Dataset Information

Oncogenic mutations in the Gα-protein GNA11 convey a shorter disease specific survival and are more strongly associated with loss of BAP1 and chromosomal alterations than GNAQ mutations in uveal melanoma

ABSTRACT: 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.

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Martina Bartolucci

LAB HEAD: Andrea Petretto

PROVIDER: PXD030217 | Pride | 2022-06-22

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	GNAQ_GNA11.zip	Other
	HomoSapien_tax9606_UP000005640.fasta	Fasta
	Pfeffer_F_11_Mut_031.raw	Raw
	Pfeffer_F_11_Mut_2_032.raw	Raw
	Pfeffer_F_11_Mut_3_033.raw	Raw

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In uveal melanoma Gα-protein GNA11 mutations convey a shorter disease-specific survival and are more strongly associated with loss of BAP1 and chromosomal alterations than Gα-protein GNAQ mutations.

Piaggio Francesca F Croce Michela M Reggiani Francesco F Monti Paola P Bernardi Cinzia C Ambrosio Marianna M Banelli Barbara B Dogrusöz Mehmet M Jockers Ralf R Bordo Domenico D Puzone Roberto R Viaggi Silvia S Coviello Domenico D Lanza Francesco B FB Bartolucci Martina M Petretto Andrea A Mosci Carlo C Gangemi Rosaria R van der Velden Pieter A PA Jager Martine J MJ Pfeffer Ulrich U Amaro Adriana A

European journal of cancer (Oxford, England : 1990) 20220514

<h4>Background and aim of the study</h4>Mutations in the Gα-genes GNAQ and GNA11 are found in 85-90% of uveal melanomas (UM). Aim of the study is to understand whether the mutations in both genes differentially affect tumor characteristics and outcome and if so, to identify potential mechanisms.<h4>Methods</h4>We analyzed the association between GNAQ and GNA11 mutations with disease-specific survival, gene expression profiles, and cytogenetic alterations in 219 UMs. We used tandem-affinity-purif ...[more]

PMID: 35580369

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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

Project description:Uveal melanoma (UM) is the most prevalent cancer of the eye in adults, with a highly aggressive form of metastasis that is refractory to current therapies. UM is driven by aberrant activation of the Gαq pathway by hotspot activating mutation of GNAQ/GNA11, with few additional genetic aberrations. Despite this, there are limited effective targeted therapies currently available against the treatment of UM and mUM. Here, we performed a high-throughput chemogenetic drug screen in GNAQ-mutant UM contrasted with that of BRAF-mutant skin cutaneous melanoma, as a network chemical biology-based approach to identify therapeutic agents that target the mechanistic underpinnings driving UM. We observed strong genotype-driven drug sensitivities, and identified several drug classes with preferential activity against UM using a method termed Drug Set Enrichment Analysis (DSEA). Among them, we found an enrichment for PKC inhibitors, and identified one compound LXS-196, with the highest preferential activity against UM. Our investigation into the mechanism of action of LXS-196 revealed that in addition to inhibiting the Gq-ERK pathway, unlike other PKC inhibitors, this drug also reduced FAK activity, a recently identified mediator of non-canonical Gαq-driven oncogenic signaling. Kinome profiling revealed that LXS-196 acts as a multi-targeted kinase inhibitor, with high preference for PKC as well as PKN/PRK, the latter a poorly investigated AGC kinase that is activated directly by RhoA. This primes LXS-196 to target cell-essential pathways that drive tumor growth in UM by targeting both PKC, in addition to FAK. Moreover, we find that PKN is activated by GNAQ downstream from RhoA, thereby contributing to FAK stimulation. These findings expose a signaling vulnerability that can be targeted pharmacologically. Ultimately, dual PKC and PKN inhibition by LXS-196 acts synergistically with FAK inhibitors (FAKi) to halt UM growth and promote cytotoxic cell death in vitro and in preclinical metastatic mouse models, thus providing a highly translatable therapeutic multimodal precision strategy against mUM.