• MutL homolog 1 genetika   MeSH
• směrnice pro lékařskou praxi jako téma MeSH
• zárodečné mutace genetika   MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH
• Geografické názvy
• Česká republika MeSH

Cauda equina neuroendocrine tumors (CENETs) are neoplasms of uncertain histogenesis with overlapping features between those of paragangliomas (PGs) and visceral neuroendocrine tumors (NETs). We have explored their biological relationship to both subsets of neuroendocrine neoplasms. The clinical and radiological features of a cohort of 23 CENETs were analyzed. A total of 21 cases were included in tissue microarrays, along with a control group of 38 PGs and 83 NETs. An extensive panel of antibodies was used to assess epithelial phenotype (cytokeratins, E-cadherin, EpCAM, Claudin-4, EMA, CD138), neuronal and neuroendocrine features (synaptophysin, chromogranin A, INSM1, neurofilaments, NeuN, internexin-α, calretinin), chromaffin differentiation (GATA3, Phox2b, tyrosine hydroxylase), and possible histogenesis (Sox2, T-brachyury, Oct3/4, Sox10). The cohort included 5 women (22%) and 18 men (78%). The average age at the time of surgery was 48.3 years (range from 21 to 80 years). The average diameter of the tumors was 39.27 mm, and invasion of surrounding structures was observed in 6/21 (29%) tumors. Follow-up was available in 16 patients (median 46.5 months). One tumor recurred after 19 months. No metastatic behavior and no endocrine activity were observed. Compared to control groups, CENETs lacked expression of epithelial adhesion molecules (EpCAM, CD138, E-cadherin, Claudin-4), and at the same time, they lacked features of chromaffin differentiation (GATA3, Phox2b, tyrosine hydroxylase). We observed no loss of SDHB. Cytokeratin expression was present in all CENETs. All the CENETs showed variable cytoplasmic expression of T-brachyury and limited nuclear expression of Sox2. These findings support the unique nature of the neoplasm with respect to NETs and PGs.

• MeSH
• adhezní molekula epiteliálních buněk MeSH
• cauda equina * metabolismus   patologie   chirurgie   MeSH
• claudin-4 MeSH
• lidé MeSH
• lokální recidiva nádoru patologie   MeSH
• nádory centrálního nervového systému * patologie   MeSH
• neuroendokrinní nádory * patologie   MeSH
• paragangliom * MeSH
• represorové proteiny MeSH
• transkripční faktory MeSH
• tyrosin-3-monooxygenasa MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Circulating tumor cells (CTCs) represent one of the promising diagnostic targets in monitoring of lung adenocarcinoma progression and malignant transformation. Here we propose a microfluidic chip (MLC Chip) which will enable to isolate (count) population of CTCs from peripheral blood sample of histologically proved adenocarcinoma stage IIb-IV patients and to sort them on their respective phenotype (epithelial/mesenchymal) on the level of the individual cells with the possibility of subsequent cell release for cultivation and/or gene expression profiling. The MLC Chip with filtration and single-cell sorting unit will be presented along with the novel types of binding proteins for EpCAM and N-Cadherin markers for CTCs on-chip capture. The overall goal is to evaluate the clinical relevance of different CTC phenotypes on early diagnostics of lung adenocarcinoma, correlate their occurrence with the carcinoma stage and progression, divide a population of patients in high/low risk groups, predict diagnosis and finally to adjust a medical treatment for individual patients.

Cirkulující nádorové buňky (CTC) jsou jedním z potenciálně významných diagnostických markerů pro monitorování metastatické progrese adenokarcinomu plic. Projekt je zaměřen na vývoj mikrofluidního čipu (MLC čip), který umožní izolovat (kvantifikovat) populace CTC buněk ve vzorku periferní krve pacientů s histologicky prokázaným adenokarcinomem plic fáze IIb-IV a rozdělit je dle jejich fenotypového projevu (epiteliální/mesenchymální) na úrovni jednotlivých buněk, s možností jejich následného uvolnění z čipu pro in vitro kultivaci, popř. analýzu genové exprese. MLC čip se bude skládat z filtrační a buněčné sortovací jednotky s imobilizovanými vazebnými proteiny pro zachycení buněk pomocí povrchových membránových markerů EpCAM a N-cadherinu. Hlavním cílem projektu je zhodnocení klinické relevance různých fenotypů CTC buněk na včasnou diagnostiku adenokarcinomu plic, korelovat je se stádiem onemocnění, rozdělit pacienty na high/low risk skupiny, predikovat diagnózu a ve výsledku uzpůsobit léčbu z hlediska individuálních potřeb pacienta.

• MeSH
• adenokarcinom plic diagnóza   MeSH
• adhezní molekula epiteliálních buněk MeSH
• fenotyp MeSH
• kadheriny MeSH
• laboratoř na čipu MeSH
• lidé MeSH
• mikročipová analýza MeSH
• nádorové biomarkery MeSH
• nádorové cirkulující buňky MeSH
• ribozomy MeSH
• Check Tag
• lidé MeSH

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

• MeSH
• adhezní molekula epiteliálních buněk metabolismus   MeSH
• dospělí MeSH
• galaktosyltransferasy genetika   MeSH
• kolorektální nádory metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• sfingolipidy metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The autoimmune regulator (Aire) serves an essential function for T cell tolerance by promoting the "promiscuous" expression of tissue antigens in thymic epithelial cells. Aire is also detected in rare cells in peripheral lymphoid organs, but the identity of these cells is poorly understood. Here, we report that Aire protein-expressing cells in lymph nodes exhibit typical group 3 innate lymphoid cell (ILC3) characteristics such as lymphoid morphology, absence of "classical" hematopoietic lineage markers, and dependence on RORγt. Aire+ cells are more frequent among lineage-negative RORγt+ cells of peripheral lymph nodes as compared with mucosa-draining lymph nodes, display a unique Aire-dependent transcriptional signature, express high surface levels of MHCII and costimulatory molecules, and efficiently present an endogenously expressed model antigen to CD4+ T cells. These findings define a novel type of ILC3-like cells with potent APC features, suggesting that these cells serve a function in the control of T cell responses.

• MeSH
• adhezní molekula epiteliálních buněk metabolismus   MeSH
• antigen prezentující buňky imunologie   MeSH
• antigeny CD11 metabolismus   MeSH
• fenotyp MeSH
• genetická transkripce MeSH
• histokompatibilita - antigeny třídy II metabolismus   MeSH
• jaderné receptory - podrodina 1, skupina F, člen 3 metabolismus   MeSH
• lymfatické uzliny cytologie   MeSH
• lymfocyty imunologie   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši knockoutované MeSH
• myši MeSH
• přirozená imunita MeSH
• regulace genové exprese MeSH
• transkripční faktory genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: During heart development, it has been hypothesized that apoptosis of atrioventricular canal myocardium and replacement by fibrous tissue derived from the epicardium are imperative to develop a mature atrioventricular conduction. To test this, apoptosis was blocked using an established caspase inhibitor and epicardial growth was delayed using the experimental epicardial inhibition model, both in chick embryonic hearts. RESULTS: Chicken embryonic hearts were either treated with the peptide caspase inhibitor zVAD-fmk by intrapericardial injection in ovo (ED4) or underwent epicardial inhibition (ED2.5). Spontaneously beating embryonic hearts isolated (ED7-ED8) were then stained with voltage-sensitive dye Di-4-ANEPPS and imaged at 0.5-1 kHz. Apoptotic cells were quantified (ED5-ED7) by whole-mount LysoTracker Red and anti-active caspase 3 staining. zVAD-treated hearts showed a significantly increased proportion of immature (base to apex) activation patterns at ED8, including ventricular activation originating from the right atrioventricular junction, a pattern never observed in control hearts. zVAD-treated hearts showed decreased numbers of apoptotic cells in the atrioventricular canal myocardium at ED7. Hearts with delayed epicardial outgrowth showed also increased immature activation patterns at ED7.5 and ED8.5. However, the ventricular activation always originated from the left atrioventricular junction. Histological examination showed no changes in apoptosis rates, but a diminished presence of atrioventricular sulcus tissue compared with controls. CONCLUSIONS: Apoptosis in the atrioventricular canal myocardium and controlled replacement of this myocardium by epicardially derived HCN4-/Trop1- sulcus tissue are essential determinants of mature ventricular activation pattern. Disruption can lead to persistence of accessory atrioventricular connections, forming a morphological substrate for ventricular pre-excitation. Developmental Dynamics 247:1033-1042, 2018. © 2018 Wiley Periodicals, Inc.

• MeSH
• adhezní molekula epiteliálních buněk MeSH
• apoptóza * MeSH
• hyperpolarizační iontové kanály řízené cyklickými nukleotidy MeSH
• kuřecí embryo MeSH
• perikard * MeSH
• preexcitační syndromy etiologie   MeSH
• převodní systém srdeční patofyziologie   MeSH
• remodelace komor * MeSH
• remodelace síní * MeSH
• zvířata MeSH
• Check Tag
• kuřecí embryo MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The identification of fibroblasts and cancer-associated fibroblasts from human cancer tissue using surface markers is difficult, especially because the markers used currently are usually not expressed solely by fibroblasts, and the identification of fibroblast-specific surface molecules is still under investigation. It was aimed to compare three commercially available antibodies in the detection of different surface epitopes of fibroblasts (anti-fibroblast, fibroblast activation protein α, and fibroblast surface protein). The specificity of their expression, employing fibroblast cell lines and tumor-derived fibroblasts from breast and prostate tissues was investigated. Both the established fibroblast cell line HFF-1 and ex vivo primary fibroblasts isolated from breast and prostate cancer tissues expressed the tested surface markers to different degrees. Surprisingly, those markers were expressed also by permanent cell lines of epithelial origin, both benign and cancer-derived (breast-cell lines MCF 10A, HMLE and prostate-cell lines BPH-1, DU 145, and PC-3). The expression of fibroblast activation protein α increased on the surface of previously described models of epithelial cells undergoing epithelial-to-mesenchymal transition in response to treatment with TGF-β1. To prove the co-expression of the fibroblast markers on cells of epithelial origin, we used freshly dissociated human prostate and breast cancer tissues. The results confirmed the co-expression of anti-fibroblast and fibroblast surface protein on CD31/CD45-negative/EpCAM-positive epithelial cells. In summary, our data support the findings that the tested fibroblast markers are not fibroblast specific and may be expressed also by cells of epithelial origin (e.g., cells undergoing EMT). Therefore, the expression of these markers should be interpreted with caution, and the combination of several epitopes for both positive (anti-fibroblast or fibroblast activation protein α) and negative (EpCAM) identification of fibroblasts from breast and prostate tumor tissues is advised. © 2017 International Society for Advancement of Cytometry.

• MeSH
• adhezní molekula epiteliálních buněk metabolismus   MeSH
• antigeny CD31 metabolismus   MeSH
• antigeny CD45 metabolismus   MeSH
• biologické markery metabolismus   MeSH
• buňky PC-3 MeSH
• epitelo-mezenchymální tranzice fyziologie   MeSH
• epitelové buňky metabolismus   MeSH
• fibroblasty metabolismus   MeSH
• lidé MeSH
• membránové proteiny metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory prostaty metabolismus   MeSH
• nádory prsu metabolismus   MeSH
• serinové endopeptidasy metabolismus   MeSH
• transformující růstový faktor beta1 metabolismus   MeSH
• želatinasy metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

AGR2 is an oncogenic endoplasmic reticulum (ER)-resident protein disulfide isomerase. AGR2 protein has a relatively unique property for a chaperone in that it can bind sequence-specifically to a specific peptide motif (TTIYY). A synthetic TTIYY-containing peptide column was used to affinity-purify AGR2 from crude lysates highlighting peptide selectivity in complex mixtures. Hydrogen-deuterium exchange mass spectrometry localized the dominant region in AGR2 that interacts with the TTIYY peptide to within a structural loop from amino acids 131-135 (VDPSL). A peptide binding site consensus of Tx[IL][YF][YF] was developed for AGR2 by measuring its activity against a mutant peptide library. Screening the human proteome for proteins harboring this motif revealed an enrichment in transmembrane proteins and we focused on validating EpCAM as a potential AGR2-interacting protein. AGR2 and EpCAM proteins formed a dose-dependent protein-protein interaction in vitro Proximity ligation assays demonstrated that endogenous AGR2 and EpCAM protein associate in cells. Introducing a single alanine mutation in EpCAM at Tyr251 attenuated its binding to AGR2 in vitro and in cells. Hydrogen-deuterium exchange mass spectrometry was used to identify a stable binding site for AGR2 on EpCAM, adjacent to the TLIYY motif and surrounding EpCAM's detergent binding site. These data define a dominant site on AGR2 that mediates its specific peptide-binding function. EpCAM forms a model client protein for AGR2 to study how an ER-resident chaperone can dock specifically to a peptide motif and regulate the trafficking a protein destined for the secretory pathway.

• MeSH
• adhezní molekula epiteliálních buněk genetika   metabolismus   MeSH
• lidé MeSH
• MFC-7 buňky MeSH
• peptidy metabolismus   MeSH
• proteiny genetika   metabolismus   MeSH
• protoonkogenní proteiny c-mdm2 metabolismus   MeSH
• rekombinantní proteiny metabolismus   MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies. Its dismal prognosis is often attributed to the presence of cancer stem cells (CSCs) that have been identified in PDAC using various markers. However, the co-expression of all of these markers has not yet been evaluated. Furthermore, studies that compare the expression levels of CSC markers in PDAC tumor samples and in cell lines derived directly from those tumors are lacking. Here, we analyzed the expression of putative CSC markers-CD24, CD44, epithelial cell adhesion molecule (EpCAM), CD133, and nestin-by immunofluorescence, flow cytometry and quantitative PCR in 3 PDAC-derived cell lines and by immunohistochemistry in 3 corresponding tumor samples. We showed high expression of the examined CSC markers among all of the cell lines and tumor samples, with the exception of CD24 and CD44, which were enriched under in vitro conditions compared with tumor tissues. The proportions of cells positive for the remaining markers were comparable to those detected in the corresponding tumors. Co-expression analysis using flow cytometry revealed that CD24+/CD44+/EpCAM+/CD133+ cells represented a significant population of the cells (range, 43 to 72%) among the cell lines. The highest proportion of CD24+/CD44+/EpCAM+/CD133+ cells was detected in the cell line derived from the tumor of a patient with the shortest survival. Using gene expression profiling, we further identified the specific pro-tumorigenic expression profile of this cell line compared with the profiles of the other two cell lines. Together, CD24+/CD44+/EpCAM+/CD133+ cells are present in PDAC cell lines derived from primary tumors, and their increased proportion corresponds with a pro-tumorigenic gene expression profile.

• MeSH
• adenokarcinom diagnóza   metabolismus   MeSH
• adhezní molekula epiteliálních buněk metabolismus   MeSH
• antigen AC133 metabolismus   MeSH
• antigen CD24 metabolismus   MeSH
• antigeny CD44 metabolismus   MeSH
• kmenové buňky metabolismus   MeSH
• kultivované buňky MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové biomarkery MeSH
• nádory slinivky břišní diagnóza   metabolismus   MeSH
• nestin metabolismus   MeSH
• prognóza MeSH
• průtoková cytometrie MeSH
• regulace genové exprese u nádorů MeSH
• senioři MeSH
• transkriptom MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH