BACKGROUND: Fuchs endothelial corneal dystrophy (FECD) is the most common repeat-mediated disease in humans. It exclusively affects corneal endothelial cells (CECs), with ≤81% of cases associated with an intronic TCF4 triplet repeat (CTG18.1). Here, we utilise optical genome mapping (OGM) to investigate CTG18.1 tissue-specific instability to gain mechanistic insights. METHODS: We applied OGM to a diverse range of genomic DNAs (gDNAs) from patients with FECD and controls (n = 43); CECs, leukocytes and fibroblasts. A bioinformatics pipeline was developed to robustly interrogate CTG18.1-spanning DNA molecules. All results were compared with conventional polymerase chain reaction-based fragment analysis. FINDINGS: Analysis of bio-samples revealed that expanded CTG18.1 alleles behave dynamically, regardless of cell-type origin. However, clusters of CTG18.1 molecules, encompassing ∼1800-11,900 repeats, were exclusively detected in diseased CECs from expansion-positive cases. Additionally, both progenitor allele size and age were found to influence the level of leukocyte-specific CTG18.1 instability. INTERPRETATION: OGM is a powerful tool for analysing somatic instability of repeat loci and reveals here the extreme levels of CTG18.1 instability occurring within diseased CECs underpinning FECD pathophysiology, opening up new therapeutic avenues for FECD. Furthermore, these findings highlight the broader translational utility of FECD as a model for developing therapeutic strategies for rarer diseases similarly attributed to somatically unstable repeats. FUNDING: UK Research and Innovation, Moorfields Eye Charity, Fight for Sight, Medical Research Council, NIHR BRC at Moorfields Eye Hospital and UCL Institute of Ophthalmology, Grantová Agentura České Republiky, Univerzita Karlova v Praze, the National Brain Appeal's Innovation Fund and Rosetrees Trust.

• Klíčová slova
• Fuchs endothelial corneal dystrophy, Optical genome mapping, Somatic mosaicism, Tissue-specific repeat instability, Trinucleotide repeat expansion disease, Triplet repeat expansion-mediated disease,
• MeSH
• alely MeSH
• expanze trinukleotidových repetic MeSH
• Fuchsova endoteliální dystrofie * genetika   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mapování chromozomů MeSH
• nestabilita genomu MeSH
• orgánová specificita genetika   MeSH
• senioři MeSH
• transkripční faktor 4 * genetika   metabolismus   MeSH
• trinukleotidové repetice genetika   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• TCF4 protein, human MeSH Prohlížeč
• transkripční faktor 4 * MeSH

BACKGROUND: Slow neurotransmission including DARPP-32 signalling is implicated in substance use disorders (SUDs) by experimental systems but not yet in the human aetiology. PPP1R12B, encoding another protein in the DARPP-32 family, hasn't been studied in the brain. METHODS: Brain-regional gene activity was assessed in three different animal models of SUDs for mRNA level alterations. Genetic associations were assessed by meta-analysis of pre-existing dbGaP GWAS datasets for main effects and epistasis with known genetic risks, followed by cell type-specific pathway delineation. Parkinson's disease (PD) was included as a dopamine-related disease control for SUDs. FINDINGS: In animal models of SUDs, environmentally-altered PPP1R12B expression sex-dependently involves motivation-related brain regions. In humans with polysubstance abuse, meta-analysis of pre-existing datasets revealed that PPP1R12B and PPP1R1B, although expressed in dopamine vs. dopamine-recipient neurons, exerted similar interactions with known genetic risks such as ACTR1B and DRD2 in men but with ADH1B, HGFAC and DRD3 in women. These interactions reached genome-wide significances (Pmeta<10-20) for SUDs but not for PD (disease selectivity: P = 4.8 × 10-142, OR = 6.7 for PPP1R12B; P = 8.0 × 10-8, OR = 2.1 for PPP1R1B). CADM2 was the common risk in the molecular signalling regardless of gender and cell type. INTERPRETATION: Gender-dependant slow neurotransmission may convey both genetic and environmental vulnerabilities selectively to SUDs. FUNDING: Grants from National Institute on Drug Abuse (NIDA) and National Institute on Alcohol Abuse and Alcoholism (NIAAA) of U.S.A. and National Natural Science Foundation of China (NSFC).

• Klíčová slova
• Adolescence, Cell type-specific, Environmental risk, Missing heritability, Polysubstance abuse, Slow neurotransmission,
• MeSH
• celogenomová asociační studie MeSH
• dopaminem a cAMP regulovaný fosfoprotein 32 genetika   metabolismus   MeSH
• genetická epistáze * MeSH
• genetická heterogenita MeSH
• genetická predispozice k nemoci MeSH
• genové regulační sítě MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• mozek metabolismus   patologie   MeSH
• myši MeSH
• náchylnost k nemoci MeSH
• nervový přenos genetika   MeSH
• orgánová specificita genetika   MeSH
• poruchy spojené s užíváním psychoaktivních látek diagnóza   etiologie   metabolismus   MeSH
• proteinfosfatasa 1 genetika   metabolismus   MeSH
• regulace genové exprese MeSH
• sexuální faktory MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• dopaminem a cAMP regulovaný fosfoprotein 32 MeSH
• PPP1R12B protein, human MeSH Prohlížeč
• PPP1R1B protein, human MeSH Prohlížeč
• proteinfosfatasa 1 MeSH

In recent years, a lot of scientific interest has focused on cancer immunotherapy. Although chronic inflammation has been described as one of the hallmarks of cancer, acute inflammation can actually trigger the immune system to fight diseases, including cancer. Toll-like receptor (TLR) ligands have long been used as adjuvants for traditional vaccines and it seems they may also play a role enhancing efficiency of tumor immunotherapy. The aim of this perspective is to discuss the effects of TLR stimulation in cancer, expression of various TLRs in different types of tumors, and finally the role of TLRs in anti-cancer immunity and tumor rejection.

• Klíčová slova
• anti-cancer immunity, immuno-oncology, immunotherapy, toll-like receptors, tumor rejection,
• MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• imunita * MeSH
• lidé MeSH
• ligandy MeSH
• nádory etiologie   metabolismus   patologie   terapie   MeSH
• orgánová specificita genetika   imunologie   MeSH
• regulace genové exprese u nádorů MeSH
• toll-like receptory agonisté   genetika   metabolismus   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• ligandy MeSH
• toll-like receptory MeSH

The physiological function of the pancreas is controlled by the circadian clock. The aim of this study was to determine whether aging-induced changes in glucose homeostasis affect properties of the circadian clock in the pancreas and/or its sensitivity to disturbances in environmental lighting conditions. mPer2Luc mice aged 24-26 months developed hyperinsulinemic hypoglycaemia, which was likely due to the Pclo-mediated insulin hyper-secretion and Slc2a2-mediated glucose transport impairment in the pancreas, and due to the alterations in Pp1r3c-related glycogen storage and Sgk1-related glucose transport in the liver. In the pancreatic tissue, aging affected clock gene expression only marginally, it upregulated Bmal1 and downregulated Clock expression. Whereas aging significantly impaired the circadian clock in lung explants, which were used as a control tissue, the properties of the pancreatic clock in vitro were not affected. The data suggest a non-circadian role of Bmal1 in changes of pancreatic function that occur during aging. Additionally, the pancreatic clock was more sensitive to exposure of animals to constant light conditions. These findings provide an explanation for the previously demonstrated relationship between disturbances in the circadian system and disordered glucose homeostasis, including diabetes mellitus type 2, in subjects exposed to long-term shift work.

• MeSH
• cirkadiánní hodiny * účinky záření   MeSH
• cirkadiánní proteiny Period metabolismus   MeSH
• glukosa metabolismus   MeSH
• homeostáza * MeSH
• játra metabolismus   MeSH
• kolon metabolismus   MeSH
• myši MeSH
• orgánová specificita genetika   MeSH
• pankreas metabolismus   účinky záření   MeSH
• proteiny CLOCK genetika   metabolismus   MeSH
• regulace genové exprese účinky záření   MeSH
• stárnutí metabolismus   MeSH
• světlo MeSH
• transkripční faktory ARNTL genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Bmal1 protein, mouse MeSH Prohlížeč
• cirkadiánní proteiny Period MeSH
• glukosa MeSH
• Per2 protein, mouse MeSH Prohlížeč
• proteiny CLOCK MeSH
• transkripční faktory ARNTL MeSH

Adenosine-to-inosine (A-to-I) RNA editing is a conserved post-transcriptional mechanism mediated by ADAR enzymes that diversifies the transcriptome by altering selected nucleotides in RNA molecules. Although many editing sites have recently been discovered, the extent to which most sites are edited and how the editing is regulated in different biological contexts are not fully understood. Here we report dynamic spatiotemporal patterns and new regulators of RNA editing, discovered through an extensive profiling of A-to-I RNA editing in 8,551 human samples (representing 53 body sites from 552 individuals) from the Genotype-Tissue Expression (GTEx) project and in hundreds of other primate and mouse samples. We show that editing levels in non-repetitive coding regions vary more between tissues than editing levels in repetitive regions. Globally, ADAR1 is the primary editor of repetitive sites and ADAR2 is the primary editor of non-repetitive coding sites, whereas the catalytically inactive ADAR3 predominantly acts as an inhibitor of editing. Cross-species analysis of RNA editing in several tissues revealed that species, rather than tissue type, is the primary determinant of editing levels, suggesting stronger cis-directed regulation of RNA editing for most sites, although the small set of conserved coding sites is under stronger trans-regulation. In addition, we curated an extensive set of ADAR1 and ADAR2 targets and showed that many editing sites display distinct tissue-specific regulation by the ADAR enzymes in vivo. Further analysis of the GTEx data revealed several potential regulators of editing, such as AIMP2, which reduces editing in muscles by enhancing the degradation of the ADAR proteins. Collectively, our work provides insights into the complex cis- and trans-regulation of A-to-I editing.

• MeSH
• adenosindeaminasa * genetika   metabolismus   MeSH
• časoprostorová analýza MeSH
• druhová specificita MeSH
• editace RNA genetika   MeSH
• genotyp MeSH
• HEK293 buňky MeSH
• jaderné proteiny metabolismus   MeSH
• lidé MeSH
• myši MeSH
• orgánová specificita genetika   MeSH
• primáti genetika   MeSH
• proteiny vázající RNA * genetika   metabolismus   MeSH
• proteolýza MeSH
• svaly metabolismus   MeSH
• transkriptom genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• ADAR protein, human MeSH Prohlížeč
• ADAR1 protein, mouse MeSH Prohlížeč
• ADAR2 protein, mouse MeSH Prohlížeč
• ADARB1 protein, human MeSH Prohlížeč
• adenosindeaminasa * MeSH
• AIMP2 protein, human MeSH Prohlížeč
• jaderné proteiny MeSH
• proteiny vázající RNA * MeSH

In tobacco, three sequence variants of the TERT gene have been described. We revealed unbalanced levels of TERT variant transcripts in vegetative tobacco tissues and enhanced TERT transcription and telomerase activity in reproductive tissues. Telomerase is a ribonucleoprotein complex responsible for the maintenance of telomeres, structures delimiting ends of linear eukaryotic chromosomes. In the Nicotiana tabacum (tobacco) allotetraploid plant, three sequence variants (paralogs) of the gene coding for the telomerase reverse transcriptase subunit (TERT) have been described, two of them derived from the maternal N. sylvestris genome (TERT_Cs, TERT_D) and one originated from the N. tomentosiformis paternal genome (TERT_Ct). In this work, we analyzed the transcription of TERT variants in correlation with telomerase activity in tobacco tissues. High and approximately comparable levels of TERT_Ct and TERT_Cs transcripts were detected in seedlings, roots, flower buds and leaves, while the transcript of the TERT_D variant was markedly underrepresented. Similarly, in N. sylvestris tissues, TERT_Cs transcript significantly predominated. A specific pattern of TERT transcripts was found in samples of tobacco pollen with the TERT_Cs variant clearly dominating particularly at the early stage of pollen development. Detailed analysis of TERT_C variants representation in functionally distinct fractions of pollen transcriptome revealed their prevalence in large ribonucleoprotein particles encompassing translationally silent mRNA; only a minority of TERT_Ct and TERT_Cs transcripts were localized in actively translated polysomes. Histones of the TERT_C chromatin were decorated predominantly with the euchromatin-specific epigenetic modification in both telomerase-positive and telomerase-negative tobacco tissues. We conclude that the existence and transcription pattern of tobacco TERT paralogs represents an interesting phenomenon and our results indicate its functional significance. Nicotiana species have again proved to be appropriate and useful model plants in telomere biology studies.

• Klíčová slova
• Gene sequence variant, Pollen, Polyploids, Telomerase, Telomere, Transcription,
• MeSH
• buněčné jádro genetika   MeSH
• chromatinová imunoprecipitace MeSH
• euchromatin metabolismus   MeSH
• genetická transkripce MeSH
• genetická variace * MeSH
• histony metabolismus   MeSH
• messenger RNA genetika   metabolismus   MeSH
• orgánová specificita genetika   MeSH
• polyribozomy metabolismus   MeSH
• posttranslační úpravy proteinů MeSH
• pylová láčka růst a vývoj   MeSH
• regulace genové exprese u rostlin * MeSH
• tabák genetika   MeSH
• telomerasa genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• euchromatin MeSH
• histony MeSH
• messenger RNA MeSH
• telomerasa MeSH

Cephalochordates, the sister group of vertebrates + tunicates, are evolving particularly slowly. Therefore, genome comparisons between two congeners of Branchiostoma revealed so many conserved noncoding elements (CNEs), that it was not clear how many are functional regulatory elements. To more effectively identify CNEs with potential regulatory functions, we compared noncoding sequences of genomes of the most phylogenetically distant cephalochordate genera, Asymmetron and Branchiostoma, which diverged approximately 120-160 million years ago. We found 113,070 noncoding elements conserved between the two species, amounting to 3.3% of the genome. The genomic distribution, target gene ontology, and enriched motifs of these CNEs all suggest that many of them are probably cis-regulatory elements. More than 90% of previously verified amphioxus regulatory elements were re-captured in this study. A search of the cephalochordate CNEs around 50 developmental genes in several vertebrate genomes revealed eight CNEs conserved between cephalochordates and vertebrates, indicating sequence conservation over >500 million years of divergence. The function of five CNEs was tested in reporter assays in zebrafish, and one was also tested in amphioxus. All five CNEs proved to be tissue-specific enhancers. Taken together, these findings indicate that even though Branchiostoma and Asymmetron are distantly related, as they are evolving slowly, comparisons between them are likely optimal for identifying most of their tissue-specific cis-regulatory elements laying the foundation for functional characterizations and a better understanding of the evolution of developmental regulation in cephalochordates.

• Klíčová slova
• CNE, amphioxus, asymmetron, cephalochordate, regulatory element,
• MeSH
• bezlebeční genetika   MeSH
• Chordata genetika   MeSH
• fylogeneze MeSH
• genom MeSH
• konzervovaná sekvence genetika   MeSH
• kopinatci genetika   MeSH
• molekulární evoluce * MeSH
• orgánová specificita genetika   MeSH
• regulační oblasti nukleových kyselin genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

Ixodes ricinus is a tick that transmits the pathogens of Lyme and several arboviral diseases. Pathogens invade the tick midgut, disseminate through the hemolymph, and are transmitted to the vertebrate host via the salivary glands; subverting these processes could be used to interrupt pathogen transfer. Here, we use massive de novo sequencing to characterize the transcriptional dynamics of the salivary and midgut tissues of nymphal and adult I. ricinus at various time points after attachment on the vertebrate host. Members of a number of gene families show stage- and time-specific expression. We hypothesize that gene expression switching may be under epigenetic control and, in support of this, identify 34 candidate proteins that modify histones. I. ricinus-secreted proteins are encoded by genes that have a non-synonymous to synonymous mutation rate even greater than immune-related genes. Midgut transcriptome (mialome) analysis reveals several enzymes associated with protein, carbohydrate, and lipid digestion, transporters and channels that might be associated with nutrient uptake, and immune-related transcripts including antimicrobial peptides. This publicly available dataset supports the identification of protein and gene targets for biochemical and physiological studies that exploit the transmission lifecycle of this disease vector for preventative and therapeutic purposes.

• MeSH
• anotace sekvence MeSH
• časové faktory MeSH
• fylogeneze MeSH
• genetická transkripce * MeSH
• klíště klasifikace   genetika   MeSH
• orgánová specificita genetika   MeSH
• polymorfismus genetický MeSH
• shluková analýza MeSH
• slinné žlázy metabolismus   MeSH
• stanovení celkové genové exprese MeSH
• střevní sliznice metabolismus   MeSH
• transkriptom MeSH
• výpočetní biologie MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Intramural MeSH

Nuclear receptors (NRs), or nuclear hormone receptors (NHRs), are transcription factors that regulate development and metabolism of most if not all animal species. Their regulatory networks include conserved mechanisms that are shared in-between species as well as mechanisms that are restricted to certain phyla or even species. In search for conserved members of the NHR family in Schmidtea mediterranea, we identified a molecular signature of a class of NRs, NR2E1, in the S. mediterranea genome and cloned its complete cDNA coding sequence. The derived amino acid sequence shows a high degree of conservation of both DNA-binding domain and ligand- binding domain and a remarkably high homology to vertebrate NR2E1 and C. elegans NHR-67. Quantitative PCR detected approximately ten-fold higher expression of Smed-tlx-1 in the proximal part of the head compared to the tail region. The expression of Smed-tlx-1 is higher during fed state than during fasting. Smed-tlx-1 down-regulation by RNA interference affects the ability of the animals to maintain body plan and induces defects of brain, eyes and body shape during fasting and re-growing cycles. These results suggest that SMED-TLX-1 is critical for tissue and body plan maintenance in planaria.

• MeSH
• fylogeneze MeSH
• klonování DNA MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• omezení příjmu potravy fyziologie   MeSH
• orgánová specificita genetika   MeSH
• proteiny červů chemie   genetika   metabolismus   MeSH
• receptory cytoplazmatické a nukleární chemie   genetika   metabolismus   MeSH
• regulace genové exprese MeSH
• RNA interference MeSH
• rozvržení tělního plánu * genetika   MeSH
• sekvence aminokyselin MeSH
• sekvenční seřazení MeSH
• stravovací zvyklosti fyziologie   MeSH
• Turbellaria embryologie   genetika   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• proteiny červů MeSH
• receptory cytoplazmatické a nukleární MeSH

Hemojuvelin (Hjv) is an essential component of the pathway regulating hepcidin (Hamp1) gene expression. Mice with targeted disruption of the Hjv gene (Hjv-/- mice) fail to upregulate hepatic Hamp1 expression following iron overload. The main aim of the study was to determine whether the Hjv protein is also necessary for Hamp1 downregulation. In addition, sex differences in Hamp1 expression in Hjv-/- mice were also examined. Male and female Hjv-/- mice (129SvJ background) were used for the experiments, tissue Hamp1 and Hamp2 mRNA content was determined by real-time PCR. Hepatic Hamp1 mRNA content in male Hjv-/- mice was low (0.6% of Hjv+/+ males), however, female Hjv-/- mice displayed only moderately reduced (to 17%) Hamp1 mRNA levels. Hepatic non-heme iron concentration was similar in Hjv-/- mice of both sexes. Disruption of the Hjv gene did not affect Hamp1 mRNA content in the myocardium or Hamp2 mRNA content in the pancreas. Single phlebotomy resulted in significant reduction of Hamp1 mRNA in both male and female Hjv+/+ mice (to 17% and 27% of controls respectively), measured 20 h after treatment. In Hjv-/- mice, phlebotomy decreased Hamp1 mRNA content to 46% in males and to 11% in females. Bleeding also significantly decreased (to 16%) hepatic Hamp2 mRNA levels in Hjv-/- females. The obtained results indicate that the pathway mediating hepcidin downregulation by phlebotomy does not require functional hemojuvelin protein. In addition, they confirm a significant effect of sex on hepcidin gene expression.

• MeSH
• flebotomie MeSH
• GPI-vázané proteiny MeSH
• hepcidiny MeSH
• kationické antimikrobiální peptidy biosyntéza   MeSH
• krvácení metabolismus   MeSH
• lidé MeSH
• membránové proteiny nedostatek   MeSH
• mutace MeSH
• myši knockoutované MeSH
• myši MeSH
• orgánová specificita genetika   MeSH
• pohlavní dimorfismus * MeSH
• protein hemochromatózy MeSH
• regulace genové exprese * genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• GPI-vázané proteiny MeSH
• HAMP protein, human MeSH Prohlížeč
• Hamp protein, mouse MeSH Prohlížeč
• Hamp2 protein, mouse MeSH Prohlížeč
• hepcidiny MeSH
• HJV protein, mouse MeSH Prohlížeč
• kationické antimikrobiální peptidy MeSH
• membránové proteiny MeSH
• protein hemochromatózy MeSH