Ultrasensitive profiling of UV mutations in facial tumors in TSC
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PROVIDER: PRJNA836690 | ENA |
REPOSITORIES: ENA
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Project description:Ultrasensitive profiling of UV mutations in facial tumors in TSC
| PRJNA836691 | ENA
Project description:BackgroundTuberous sclerosis complex (TSC) is a neurogenetic syndrome due to loss-of-function mutations in TSC2 or TSC1, characterized by tumors at multiple body sites, including facial angiofibroma (FAF). Here, an ultrasensitive assessment of the extent and range of UV-induced mutations in TSC facial skin was performed.MethodsA multiplex high-sensitivity PCR assay (MHPA) was developed, enabling mutation detection at extremely low (<0.1%) variant allele frequencies (VAFs).ResultsMHPA assays were developed for both TSC2 and TP53, and applied to 81 samples, including 66 skin biopsies. UV-induced second-hit mutation causing inactivation of TSC2 was pervasive in TSC facial skin with an average of 4.8 mutations per 2-mm biopsy at median VAF 0.08%, generating more than 150,000 incipient facial tumors (subclinical "micro-FAFs") in the average TSC subject. The MHPA analysis also led to the identification of a refined UV-related indel signature and a recurrent complex mutation pattern, consisting of both a single-nucleotide or dinucleotide variant and a 1- to 9-nucleotide deletion, in cis.ConclusionTSC facial skin can be viewed as harboring a patchwork of clonal fibroblast proliferations (micro-FAFs) with indolent growth, a small proportion of which develop into clinically observable FAF. Our observations also expand the spectrum of UV-related mutation signatures.FundingThis work was supported by the TSC Alliance; the Engles Family Fund for Research in TSC and LAM; and the NIH, National Heart, Lung, and Blood Institute (U01HL131022-04 and Intramural Research Program).
| S-EPMC9106361 | biostudies-literature
Project description:We report the global gene expression effects in TSC-deficient and TSC wild type cells after treatemnet with. CDK7 inhibitor(THZ1), we performed RNA-Seq on TSC1-null HCV.29 cells and TSC1-intact HCV.29 cells treated with 30nM THZ1, and with 100nM THZ1, for 6 hours. Many genes showed marked changes in expression in comparison with control untreated cells. Furthermore, 1128 genes showed > 5-fold lower expression in 30nM THZ1-treated TSC1-null HCV.29 cells compared with 30nM THZ1-treated TSC1-intact HCV.29 cells.
2019-08-08 | GSE135527 | GEO
Project description:Patients with tuberous sclerosis complex (TSC) develop hamartomas containing biallelic inactivating mutations in either TSC1 or TSC2, resulting in mammalian target of rapamycin (mTOR) activation. Hamartomas overgrow epithelial and mesenchymal cells in TSC skin. The pathogenetic mechanisms for these changes had not been investigated, and the existence or location of cells with biallelic mutations (âtwo-hitâ cells) that resulted in mTOR activation was unclear. We compared TSC skin hamartomas (facial angiofibromas and periungual fibromas) to normal-appearing skin of the same patient, and observed more proliferation and mTOR activation in hamartoma epidermis. âTwo-hitâ cells were not detected in the epidermis. Fibroblast-like cells in the dermis, however, exhibited allelic deletion of TSC2, in both touch preparations of fresh tumor samples and cells grown from TSC skin tumors, suggesting that increased epidermal proliferation and mTOR activation were not caused by second-hit mutations in the keratinocytes but by mesenchymal-epithelial interactions. Gene expression arrays, used to identify potential paracrine factors released by mesenchymal cells, revealed more epiregulin mRNA in fibroblast-like angiofibroma and periungual fibroma cells than in fibroblasts from normal-appearing skin of the same patient. Elevation of epiregulin mRNA was confirmed using real-time PCR, and increased amounts of epiregulin protein were demonstrated using immunoprecipitation and ELISA. Epiregulin stimulated keratinocyte proliferation and phosphorylation of ribosomal protein S6 in vitro. These results suggest that hamartomatous TSC skin tumors are induced by paracrine factors released by âtwo-hitâ cells in the dermis, and that proliferation with mTOR activation of the overlying epidermis is an effect of epiregulin. Experiment Overall Design: The study is of case/control design with biological replication. Tumor (case) and normal (control) fibroblast cells were isolated from each of four patients (biological replicates).
2008-04-06 | E-GEOD-9715 | biostudies-arrayexpress
Project description:Patients with tuberous sclerosis complex (TSC) develop hamartomas containing biallelic inactivating mutations in either TSC1 or TSC2, resulting in mammalian target of rapamycin (mTOR) activation. Hamartomas overgrow epithelial and mesenchymal cells in TSC skin. The pathogenetic mechanisms for these changes had not been investigated, and the existence or location of cells with biallelic mutations (“two-hit” cells) that resulted in mTOR activation was unclear. We compared TSC skin hamartomas (facial angiofibromas and periungual fibromas) to normal-appearing skin of the same patient, and observed more proliferation and mTOR activation in hamartoma epidermis. “Two-hit” cells were not detected in the epidermis. Fibroblast-like cells in the dermis, however, exhibited allelic deletion of TSC2, in both touch preparations of fresh tumor samples and cells grown from TSC skin tumors, suggesting that increased epidermal proliferation and mTOR activation were not caused by second-hit mutations in the keratinocytes but by mesenchymal-epithelial interactions. Gene expression arrays, used to identify potential paracrine factors released by mesenchymal cells, revealed more epiregulin mRNA in fibroblast-like angiofibroma and periungual fibroma cells than in fibroblasts from normal-appearing skin of the same patient. Elevation of epiregulin mRNA was confirmed using real-time PCR, and increased amounts of epiregulin protein were demonstrated using immunoprecipitation and ELISA. Epiregulin stimulated keratinocyte proliferation and phosphorylation of ribosomal protein S6 in vitro. These results suggest that hamartomatous TSC skin tumors are induced by paracrine factors released by “two-hit” cells in the dermis, and that proliferation with mTOR activation of the overlying epidermis is an effect of epiregulin. Keywords: Disease state analysis
2008-03-01 | GSE9715 | GEO
Project description:EOMES is an essential transcription factor (TF) for murine trophoblast stem cell (TSC) maintenance. Despite that, the details of EOMES' function at the molecular level remain obscure. Here, we carried out rapid immunoprecipitation and mass spectrometry of endogenous protein (RIME) to identify TSC-specific protein interactors on EOMES. In addition to other established TFs involved in TSC maintenance, we found that EOMES interacts with several chromatin remodeller subunits, including BRG1. By exploiting an Eomes-degron system, we acutely depleted EOMES protein in TSCs and in parallel inhibited BRG1 function with small molecule BRM014. EOMES depletion and BRG1 inhibition resulted in reduced accessibility at largely overlapping genomic regions associated with TSC-related loci. Additionally, EOMES depletion results in misregulation of genes essential for TSC maintenance and function, as well as genes encoding cytoskeletal, cell-cell interaction and matricellular components.
2025-05-19 | GSE276261 | GEO
Project description:EOMES is an essential transcription factor (TF) for murine trophoblast stem cell (TSC) maintenance. Despite that, the details of EOMES' function at the molecular level remain obscure. Here, we carried out rapid immunoprecipitation and mass spectrometry of endogenous protein (RIME) to identify TSC-specific protein interactors on EOMES. In addition to other established TFs involved in TSC maintenance, we found that EOMES interacts with several chromatin remodeller subunits, including BRG1. By exploiting an Eomes-degron system, we acutely depleted EOMES protein in TSCs and in parallel inhibited BRG1 function with small molecule BRM014. EOMES depletion and BRG1 inhibition resulted in reduced accessibility at largely overlapping genomic regions associated with TSC-related loci. Additionally, EOMES depletion results in misregulation of genes essential for TSC maintenance and function, as well as genes encoding cytoskeletal, cell-cell interaction and matricellular components.
2025-05-19 | GSE276042 | GEO
Project description:EOMES is an essential transcription factor (TF) for murine trophoblast stem cell (TSC) maintenance. Despite that, the details of EOMES' function at the molecular level remain obscure. Here, we carried out rapid immunoprecipitation and mass spectrometry of endogenous protein (RIME) to identify TSC-specific protein interactors on EOMES. In addition to other established TFs involved in TSC maintenance, we found that EOMES interacts with several chromatin remodeller subunits, including BRG1. By exploiting an Eomes-degron system, we acutely depleted EOMES protein in TSCs and in parallel inhibited BRG1 function with small molecule BRM014. EOMES depletion and BRG1 inhibition resulted in reduced accessibility at largely overlapping genomic regions associated with TSC-related loci. Additionally, EOMES depletion results in misregulation of genes essential for TSC maintenance and function, as well as genes encoding cytoskeletal, cell-cell interaction and matricellular components.
2025-05-19 | GSE276039 | GEO
Project description:Genomic Sites Ultrasensitive to Ultraviolet Radiation
| PRJNA564830 | ENA
Project description:Peg10 regulation of TSC differentiation
| PRJNA504005 | ENA