A novel somatic mutation in GNAQ in Sturge-Weber Syndrome provides insight into disease pathogenesis
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ABSTRACT:
PROVIDER: PRJNA822387 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR18568220_1.fastq.gz | Fastqsanger.gz | |||
| SRR18568220_2.fastq.gz | Fastqsanger.gz | |||
| SRR18568221_1.fastq.gz | Fastqsanger.gz | |||
| SRR18568221_2.fastq.gz | Fastqsanger.gz | |||
| SRR18568222_1.fastq.gz | Fastqsanger.gz |
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Project description:Sturge-Weber syndrome (SWS) is a sporadic, congenital, neuro-cutaneous disorder characterized by a mosaic, capillary malformation. SWS and isolated capillary malformations are caused by a somatic activating mutation in GNAQ encoding the G protein subunit alpha-q protein. The missense mutation R183Q is the sole GNAQ mutation identified thus far in affected tissues of 90% of SWS patients. In this study, we sequenced skin biopsies of affected capillary malformations from 9 patients. We identified the R183Q mutation in nearly all samples, but one sample exhibited a Q209R mutation. This new mutation occurs at the same residue as the constitutively-activating Q209L mutation, commonly seen in tumors. However, Q209R is a rare variant in this gene. To compare the effect of the Q209R mutation on downstream signaling, we performed reporter assays with a GNAQ-responsive reporter co-transfected with either GNAQ WT, R183Q, Q209L, Q209R, or C9X (representing a null allele). Q209L showed the highest reporter activation, with R183Q and Q209R showing significantly lower activation. To determine whether these mutations had similar or different downstream consequences we performed RNAseq analysis in microvascular endothelial cells (HMEC-1) electroporated with the same GNAQ variants. The R183 and Q209 missense variants caused extensive dysregulation of a broad range of transcripts compared to the WT or null allele, confirming that these are all activating mutations. However, the missense variants exhibited very few differentially expressed genes (DEGs) when compared to each other. These data suggest that these activating GNAQ mutations differ in magnitude of activation but have similar downstream effects.
2022-07-28 | GSE199978 | GEO
Project description:Sturge-Weber Syndrome (SWS) is a common vascular malformation caused by mutation of GNAQ in endothelial cells. No effective medical therapy exists for SWS, and therapy is mainly surgical. Progress in the medical treatment of SWS has been hindered by the lack of an animal model. We introduced mutant GNAQ into an immortalized murine endothelial cell line, MS1. The resultant cells form slow growing vascular malformations that morphologically and biochemically resemble human SWS. One of the genes upregulated as a result of mutant GNAQ is c-kit, which is a targetable event. Given that the FDA approved drug imatinib targets c-kit as well as its original target, bcr-abl, we evaluated the ability of imatinib to prevent the growth of vascular malformations in mice. We saw that imatinib significantly inhibits the growth of vascular malformations in vivo. Repurposing of imatinib for the treatment of SWS should be evaluated in a clinical trial.
2022-10-31 | GSE189023 | GEO
Project description:Autoantibody repertoire characterization provides insight into the pathogenesis of dissimilar autoimmune diseases
| PRJNA921887 | ENA
Project description:The Sturge-Weber syndrome is a sporadic congenital neurocutaneous disorder characterized by a port-wine stain affecting the skin in the distribution of the ophthalmic branch of the trigeminal nerve, abnormal capillary venous vessels in the leptomeninges of the brain and choroid, glaucoma, seizures, stroke, and intellectual disability. It has been hypothesized that somatic mosaic mutations disrupting vascular development cause both the Sturge-Weber syndrome and port-wine stains, and the severity and extent of presentation are determined by the developmental time point at which the mutations occurred. To date, no such mutation has been identified.We performed whole-genome sequencing of DNA from paired samples of visibly affected and normal tissue from 3 persons with the Sturge-Weber syndrome. We tested for the presence of a somatic mosaic mutation in 97 samples from 50 persons with the Sturge-Weber syndrome, a port-wine stain, or neither (controls), using amplicon sequencing and SNaPshot assays, and investigated the effects of the mutation on downstream signaling, using phosphorylation-specific antibodies for relevant effectors and a luciferase reporter assay.We identified a nonsynonymous single-nucleotide variant (c.548G?A, p.Arg183Gln) in GNAQ in samples of affected tissue from 88% of the participants (23 of 26) with the Sturge-Weber syndrome and from 92% of the participants (12 of 13) with apparently nonsyndromic port-wine stains, but not in any of the samples of affected tissue from 4 participants with an unrelated cerebrovascular malformation or in any of the samples from the 6 controls. The prevalence of the mutant allele in affected tissues ranged from 1.0 to 18.1%. Extracellular signal-regulated kinase activity was modestly increased during transgenic expression of mutant G?q.The Sturge-Weber syndrome and port-wine stains are caused by a somatic activating mutation in GNAQ. This finding confirms a long-standing hypothesis. (Funded by the National Institutes of Health and Hunter's Dream for a Cure Foundation.).
| S-EPMC3749068 | biostudies-literature
Project description:Local ancestry inference provides insight into Tilapia breeding programmes
| PRJEB38387 | ENA
Project description:Introduction: Capillary malformation (CM) occurs sporadically and is associated with Sturge-Weber syndrome (SWS). The somatic mosaic mutation in GNAQ (c.548GàA, p.R183Q) is enriched in endothelial cells (EC) in skin CM and SWS brain CM. Our goal was to investigate how the mutant G-protein a-q subunit (Gaq) alters EC signaling and disrupts capillary morphogenesis. Approach and results: We used lentiviral constructs to express p.R183Q or wild-type GNAQ in normal human endothelial colony forming cells (EC-R183Q and EC-WT respectively). EC-R183Q constitutively activated phospholipase-C β3 (PLCβ3), a downstream effector of Gαq. Activated PLCβ3 was also detected in human CM tissue sections. Bulk RNA-seq analyses of mutant versus wild-type EC indicated constitutive activation of protein kinase C (PKC), NF-κB and calcineurin signaling in EC-R183Q. Increased expression of downstream targets in these pathways, Angiopoetin-2 (ANGPT2), C-X-C Motif Chemokine Ligand 5 (CXCL5) and Down Syndrome Critical Region Protein 1.4 (DSCR1.4) were confirmed by qPCR and immunostaining of human CM tissue sections. The Gαq inhibitor YM-254890 reduced mRNA expression levels of these targets in EC-R183Q while the pan-PKC inhibitor AEB071 reduced ANGPT2 and CXCL5 but not DSCR1.4. EC-R183Q formed enlarged blood vessels in mice, reminiscent of those found in human CM, with robust endothelial ANGPT2. Conclusions: Gaq-R183Q, when expressed in ECs, establishes constitutively active PLCb3 signaling that leads to a pro-angiogenic, pro-inflammatory phenotype. EC-R183Q are sufficient to form CM-like vessels in mice which provides the first evidence that endothelial Gaq-R183Q is causative for CM.
2021-11-09 | GSE186998 | GEO
Project description:We used microarray to compare global gene expression profiles between 5 GNAQ/11 mutant uveal melanoma cell lines (GNAQ mutant: 92-1, omm1.3, mel270; GNA11 mutant: omm-gn11 and upmd-1) and 5 GNAQ/11 wild type melanoma cell lines(sk-mel-2, mm415, mm485, sk-mel-5 and mum2c). Uveal melanoma is the most common intraocular tumor that mainly metastasizes to the liver in about 50% patients. Over 80% of UMs harbor GNAQ or GNA11 activating mutation. Currently there is no effective treatment available for UM patients. Results provide insights into downstream signaling of oncogenic GNAQ/11 and identification of therapeutic targets in UM.
2017-07-01 | GSE93666 | GEO
Project description:Droplet digital PCR (ddPCR), a method for measuring target nucleic acid sequence quantity, is useful for determining somatic mutation rates using TaqMan probes. In this study, the detection limit of copy numbers of test DNA by ddPCR was determined based on Poisson distribution. Peptide nucleic acid (PNA), which strongly hybridises to target lesions, can inhibit target amplification by PCR. Therefore, by combination of PCR with PNA and ddPCR (PNA-ddPCR), the detection limit could be lowered. We reanalysed a somatic GNAQ mutation (c.548G > A) in patients with Sturge-Weber syndrome (SWS) using ddPCR and PNA-ddPCR. Importantly, among three patients previously found to be mutation negative by next-generation sequencing, two patients had the GNAQ mutation with a mutant allele frequency of less than 1%. Furthermore, we were able to find the same mutation in blood leukocyte or saliva DNA derived from four out of 40 SWS patients. Vascular anomalies and blood leukocytes originate from endothelial cells and haemangioblasts, respectively, which are both of mesodermal origin. Therefore, blood leukocytes may harbour the GNAQ mutation, depending on the time when the somatic mutation is acquired. These data suggest the possibility of diagnosis using blood DNA in some patients with SWS.
| S-EPMC4783707 | biostudies-literature
Project description:To investigate the transcriptional effects of GNAQ p.Q209L mutation on endothelial cells, bulk RNA-sequencing was performed on endothelial cells lines expressing GNAQ wildtype or mutant Q209L. Cells were also treated with Trametinib or vehicle control to elucidate the specific effects of MAPK/ERK signaling on these cells.
2022-10-25 | GSE216367 | GEO
Project description:Cutaneous melanoma (CM) and uveal melanoma (UM) both originate from the melanocytic lineage but are primarily driven by distinct oncogenic drivers, BRAF/NRAS or GNAQ/GNA11 respectively. The melanocytic master transcriptional regulator, MITF, is essential for both CM development and maintenance, but its role in UM is largely unexplored. Here, we use zebrafish models to dissect the key UM oncogenic signaling events, and establish the role of MITF in UM tumors. Remarkably, mitfa deficiency was profoundly UM promoting, dramatically accelerating the onset and progression of tumors induced by Tg(mitfa:GNAQQ209L);tp53M214K/M214K. To further explore the role of MITF in GNAQ-driven tumorigenesis, we performed phospho-proteomics and total proteomics on 5 zebrafish GNAQ Tg(mitfa:GNAQQ209L);tp53M214K/M214K tumors and 5 zebrafish GNAQ Tg(mitfa:GNAQQ209L);tp53M214K/M214K;mitfa-/- tumors.
2023-03-11 | PXD030527 | Pride