The FGF signaling pathway plays an important role in the regulation of limb development, controlling cell migration, proliferation, differentiation, and apoptosis. Sprouty proteins act as antagonists of the FGF pathway and control the extent of FGF signaling as part of a negative feedback loop. Sprouty2/4 deficient mice evince defects in endochondral bone formation and digit patterning in their forelimbs, with pathogenesis recently related to ciliopathies. To understand the mechanisms behind these pathologies, the limb defects in Sprouty2+/-;Sprouty4-/- male and female mice were characterized and correlated to the dynamic expression patterns of Sprouty2 and Sprouty4, and the impact on the main signaling centers of the limb bud was assessed. Sprouty2 and Sprouty4 exhibited dynamic expressions during limb development. Interestingly, despite similar expression patterns in all limbs, the hindlimbs did not evince any obvious alterations in development, while the forelimbs showed consistent phenotypes of variable severity. Prenatally as well as postnatally, the left forelimb was significantly more severely affected than the right one. A broad variety of pathologies was present in the autopodium of the forelimb, including changes in digit number, size, shape, and number of bones, hand clefts, and digit fusions. Ectopic ossification of bones and abnormal bone fusions detected in micro-CT scans were frequently observed in the digital as well as in the carpal and metacarpal areas. Sprouty2+/-;Sprouty4-/- limb buds showed patchy loss of Fgf8 expression in the apical ectodermal ridge, and a loss of tissue underlying these regions. The zone of polarizing activity was also impacted, with lineage analysis highlighting a change in the contribution of Sonic hedgehog expressing cells. These findings support the link between Sproutys and Hedgehog signaling during limb development and highlight the importance of Sprouty2 and Sprouty4 in controlling early signaling centers in the limb.

• Publikační typ
• časopisecké články MeSH

Tooth ankyloses are pathological conditions in human, which arise by direct fusion of the tooth with surrounding tissues as a result of trauma or inflammation around teeth. Failure of interaction between these tissues lead to the disruption of tooth function as an organ and the resorption of alveolar bone. This state is followed by loss of teeth and by the initiation of conditions inappropriate for following teeth reconstruction. Aim of the project is to uncover cellular and molecular processes contributing to morphological and functional changes during tooth ankyloses initiation in human. Project will use new methodical approaches such as LIBS, micro CT examination or gene expression analyses of osteogenic factors in periodontal area with aim to expand our understanding about causes of teeth ankyloses commencement as well as possibilities of new approaches for their prevention during teeth traumas.

Zubní ankylóza je u člověka patologický proces, který vzniká přímou fúzí zubní tkáně s okolní kostí díky traumatům či zánětu v okolí zubu. Selháním interakce mezi těmito dvěma tkáněmi dochází k poruchám funkce zubu jako orgánu, která může vést k resorpci alveolární kosti. Tento stav končí často ztrátou zubu a vznikem podmínek nevhodných pro následnou rekonstrukci ztraceného zubu. Cílem projektu je odhalení buněčných a molekulárních procesů podílejících se na morfologických a funkčních změnách během vzniku zubní ankylózy u člověka. Projekt rovněž využije nové metodické přístupy jako LIBS, mikro CT analýzy či sledování změn exprese osteogenních faktorů v oblasti paradontu s cílem rozšířit znalosti o příčinách vzniku zubních ankylóz, jakož i možnosti preventivních přístupů při traumatech zubů.

• Klíčová slova
• ankylóza, ankyloses, zub, tooth, osteogenní markery, alveolární kost, LIBS, PCR Array, mikro CT, osteogenetic markers, alveolar bone, LIBS, PCR Array, micro CT,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Infections affecting the root canal of the tooth are the most common cause of apical periodontitis (AP). Chronic inflammation in the periodontium may lead to the development of a radicular cyst (RC), a type of inflammatory odontogenic cyst (OC). OC of developmental origin also includes dentigerous cysts (DC) and odontogenic keratocysts (OKC). Correct classification of OCs reflecting their etiopathogenesis including genetic factors is crucial for choosing an optimum therapy and patient management. The aim of the project is to identify risk factors of OC development and improve their diagnosis. The project includes study of microbiome present in the root canal, genetic association studies focused on patients ́immune profiling and influence of genes envolved in odontogenesis, comparative studies of gene expression profiles in various types of odontogenic cysts, and study of selected molecular pathways in OC pathogenesis in an animal model. Based on the obtained results we will propose a panel of markers for prediction of OC development and differential diagnosis of OC types.

Infekce postihující kořenový kanálek zubu je nejčastější příčinou vzniku apikální periodontitidy (AP). Chronický zánět v periodonciu může vést k rozvoji radikulární cysty (RC), která patří k zánětlivým odontogenním cystám (OC). Další skupinu tvoří OC vývojové, k nimž se mimojiné řadí cysta folikulární (FC) a odontogenní keratocysta (OKC). Pro optimální volbu terapie a manažment pacientů je klasifikace OC, která reflektuje jejich etiopatogenezi a zahrnuje genetické faktory, zásadní. Cílem projektu je determinace rizikových faktorů rozvoje OC a zlepšení jejich diagnostiky. Projekt zahrnuje studii mikrobiomu kořenového kanálku zubu a OC zánětlivého původu, genetické asociační studie se zaměřením na imunitní profil pacientů a geny zapojené do odontogeneze, komparativní studie expresních profilů genů v různých typech OC, a studie vybraných molekulárních drah v patogenezi OC na animálním modelu. Na základě získaných výsledků se pokusíme navrhnout panel markerů pro predikci vzniku OC a jejich diferenciální diagnostiku.

• Klíčová slova
• Genetická predispozice, Genetic predisposition, sequencing, Nanočástice, imunogenetika, nanoparticle, sekvenování, immunogenetics, apical periodontitis, orální mikrobiom, oral microbiome, SHH signalizace, in vivo animální model, odontogenní cysta, apikální periodontitida, SHH signaling, in vivo animal model, odontogenic cyst,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Squamous cell carcinoma (SCC) of the head and neck originates from the mucosal lining of the upper aerodigestive tract, including the lip, tongue, nasopharynx, oropharynx, larynx and hypopharynx. In this review, we summarise what is currently known about the potential function of primary cilia in the pathogenesis of this disease. As primary cilia represent a key cellular structure for signal transduction and are related to cell proliferation, an understanding of their role in carcinogenesis is necessary for the design of new treatment approaches. Here, we introduce cilia-related signalling in head and neck squamous cell carcinoma (HNSCC) and its possible association with HNSCC tumorigenesis. From this point of view, PDGF, EGF, Wnt and Hh signalling are discussed as all these pathways were found to be dysregulated in HNSCC. Moreover, we review the clinical potential of small molecules affecting primary cilia signalling to target squamous cell carcinoma of the head and neck area.

• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

CDK13-related disorder, also known as congenital heart defects, dysmorphic facial features and intellectual developmental disorder (CHDFIDD) is associated with mutations in the CDK13 gene encoding transcription-regulating cyclin-dependent kinase 13 (CDK13). Here, we focused on the development of craniofacial structures and analyzed early embryonic stages in CHDFIDD mouse models, with one model comprising a hypomorphic mutation in Cdk13 and exhibiting cleft lip/palate, and another model comprising knockout of Cdk13, featuring a stronger phenotype including midfacial cleft. Cdk13 was found to be physiologically expressed at high levels in the mouse embryonic craniofacial structures, namely in the forebrain, nasal epithelium and maxillary mesenchyme. We also uncovered that Cdk13 deficiency leads to development of hypoplastic branches of the trigeminal nerve including the maxillary branch. Additionally, we detected significant changes in the expression levels of genes involved in neurogenesis (Ache, Dcx, Mef2c, Neurog1, Ntn1, Pou4f1) within the developing palatal shelves. These results, together with changes in the expression pattern of other key face-specific genes (Fgf8, Foxd1, Msx1, Meis2 and Shh) at early stages in Cdk13 mutant embryos, demonstrate a key role of CDK13 in the regulation of craniofacial morphogenesis.

• MeSH
• cyklin-dependentní kinasy metabolismus   genetika   MeSH
• embryo savčí metabolismus   MeSH
• embryonální vývoj * genetika   MeSH
• fenotyp MeSH
• lebka embryologie   patologie   MeSH
• mentální retardace genetika   MeSH
• modely nemocí na zvířatech * MeSH
• mutace genetika   MeSH
• myši MeSH
• nervus trigeminus embryologie   MeSH
• neurogeneze * genetika   MeSH
• obličej embryologie   abnormality   MeSH
• protein doublecortin MeSH
• rozštěp patra genetika   patologie   embryologie   MeSH
• rozštěp rtu genetika   patologie   embryologie   MeSH
• vývojová regulace genové exprese * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVE: Oral squamous cell carcinoma (OSCC) originates from the mucosal lining of the oral cavity. Almost half of newly diagnosed cases are classified as advanced stage IV disease, which makes resection difficult. In this study, we investigated the pathological features and mutation profiles of tumor margins in OSCC. METHODS: We performed hierarchical clustering of principal components to identify distinct patterns of tumor growth and their association with patient prognosis. We also used next-generation sequencing to analyze somatic mutations in tumor and marginal tissue samples. RESULTS: Our analyses uncovered that the grade of worst pattern of invasion (WPOI) is strongly associated with depth of invasion and patient survival in multivariable analysis. Mutations were primarily detected in the DNA isolated from tumors, but several mutations were also identified in marginal tissue. In total, we uncovered 29 mutated genes, mainly tumor suppressor genes involved in DNA repair including BRCA genes; however none of these mutations significantly correlated with a higher chance of relapse in our medium-size cohort. Some resection margins that appeared histologically normal harbored tumorigenic mutations in TP53 and CDKN2A genes. CONCLUSION: Even histologically normal margins may contain molecular alterations that are not detectable by conventional histopathological methods, but NCCN classification system still outperforms other methods in the prediction of the probability of disease relapse.

• Publikační typ
• časopisecké články MeSH

• MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• myši knockoutované MeSH
• prořezávání zubů MeSH
• úrazy zubů komplikace   MeSH
• zubní ankylóza * diagnóza   etiologie   terapie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH

Primary cilia are cellular surface projections enriched in receptors and signaling molecules, acting as signaling hubs that respond to stimuli. Malfunctions in primary cilia have been linked to human diseases, including retinopathies and ocular defects. Here, we focus on TMEM107, a protein localized to the transition zone of primary cilia. TMEM107 mutations were found in patients with Joubert and Meckel-Gruber syndromes. A mouse model lacking Tmem107 exhibited eye defects such as anophthalmia and microphthalmia, affecting retina differentiation. Tmem107 expression during prenatal mouse development correlated with phenotype occurrence, with enhanced expression in differentiating retina and optic stalk. TMEM107 deficiency in retinal organoids resulted in the loss of primary cilia, down-regulation of retina-specific genes, and cyst formation. Knocking out TMEM107 in human ARPE-19 cells prevented primary cilia formation and impaired response to Smoothened agonist treatment because of ectopic activation of the SHH pathway. Our data suggest TMEM107 plays a crucial role in early vertebrate eye development and ciliogenesis in the differentiating retina.

• MeSH
• lidé MeSH
• membránové proteiny genetika   metabolismus   MeSH
• myši MeSH
• polycystická choroba ledvin * genetika   MeSH
• poruchy ciliární motility * genetika   metabolismus   MeSH
• retina metabolismus   MeSH
• retinopathia pigmentosa * metabolismus   MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Carcinomas of the oral cavity and oropharynx belong among the ten most common malignancies in human population. In this project, we will focus on the most common squamous cell carcinoma. The prognosis of these malignancies is determined by the degree of invasiveness of the primary tumor and by the extent of metastatic spread into regional and distant lymph nodes. The intensity of perineural invasion correlates with tumor localization, its extent, and the presence of nodal metastases. In our project, we will focus on the detection of clinically relevant somatic mutations in patients with perineural invasion and the evaluation of early molecular markers during perineural invasion, which are expressed on the interface between tumor and surrounding tissues. Moreover, we will analyze role of primary cilia and Sonic hedgehog pathway in tumorous cells and the effect of altered SHH signaling on perineural invasion. Our aim is to introduce new protocols into routine histopathological diagnostics aiming to predict future cell behavior, which will improve planning and management of the subsequent therapies in patients.

Karcinomy dutiny ústní a orofaryngu patří mezi deset nejčastěji se vyskytujících malignit v lidské populaci. Náš projekt je zaměřen na spinocelulární karcinom, který představuje v této oblasti nejfrekventovanější typ maligního onemocnění. Prognóza onemocnění zhoubným nádorem dutiny ústní a orofaryngu je dána především stupněm invazivity primárního tumoru a rozsahem metastatického postižení regionálních a vzdálených uzlin. Intenzita perineurální invaze koreluje s lokalizací nádoru, jeho rozsahem a přítomností uzlinových metastáz. V našem projektu se zaměříme na determinaci klinicky relevantních somatických mutací u pacientů s perineurální invazí a na analýzu exprese raných molekulárních markerů perineurální invaze na rozhraní nádorové a okolní tkáně. Dále budeme analyzovat úlohu primárních cilií a Sonic Hedgehog dráhy v buňkách spinocelulárního karcinomu a vliv modifikované SHH signalizace na jejich perineurální invazi. Primárním cílem projektu je zavést do rutinní histopatologické diagnostiky nové postupy umožňující lépe predikovat budoucí chování nádorových buněk, které povede ke zlepšení plánování další terapie u pacientů.

• Klíčová slova
• karcinom, carcinoma, dutina ústní, orofarynx, perineurální invaze, oral cavity, oropharynx, predictive factor, perineural invasion, prognostický marker, prognostic marker, predictivní faktor,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

BACKGROUND: Ameloblastic carcinoma and metastasising ameloblastoma are rare epithelial odontogenic tumours with aggressive features. Distinguishing between these two lesions is often clinically difficult but necessary to predict tumour behaviour or to plan future therapy. Here, we provide a brief review of the literature available on these two types of lesions and present a new case report of a young man with an ameloblastoma displaying metastatic features. We also use this case to illustrate the similarities and differences between these two types of tumours and the difficulties of their differential diagnosis. CASE PRESENTATION: Our histopathological analyses uncovered a metastasising tumour with features of ameloblastic carcinoma, which developed from the ameloblastoma. We profiled the gene expression of Wnt pathway members in ameloblastoma sample of this patient, because multiple molecules of this pathway are involved in the establishing of cell polarity, cell migration or for epithelial-mesenchymal transition during tumour metastasis to evaluate features of tumor behaviour. Indeed, we found upregulation of several cell migration-related genes in our patient. Moreover, we uncovered somatic mutation BRAF p.V600E with known pathological role in cancerogenesis and germline heterozygous FANCA p.S858R mutation, whose interpretation in this context has not been discussed yet. CONCLUSIONS: In conclusion, we have uncovered a unique case of ameloblastic carcinoma associated with an alteration of Wnt signalling and the presence of BRAF mutation. Development of harmful state of our patient might be also supported by the germline mutation in one FANCA allele, however this has to be confirmed by further analyses.

• MeSH
• ameloblastom * genetika   diagnóza   MeSH
• karcinom * patologie   MeSH
• lidé MeSH
• mutace MeSH
• odontogenní nádory * diagnóza   genetika   MeSH
• protoonkogenní proteiny B-raf genetika   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• přehledy MeSH