OBJECTIVES: Hedgehog signalling plays a critical role during the pathogenesis of fibrosis in systemic sclerosis (SSc). Besides canonical hedgehog signalling with smoothened (SMO)-dependent activation of GLI transcription factors, GLI can be activated independently of classical hedgehog ligands and receptors (so-called non-canonical pathways). Here, we aimed to evaluate the role of non-canonical hedgehog signalling in SSc and to test the efficacy of direct GLI inhibitors that target simultaneously canonical and non-canonical hedgehog pathways. METHODS: The GLI inhibitor GANT-61 was used to inhibit canonical as well as non-canonical hedgehog signalling, while the SMO inhibitor vismodegib was used to selectively target canonical hedgehog signalling. Furthermore, GLI2 was selectively depleted in fibroblasts using the Cre-LoxP system. The effects of pharmacological or genetic of GLI2 on transforming growth factor-β (TGF-β) signalling were analysed in cultured fibroblasts, in bleomycin-induced pulmonary fibrosis and in mice with overexpression of a constitutively active TGF-β receptor I. RESULTS: TGF-β upregulated GLI2 in a Smad3-dependent manner and induced nuclear accumulation and DNA binding of GLI2. Fibroblast-specific knockout of GLI2 protected mice from TBRact-induced fibrosis. Combined targeting of canonical and non-canonical hedgehog signalling with direct GLI inhibitors exerted more potent antifibrotic effects than selective targeting of canonical hedgehog signalling with SMO inhibitors in experimental dermal and pulmonary fibrosis. CONCLUSIONS: Our data demonstrate that hedgehog pathways and TGF-β signalling both converge to GLI2 and that GLI2 integrates those signalling to promote tissue fibrosis. These findings may have translational implications as non-selective inhibitors of GLI2 are in clinical use and selective molecules are currently in development.

• Klíčová slova
• Fibroblasts, Systemic Sclerosis, Treatment,
• MeSH
• anilidy farmakologie   MeSH
• dospělí MeSH
• fibroblasty účinky léků   metabolismus   MeSH
• fibróza MeSH
• genový knockout MeSH
• inhibitor aktivátoru plazminogenu 1 genetika   MeSH
• kolagen typu I genetika   MeSH
• kultivované buňky MeSH
• kůže účinky léků   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• mladý dospělý MeSH
• myši knockoutované MeSH
• myši transgenní MeSH
• myši MeSH
• plicní fibróza chemicky indukované   metabolismus   MeSH
• protein Gli2 s motivem zinkových prstů MeSH
• protein Smad3 metabolismus   MeSH
• protein-serin-threoninkinasy antagonisté a inhibitory   genetika   MeSH
• proteiny hedgehog metabolismus   MeSH
• pteridiny farmakologie   MeSH
• pyridiny farmakologie   MeSH
• pyrimidiny farmakologie   MeSH
• receptor Smoothened antagonisté a inhibitory   MeSH
• receptory transformujícího růstového faktoru beta antagonisté a inhibitory   genetika   MeSH
• rekombinantní proteiny farmakologie   MeSH
• růstový faktor pojivové tkáně genetika   MeSH
• senioři MeSH
• signální transdukce účinky léků   MeSH
• systémová sklerodermie genetika   metabolismus   MeSH
• TGF-beta receptor I. typu MeSH
• transformující růstový faktor beta metabolismus   farmakologie   MeSH
• transkripční faktory Krüppel-like antagonisté a inhibitory   genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• myši MeSH
• senioři MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• anilidy MeSH
• GANT 61 MeSH Prohlížeč
• Gli2 protein, mouse MeSH Prohlížeč
• HhAntag691 MeSH Prohlížeč
• inhibitor aktivátoru plazminogenu 1 MeSH
• kolagen typu I MeSH
• messenger RNA MeSH
• protein Gli2 s motivem zinkových prstů MeSH
• protein Smad3 MeSH
• protein-serin-threoninkinasy MeSH
• proteiny hedgehog MeSH
• pteridiny MeSH
• pyridiny MeSH
• pyrimidiny MeSH
• receptor Smoothened MeSH
• receptory transformujícího růstového faktoru beta MeSH
• rekombinantní proteiny MeSH
• růstový faktor pojivové tkáně MeSH
• SD-208 MeSH Prohlížeč
• TGF-beta receptor I. typu MeSH
• transformující růstový faktor beta MeSH
• transkripční faktory Krüppel-like MeSH

The sonic Hedgehog/GLI signaling pathway (HH) is critical for maintaining tissue polarity in development and contributes to tumor stemness. Transcription factors GLI1⁻3 are the downstream effectors of HH and activate oncogenic targets. To explore the completeness of the expression of HH components in tumor cells, we performed a screen for all HH proteins in a wide spectrum of 56 tumor cell lines of various origin using Western blot analysis. Generally, all HH proteins were expressed. Important factors GLI1 and GLI2 were always expressed, only exceptionally one of them was lowered, suggesting the functionality of HH in all tumors tested. We determined the effect of a GLI inhibitor GANT61 on proliferation in 16 chosen cell lines. More than half of tumor cells were sensitive to GANT61 to various extents. GANT61 killed the sensitive cells through apoptosis. The inhibition of reporter activity containing 12xGLI consensus sites by GANT61 and cyclopamine roughly correlated with cell proliferation influenced by GANT61. Our results recognize the sensitivity of tumor cell types to GANT61 in cell culture and support a critical role for GLI factors in tumor progression through restraining apoptosis. The use of GANT61 in combined targeted therapy of sensitive tumors, such as melanomas, seems to be immensely helpful.

• Klíčová slova
• GANT61, GLI, Hedgehog, apoptosis, tumor cell lines,
• MeSH
• HeLa buňky MeSH
• jaderné proteiny metabolismus   MeSH
• Jurkat buňky MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory farmakoterapie   metabolismus   MeSH
• progrese nemoci MeSH
• proliferace buněk účinky léků   MeSH
• protein Gli1 metabolismus   MeSH
• protein Gli2 s motivem zinkových prstů metabolismus   MeSH
• proteiny hedgehog metabolismus   MeSH
• pyridiny farmakologie   MeSH
• pyrimidiny farmakologie   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• signální transdukce účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• GANT 61 MeSH Prohlížeč
• GLI1 protein, human MeSH Prohlížeč
• GLI2 protein, human MeSH Prohlížeč
• jaderné proteiny MeSH
• protein Gli1 MeSH
• protein Gli2 s motivem zinkových prstů MeSH
• proteiny hedgehog MeSH
• pyridiny MeSH
• pyrimidiny MeSH

Microphthalmia-associated transcription factor (MITF) is a nodal point in melanoma transcriptional network that regulates dozens of genes with critical functions in cell differentiation, proliferation and survival. Highly variable MITF expression levels exist in tumor cell subpopulations conferring marked heterogeneity and plasticity in the tumor tissue. A model has been postulated whereby lower MITF levels favour cell invasion and suppress proliferation, whereas high levels stimulate differentiation and proliferation. Additionally, MITF is considered to be a prosurvival gene and a lineage addiction oncogene in melanoma. Herein, we review how MITF expression may affect the melanoma phenotype with consequences on the survival, invasion and metastasis of melanoma cells, and we discuss the research challenges.

• Klíčová slova
• BRN2, GLI2, SOX2, melanoma, microphthalmia-associated transcription factor,
• MeSH
• apoptóza MeSH
• biologické modely MeSH
• epitelo-mezenchymální tranzice MeSH
• exprese genu MeSH
• faktory domény POU metabolismus   MeSH
• homeodoménové proteiny metabolismus   MeSH
• invazivní růst nádoru MeSH
• jaderné proteiny metabolismus   MeSH
• lidé MeSH
• MAP kinasový signální systém MeSH
• melanom genetika   metabolismus   patologie   MeSH
• mikro RNA genetika   metabolismus   MeSH
• nádorové mikroprostředí MeSH
• nádory kůže genetika   metabolismus   patologie   MeSH
• proliferace buněk MeSH
• protein Gli2 s motivem zinkových prstů MeSH
• RNA nádorová genetika   metabolismus   MeSH
• stabilita proteinů MeSH
• transkripční faktor spojený s mikroftalmií genetika   metabolismus   MeSH
• transkripční faktory Krüppel-like metabolismus   MeSH
• transkripční faktory SOXB1 metabolismus   MeSH
• viabilita buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• faktory domény POU MeSH
• GLI2 protein, human MeSH Prohlížeč
• homeodoménové proteiny MeSH
• jaderné proteiny MeSH
• mikro RNA MeSH
• MITF protein, human MeSH Prohlížeč
• protein Gli2 s motivem zinkových prstů MeSH
• RNA nádorová MeSH
• SOX2 protein, human MeSH Prohlížeč
• transcription factor Brn-2 MeSH Prohlížeč
• transkripční faktor spojený s mikroftalmií MeSH
• transkripční faktory Krüppel-like MeSH
• transkripční faktory SOXB1 MeSH

MEK kinase inhibitors (trametinib and selumetinib) or kinase inhibitors directed against mutated BRAF(V600E) (vemurafenib and dabrafenib) have initial encouraging effects in the treatment of melanoma but acquired resistance appears almost invariably after some months. Studies revealed mutually exclusive NRAS and BRAF activating mutations driving the MAPK/ERK pathway among human melanomas. Although combination therapy exerts significantly better antitumor cell efficacy, complete remission is rarely achieved. To employ an alternative approach, we have targeted the Hedgehog/GLI pathway, which is deregulated in melanomas, through the GLI1/2 inhibitor GANT61, alone or accompanied with the treatment by the BCL2 family inhibitor obatoclax in 9 melanoma cell lines. Thus, we targeted melanoma cells irrespective of their NRAS or BRAF mutational status. After GANT61 treatment, the cell viability was drastically diminished via apoptosis, as substantial nuclear DNA fragmentation was detected. In all tested melanoma cell lines, the combined treatment was more efficient than the application of each drug alone at the end of the cell growth with inhibitors. GANT61 was efficient also alone in most cell lines without the addition of obatoclax, which had only a limited effect when used as a single drug. In most cell lines, tumor cells were eradicated after 5-9 days of combined treatment in colony outgrowth assay. To conclude, GANT61 treatment might become a hopeful and effective anti-melanoma targeted therapy, especially when combined with the BCL2 family inhibitor obatoclax.

• MeSH
• antitumorózní látky farmakologie   MeSH
• GTP-fosfohydrolasy genetika   MeSH
• indoly MeSH
• jaderné proteiny antagonisté a inhibitory   metabolismus   MeSH
• lidé MeSH
• melanom farmakoterapie   genetika   metabolismus   MeSH
• membránové proteiny genetika   MeSH
• mutace MeSH
• nádorové buněčné linie MeSH
• proliferace buněk účinky léků   MeSH
• protein Gli2 s motivem zinkových prstů MeSH
• protoonkogenní proteiny B-Raf genetika   MeSH
• pyridiny farmakologie   MeSH
• pyrimidiny farmakologie   MeSH
• pyrroly farmakologie   MeSH
• synergismus léků MeSH
• transkripční faktory Krüppel-like antagonisté a inhibitory   metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antitumorózní látky MeSH
• BRAF protein, human MeSH Prohlížeč
• GANT 61 MeSH Prohlížeč
• GLI2 protein, human MeSH Prohlížeč
• GTP-fosfohydrolasy MeSH
• indoly MeSH
• jaderné proteiny MeSH
• membránové proteiny MeSH
• NRAS protein, human MeSH Prohlížeč
• obatoclax MeSH Prohlížeč
• protein Gli2 s motivem zinkových prstů MeSH
• protoonkogenní proteiny B-Raf MeSH
• pyridiny MeSH
• pyrimidiny MeSH
• pyrroly MeSH
• transkripční faktory Krüppel-like MeSH

BACKGROUND: Pituitary development and GH secretion are orchestrated by multiple genes including GH1, GHRHR, GLI2, HESX1, LHX3, LHX4, PROP1, POU1F1, and SOX3. We aimed to assess their mutation frequency and clinical relevance in children with severe GH deficiency (GHD). METHODS: The Genetics and Neuroendocrinology of Short Stature International Study (GeNeSIS; Clinical Trial Registry Number: NCT01088412) was a prospective, open-label, observational research program for pediatric patients receiving GH treatment, conducted in 30 countries between 1999 and 2015. The study included a sub-study to investigate mutations in the genes listed above. PCR products from genomic blood cell DNA were analyzed by Sanger sequencing. DNA variants were classified as pathogenic according to the recommendations of the American College of Medical Genetics and Genomics. Demographic, auxologic, and endocrine data at baseline and during GH treatment were documented and related to the genotyping results. FINDINGS: The analysis comprised 917 patients. In 92 patients (10%) 33 mutations were found, 16 previously described and 17 novel (52%). Mutation carriers were significantly younger, shorter, and more slowly growing than non-carriers. In general, their peak values in GH stimulation tests were very low; however, in 15/77 (20%) patients with GH1, PROP1, and SOX3 mutations they were only moderately diminished (3-6 μg/L). Two patients with a GH1 mutation developed TSH deficiency and one ADH deficiency. Using logistic multi-regression analysis, significant indicators of a mutation were combined pituitary hormone deficiency, greater patient-parent height difference (SDS), low GH peak, and young age. Final height SDS gain in mutation carriers (mean ± SD 3.4 ± 1.4) was greater than in non-carriers (2.0 ± 1.4; P < .001) and in patients with non-GHD short stature. INTERPRETATION: DNA testing for mutations in children with severe GHD shows a positive finding in approximately 10%. Phenotypes of mutation carriers can be variable. The benefit for clinical practice justifies DNA testing as an important component in the diagnostic work-up of patients with severe GHD. FUND: Eli Lilly and Company, Indianapolis, IN, USA. ClinicalTrials.com registration: NCT01088412.

• Klíčová slova
• Genetics, Growth hormone deficiency, Hypopituitarism, Pituitary, Short stature,
• MeSH
• dítě MeSH
• fenotyp MeSH
• homeodoménové proteiny genetika   MeSH
• hypofýza růst a vývoj   metabolismus   MeSH
• hypofyzární nanismus genetika   metabolismus   patofyziologie   MeSH
• jaderné proteiny genetika   MeSH
• lidé MeSH
• lidský růstový hormon genetika   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mutace * MeSH
• mutační analýza DNA MeSH
• předškolní dítě MeSH
• prospektivní studie MeSH
• protein Gli2 s motivem zinkových prstů genetika   MeSH
• proteiny s homeodoménou LIM genetika   MeSH
• receptory hormonů regulujících hypofyzární hormony MeSH
• receptory neuropeptidů MeSH
• transkripční faktor Pit-1 genetika   MeSH
• transkripční faktory SOXB1 genetika   MeSH
• transkripční faktory genetika   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• pozorovací studie MeSH
• Názvy látek
• GHRHR protein, human MeSH Prohlížeč
• GLI2 protein, human MeSH Prohlížeč
• HESX1 protein, human MeSH Prohlížeč
• homeodoménové proteiny MeSH
• jaderné proteiny MeSH
• Lhx3 protein MeSH Prohlížeč
• LHX4 protein, human MeSH Prohlížeč
• lidský růstový hormon MeSH
• POU1F1 protein, human MeSH Prohlížeč
• Prophet of Pit-1 protein MeSH Prohlížeč
• protein Gli2 s motivem zinkových prstů MeSH
• proteiny s homeodoménou LIM MeSH
• receptory hormonů regulujících hypofyzární hormony MeSH
• receptory neuropeptidů MeSH
• SOX3 protein, human MeSH Prohlížeč
• transkripční faktor Pit-1 MeSH
• transkripční faktory SOXB1 MeSH
• transkripční faktory MeSH

PURPOSE: Medulloblastoma comprises four distinct molecular subgroups: WNT, SHH, Group 3, and Group 4. Current medulloblastoma protocols stratify patients based on clinical features: patient age, metastatic stage, extent of resection, and histologic variant. Stark prognostic and genetic differences among the four subgroups suggest that subgroup-specific molecular biomarkers could improve patient prognostication. PATIENTS AND METHODS: Molecular biomarkers were identified from a discovery set of 673 medulloblastomas from 43 cities around the world. Combined risk stratification models were designed based on clinical and cytogenetic biomarkers identified by multivariable Cox proportional hazards analyses. Identified biomarkers were tested using fluorescent in situ hybridization (FISH) on a nonoverlapping medulloblastoma tissue microarray (n = 453), with subsequent validation of the risk stratification models. RESULTS: Subgroup information improves the predictive accuracy of a multivariable survival model compared with clinical biomarkers alone. Most previously published cytogenetic biomarkers are only prognostic within a single medulloblastoma subgroup. Profiling six FISH biomarkers (GLI2, MYC, chromosome 11 [chr11], chr14, 17p, and 17q) on formalin-fixed paraffin-embedded tissues, we can reliably and reproducibly identify very low-risk and very high-risk patients within SHH, Group 3, and Group 4 medulloblastomas. CONCLUSION: Combining subgroup and cytogenetic biomarkers with established clinical biomarkers substantially improves patient prognostication, even in the context of heterogeneous clinical therapies. The prognostic significance of most molecular biomarkers is restricted to a specific subgroup. We have identified a small panel of cytogenetic biomarkers that reliably identifies very high-risk and very low-risk groups of patients, making it an excellent tool for selecting patients for therapy intensification and therapy de-escalation in future clinical trials.

• MeSH
• čipová analýza tkání MeSH
• cytogenetika MeSH
• dítě MeSH
• hodnocení rizik MeSH
• hybridizace in situ fluorescenční MeSH
• jaderné proteiny genetika   MeSH
• kojenec MeSH
• lidé MeSH
• lidské chromozomy, pár 11 MeSH
• lidské chromozomy, pár 14 MeSH
• meduloblastom genetika   mortalita   patologie   terapie   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• nádorové biomarkery genetika   MeSH
• prediktivní hodnota testů MeSH
• předškolní dítě MeSH
• prognóza MeSH
• proporcionální rizikové modely MeSH
• protein Gli2 s motivem zinkových prstů MeSH
• proteiny hedgehog * genetika   MeSH
• proteiny Wnt * genetika   MeSH
• protoonkogenní proteiny c-myc genetika   MeSH
• regulace genové exprese u nádorů MeSH
• reprodukovatelnost výsledků MeSH
• rizikové faktory MeSH
• stanovení celkové genové exprese MeSH
• transkripční faktory Krüppel-like genetika   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• GLI2 protein, human MeSH Prohlížeč
• jaderné proteiny MeSH
• MYC protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• protein Gli2 s motivem zinkových prstů MeSH
• proteiny hedgehog * MeSH
• proteiny Wnt * MeSH
• protoonkogenní proteiny c-myc MeSH
• SHH protein, human MeSH Prohlížeč
• transkripční faktory Krüppel-like MeSH

Ptch receptors 1 and 2 mediate Hedgehog signaling pivotal for organ development and homeostasis. In contrast to embryonic lethal Ptch1 -/- phenotype, Ptch2 -/- mice display no effect on gross phenotype. In this brief report, we provide evidence of changes in the putative incisor mesenchymal stem cell (MSC) niches that contribute to accelerated incisor growth, as well as intriguing changes in the bones and skin which suggest a role for Ptch2 in the regulation of MSCs and their regenerative potential. We employed histological, immunostaining, and computed tomography (µCT) analyses to analyze morphological differences between Ptch2 -/- and wild-type incisors, long bones, and skins. In vitro CFU and differentiation assays were used to demonstrate the MSC content and differentiation potential of Ptch2 -/- bone marrow stromal cells. Wound healing assay was performed in vivo and in vitro on 8-week-old mice to assess the effect of Ptch2 on the wound closure. Loss of Ptch2 causes increases in the number of putative MSCs in the continuously growing incisor, associated with increased vascularization observed in the tooth mesenchyme and the neurovascular bundle. Increased length and volume of Ptch2 -/- bones is linked with the increased number and augmented in vitro differentiation potential of MSCs in the bone marrow. Dynamic changes in the Ptch2 -/- skin thickness relate to changes in the mesenchymal compartment and impact the wound closure potential. The effects of Ptch2 abrogation on the postnatal MSCs suggest a crucial role for Ptch2 in Hedgehog signaling regulation of the organ regenerative potential.

• Klíčová slova
• Gli1, Gli2, Hedgehog pathway, Ptch1, Ptch2, stem cells,
• Publikační typ
• časopisecké články MeSH

Survivin, an important antiapoptotic protein, is expressed in tumors, whereas in normal tissues the expression of this protein is extremely low, defining a role for survivin as a cancer gene. Survivin exhibits multifunctional activity in tumor cells. However, why survivin expression is sharply and invariably restricted to tumor tissue remains unclear. Here, we identified 11 putative consensus binding sites for GLI transcription factors in the survivin promoter and characterized the promoter activity. Inhibitors of the Hedgehog/GLI pathway, cyclopamine and GANT61, decreased the promoter activity in reporter assays. ΔNGLI2 (which lacks the repressor domain) was the most potent vector in activating the survivin promoter-reporter. Moreover, GANT61, a GLI1/2 inhibitor, repressed endogenous survivin protein and mRNA expression in most cells across a large panel of tumor cell lines. Chromatin immunoprecipitation showed GLI2 binding to the survivin promoter. The ectopic GLI2-evoked expression of endogenous survivin was observed in normal human fibroblasts. GANT61 decreased survivin level in nude mice tumors, mimicking the activity of GANT61 in cultured cells. The immunohistochemistry and double immunofluorescence of human tumors revealed a correlation between the tissue regions showing high GLI2 and survivin positivity. Thus, these results demonstrated that survivin is a classical transcriptional target of GLI2, a Hedgehog pathway signaling effector. This potentially reflects the high expression of survivin in human tumor cells. As the Hedgehog pathway is upregulated in virtually all types of cancer cells, these findings substantially contribute to the explanation of uniform survivin expression in tumors as a potential target for the development of a more effective treatment of cancers through the inhibition of GLI2 to restrain survivin activity.

• MeSH
• inhibitory apoptózy genetika   metabolismus   MeSH
• lidé MeSH
• myši nahé MeSH
• myši MeSH
• proteiny hedgehog metabolismus   MeSH
• represorové proteiny genetika   metabolismus   MeSH
• signální transdukce MeSH
• survivin MeSH
• transfekce MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Birc5 protein, mouse MeSH Prohlížeč
• inhibitory apoptózy MeSH
• proteiny hedgehog MeSH
• represorové proteiny MeSH
• survivin MeSH

Sonic hedgehog medulloblastoma encompasses a clinically and molecularly diverse group of cancers of the developing central nervous system. Here, we use unbiased sequencing of the transcriptome across a large cohort of 250 tumors to reveal differences among molecular subtypes of the disease, and demonstrate the previously unappreciated importance of non-coding RNA transcripts. We identify alterations within the cAMP dependent pathway (GNAS, PRKAR1A) which converge on GLI2 activity and show that 18% of tumors have a genetic event that directly targets the abundance and/or stability of MYCN. Furthermore, we discover an extensive network of fusions in focally amplified regions encompassing GLI2, and several loss-of-function fusions in tumor suppressor genes PTCH1, SUFU and NCOR1. Molecular convergence on a subset of genes by nucleotide variants, copy number aberrations, and gene fusions highlight the key roles of specific pathways in the pathogenesis of Sonic hedgehog medulloblastoma and open up opportunities for therapeutic intervention.

• MeSH
• dítě MeSH
• dospělí MeSH
• genetická variace MeSH
• genové regulační sítě MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• meduloblastom genetika   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• nádory mozečku genetika   MeSH
• předškolní dítě MeSH
• proteiny hedgehog genetika   MeSH
• regulace genové exprese u nádorů * MeSH
• signální transdukce genetika   MeSH
• transkriptom * MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• proteiny hedgehog MeSH
• SHH protein, human MeSH Prohlížeč

In melanoma and other cancers, invasion, epithelial-to-mesenchymal transition, metastasis and cancer stem cell maintenance are regulated by transcription factors including the Snail family. Slug (Snail2) protein generally supports migration and apoptosis resistance. However, its role in melanoma is not completely understood. The present study investigated the transcriptional regulation of the SLUG gene in melanoma. It demonstrated that SLUG is under the control of the Hedgehog/GLI signaling pathway and is activated predominantly by the transcription factor GLI2. The SLUG gene promoter contains a high number of GLI-binding sites. Slug expression is activated by GLI factors in reporter assays and inhibited by GANT61 (GLI inhibitor) and cyclopamine (SMO inhibitor). SLUG mRNA levels are lowered by GANT61 as assessed by reverse transcription-quantitative PCR. Chromatin immunoprecipitation revealed abundant binding of factors GLI1-3 in the four subregions of the proximal SLUG promoter. Notably, melanoma-associated transcription factor (MITF) is an imperfect activator of the SLUG promoter in reporter assays, and downregulation of MITF had no effect on endogenous Slug protein levels. Immunohistochemical analysis confirmed the above findings and showed MITF-negative regions in metastatic melanoma that were positive for GLI2 and Slug. Taken together, the results demonstrated a previously unrecognized transcriptional activation mechanism of the SLUG gene, which may represent its main regulation of expression in melanoma cells.

• Klíčová slova
• GLI family zinc finger, Hedgehog signaling, Slug, melanoma, melanoma-associated transcription factor,
• MeSH
• apoptóza MeSH
• lidé MeSH
• melanom * genetika   MeSH
• proteiny hedgehog * genetika   MeSH
• signální transdukce MeSH
• transkripční faktory genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• proteiny hedgehog * MeSH
• transkripční faktory MeSH