RNA-seq analysis of MEK or ERK1/2 inhibitor treatment in uveal melanoma cell lines.
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE228090 | GEO | 2023/12/20
REPOSITORIES: GEO
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Project description:Mutations of the Gα-proteins GNAQ and GNA11 are found in 85-90% of uveal melanomas (UMs). We analyzed the association of GNAQ and GNA11 mutations with disease-specific survival, gene expression profiles, and cytogenetic alterations in 219 UMs. Our analysis showed a shorter disease-specific survival of GNA11 mutated cases as compared to those carrying a GNAQ mutation (median months: 26.97 [95% CI 25.02-37.08], vs not reached, p=0.058; HR=1.97 [95%CI 1.12-3.46], p=0.02). The GNA11 mutation was associated with expression alterations of the BRCA1-associated protein 1 (BAP1; p=0.0005), chr3 monosomy (p=0.0002), chr8q amplification (p=0.038), the combination of the latter two (p=0.0002), and, inversely, with chr6p amplification (p=0.003). We used tandem-affinity-purification and mass spectrometry to show that the two G-proteins have different protein interaction partners. The Tet Methylcytosine Dioxygenase 2, TET2, a protein that is involved in DNA-demethylation, physically interacts with GNAQ but not with GNA11 as confirmed by immunoprecipitation analyses. High risk UM shows a clearly different DNA-methylation pattern and different control of DNA-methylation by the two G-proteins might, at least in part, explain the different association with UM progression.
2022-06-22 | PXD030217 | Pride
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
Project description:Mutations in the Gα-genes GNAQ and GNA11 are found in 85-90% of uveal melanomas (UMs). We analyzed the association between GNAQ and GNA11 mutations with disease-specific survival, gene expression profiles, and cytogenetic alterations in 219 UMs. Our analysis showed a shorter disease-specific survival of GNA11-mutated cases as compared to those carrying a GNAQ mutation (HR=1.97 [95%CI 1.12-3.46], p=0.02). GNA11 mutated UMs displayed a shorter, but not significant, median months survival of 26.97 [95% CI 25.02-37.08] versus GNAQ patients median months survival of 31.5 [95% CI 30.97-53.51]. The GNA11 mutation was also associated with: i) an increased frequency of loss of BRCA1-associated protein 1 (BAP1) expression (p=0.0005), ii) monosomy of chromosome 3 (p<0.001), iii) amplification of chr8q (p=0.038), iv) the combination of the latter two (p=0.0002), and inversely with v) chr6p gain (p=0.003). We used tandem-affinity-purification and mass spectrometry to show that the two derived G-proteins have different protein interaction partners. Specifically, the Tet Methylcytosine Dioxygenase 2(TET2), a protein that is involved in DNA demethylation, physically interacts with the GNAQ protein but not with GNA11, as confirmed by immunoprecipitation analyses. High-risk UM cases show a clearly different DNA-methylation pattern, suggesting that a different regulation of DNA methylation by the two G-proteins might convey a different risk of progression.
2023-02-27 | GSE197656 | 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:Uveal melanoma (UM) is the most common intraocular tumor in adults, and its pathogenesis is only partly understood. UM can develop de novo from normal choroidal melanocytes (NCMs) or from pre-existing nevi that stem from NCMs and are thought to harbour UM-initiating mutations, most commonly in GNAQ or GNA11. However, there is a complete lack of commercially available NCM cell lines and no detailed protocol for developing oncogene-mutated CM line (MutCM) to study UM development. The purpose of this study was to establish and characterize premalignant CM models from human donor eyes to recapitulate the cell populations at the origin of UM.
2024-11-25 | GSE273527 | GEO
Project description:Uveal melanoma (UM) is the most common intraocular tumor in adults, and its pathogenesis is only partly understood. UM can develop de novo from normal choroidal melanocytes (NCMs) or from pre-existing nevi that stem from NCMs and are thought to harbour UM-initiating mutations, most commonly in GNAQ or GNA11. However, there is a complete lack of commercially available NCM cell lines and no detailed protocol for developing oncogene-mutated CM line (MutCM) to study UM development. The purpose of this study was to establish and characterize premalignant CM models from human donor eyes to recapitulate the cell populations at the origin of UM.
2024-11-25 | PXD047179 | Pride
Project description:Background and Aims: Uveal melanoma (UM), the most common primary intraocular tumor in adults, is characterized by marked variability in its ability to metastasize. In fact, up to half the patients with UM develop distant metastases, most commonly to the liver. Mutations in alpha G-protein subunits, GNAQ and GNA11, are found in most primary UM, in a mutually exclusive pattern. Also, mutations in SF3B1, EIF1AX, and BAP1, are associated with markedly distinct prognoses Targeted therapy with MAP kinase inhibitors has been unsuccessful in the treatment of uveal melanoma (UM). The constitutive activation of GNAq and GNA11, typical of MU, determines the activation not only of the MAP kinases but also of the transcription factors YAP / TAZ. It is known that when statins are used to inhibit the HMGCR activity in the mevalonate pathway, the nuclear localization and transcriptional activity of YAP-TAZ are also inhibited mediating a potential anti-cancer activity Methods: We investigated the sensitivity to drugs of a panel of 6 lines of MU, two metastatic and 4 derived from primary tumors, of the latter two were monosomic. We established the IC50s of the individual drugs and the effects of the combinations. The synergistic effects of drugs were studied using MTT proliferation tests, cell cycle and apoptosis. The most synergistic combination was tested in vivo in NSG mice. Results: The synergistic concentrations of trametinib and cerivastatin (T+C) induced a massive arrest of proliferation and cell cycle and an increase of apoptosis 72 hours after the start of treatment particularly in the monosomic, BAP1 mutated UPMM3 cell line. The synergistic effect of T+C was reached after 7 instead of 3 days. In the metastatic OMM2.5 cell line. T+C treatments reduced ERK and AKT phosphorylation and increased the cytoplasmatic, inactive form of YAP. T+C treatment of NSG mice transplanted with the monosomic UM cell line reduced significantly tumor growth. Conclusion: This study suggests that statins potentiate the efficacy of MEK inhibitors in the therapy of UM
2022-08-31 | GSE212219 | 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. Using a melanocytic lineage expression system, we showed that patient-derived mutations of GNAQ (GNAQQ209L) or its upstream CYSLTR2 receptor (CYSLTR2L129Q) both drive UM when combined with a cooperating mutation, tp53M214K/M214K. The tumor-initiating potential of the major GNAQ/11 effector pathways, YAP and PLCβ-ERK, was also investigated in this system, and thus showed that while activated YAP (YAPAA) induced UM with high potency, the patient-derived PLC4 mutation (PLCB4D630Y) very rarely yielded UM tumors in the tp53M214K/M214K context. Remarkably, mitfa deficiency was profoundly UM promoting, dramatically accelerating the onset and progression of tumors induced by Tg(GNAQQ209L);tp53M214K/M214K or Tg(CYSLTR2L129Q);tp53M214K/M214K. Moreover, mitfa loss was sufficient to cooperate with GNAQQ209L to drive tp53-wildtype UM development, and allowed Tg(PLCB4D630Y);tp53M214K/M214K melanocyte lineage cells to readily form tumors. Notably, all of the mitfa-/- UM tumors, including those arising in Tg(mitfa:PLCB4D630Y);tp53M214K/M214K;mitfa-/- fish, displayed nuclear YAP while lacking hyperactive ERK indicative of PLCβ signaling. Collectively, these data show that YAP signaling is the major mediator of UM, and that MITF acts as bona fide tumor suppressor in UM, in direct opposition to its essential role in CM.
2022-04-27 | GSE190802 | GEO
Project description:We developed an isogenic melanocytic cellular system and systematically examined the hotspot mutations in GNAQ (e.g., G48V, R183Q, Q209L) and CYSLTR2 (e.g. L129Q) in human uveal melanoma. Biochemical and cell viability assays validated YM-254890 as a potent inhibitor of cell signaling and growth. Human uveal melanoma cells and mouse models recapitulated this finding, indicating that YM is also effective in vivo. Combination of YM and MEK inhibition leads to further decreases in MAPK pathway gene expression and cell viability. Furthermore, our cellular and mouse models show that combination leads to long term signaling inhibition and significant decreases in tumor burden.
2020-12-31 | GSE160112 | GEO
Project description:We developed an isogenic melanocytic cellular system and systematically examined the hotspot mutations in GNAQ (e.g., G48V, R183Q, Q209L) and CYSLTR2 (e.g. L129Q) in human uveal melanoma. Biochemical and cell viability assays validated YM-254890 as a potent inhibitor of cell signaling and growth. Human uveal melanoma cells and mouse models recapitulated this finding, indicating that YM is also effective in vivo. Combination of YM and MEK inhibition leads to further decreases in MAPK pathway gene expression and cell viability. Furthermore, our cellular and mouse models show that combination leads to long term signaling inhibition and significant decreases in tumor burden.
2020-12-31 | GSE152705 | GEO