Search

Most uveal melanoma cases harbor activating mutations in either GNAQ or GNA11. Despite activation of the mitogen-activated protein kinase (MAPK) signaling pathway downstream of Gαq/11, there are no effective targeted kinase therapies for metastatic uveal melanoma. The human genome encodes numerous understudied kinases, also called the dark kinome. Identifying additional kinases regulated by Gαq/11 may uncover novel therapeutic targets for uveal melanoma. In this study, we treated GNAQ-mutant uveal melanoma cell lines with a Gαq/11 inhibitor, YM-254890, and conducted a kinase signaling proteomic screen using multiplexed-kinase inhibitors followed by mass spectrometry. We observed downregulated expression and/or activity of 22 kinases. A custom siRNA screen targeting these kinases demonstrated that knockdown of microtubule affinity regulating kinase 3 (MARK3) and serine/threonine kinase 10 (STK10) significantly reduced uveal melanoma cell growth and decreased expression of cell cycle proteins. Additionally, knockdown of MARK3 but not STK10 decreased ERK1/2 phosphorylation. Analysis of RNA-sequencing and proteomic data showed that Gαq signaling regulates STK10 expression and MARK3 activity. Our findings suggest an involvement of STK10 and MARK3 in the Gαq/11 oncogenic pathway and prompt further investigation into the specific roles and targeting potential of these kinases in uveal melanoma.

This open-labeled and comparative study aimed to test the efficacy and safety of a fermented rice extract-based substance containing yeast-fermented powder having aldehyde dehydrogenase (KisLip®, Pico Entech, Republic of Korea) in healthy male individuals. Healthy male subjects (n = 20) consumed 90 g of alcohol at their first visit. At the second visit, participants consumed 90 g of alcohol or alcohol with a low dose of KISLip® (2000 mg, KL-L) and then 90 g of alcohol or alcohol with a high dose of KISLip® (3000 mg, KL-H) at the third visit. The efficacy of KISLip® depends on the mutational status of important genes related to alcohol metabolism, including alcohol dehydrogenase (ADH1B), cytochrome P4502E1 (CYP2E1 (5B) and CYP2E1 (6)), and aldehyde dehydrogenase (ALDH2). KISLip® significantly reduced the highest level (Cmax) of alcohol and overall levels of acetaldehyde compared to the alcohol-only group in a dose-dependent manner. These significant effects of KISLip® on alcohol metabolism were observed independent of mutations in the four genes. In addition, hangover symptoms were significantly decreased in the KISLip® treated groups. During the study, the participants did not show any adverse events after KISLip® intake. This clinical study suggested that supplementation of KISLip® had beneficial effects on alcohol metabolism and might ameliorate the severity of hangovers without any adverse events. [ABSTRACT FROM AUTHOR]

Background and purpose: NCAPD2 is a protein-coding gene that plays a key role in cell proliferation, such as mitosis process. This study aimed to investigate the expression level and clinical value of NCAPD2 in glioma. Methods: The expression of NCAPD2 mRNA in various malignant tumor tissues was analyzed by GEPIA database. The expression level of NCAPD2 mRNA in glioma was analyzed by Ualcan, UCSC Xena and TIMER2.0 database. Immunohistochemical S-P method was used to detect and analyze the expression of NCAPD2 protein in normal brain tissue and glioma tissue, as well as the relationship between the overexpression of NCAPD2 protein and the clinicopathological features of glioma. The expression level of NCAPD2 in glioma cells was verified by Western blot. LinkedOmics database was used to explore genes co-expressed with NCAPD2. Metascape database and GoPlot database were used for enrichment analysis. Results: Database analysis showed that the expression of NCAPD2 was higher in various malignant tissues than in normal tissues (P ＜ 0.05), and the expression level of NCAPD2 was higher in isocitrate dehydrogenase 1 (IDH1) mutant-type glioma than in IDH1 wild-type glioma (P ＜ 0.05). Immunohistochemical results showed that NCAPD2 protein was mainly expressed in the nucleus and cytoplasm, and the positive expression rate and strong positive expression rate of NCAPD2 protein in glioma tissues were significantly higher than those in normal brain tissues (P ＜ 0.01). However, NCAPD2 protein was positively correlated with clinical classification of glioma patients (P ＜ 0.05). Western blot assay showed that the expression of NCAPD2 protein in glioma cells was significantly higher than in glial cells (P ＜ 0.05). GO enrichment analysis showed that NCAPD2 gene was mainly related to cell cycle progression (P ＜ 0.01). Conclusion: The overexpression of NCAPD2 in glioma is closely related to clinical grade and cell cycle progression, which indicates that NCAPD2 plays an important role in the occurrence and development of glioma.

Uveal melanoma (UM) is the deadliest form of eye cancer in adults. Inactivating mutations and/or loss of expression of the gene encoding BRCA1-associated protein 1 (BAP1) in UM tumors are associated with an increased risk of metastasis. To investigate the mechanisms underlying this risk, we explored the functional consequences of BAP1 deficiency. UM cell lines expressing mutant BAP1 grew more slowly than those expressing wild-type BAP1 in culture and in vivo. The ability of BAP1 reconstitution to restore cell proliferation in BAP1-deficient cells required its deubiquitylase activity. Proteomic analysis showed that BAP1-deficient cells had decreased phosphorylation of ribosomal S6 and its upstream regulator, p70S6K1, compared with both wild-type and BAP1 reconstituted cells. In turn, expression of p70S6K1 increased S6 phosphorylation and proliferation of BAP1-deficient UM cells. Consistent with these findings, BAP1 mutant primary UM tumors expressed lower amounts of p70S6K1 target genes, and S6 phosphorylation was decreased in BAP1 mutant patient-derived xenografts (PDXs), which grew more slowly than wild-type PDXs in the liver (the main metastatic site of UM) in mice. BAP1-deficient UM cells were also more resistant to amino acid starvation, which was associated with diminished phosphorylation of S6. These studies demonstrate that BAP1 deficiency slows the proliferation of UM cells through regulation of S6 phosphorylation. These characteristics may be associated with metastasis by ensuring survival during amino acid starvation. Editor's summary: Deficiency in the protein BAP1 is associated with increased metastatic progression of uveal melanoma (UM), an intraocular tumor. Chua et al. explored this connection in UM cell lines and patient samples. Loss of BAP1 or its deubiquitylase function decreased proliferation and signaling mediated by the protein S6 in UM cells and tumors. This loss of S6 signaling correlated with increased cell survival during amino acid starvation. Because nutrient deficiency is a type of stress common to the metastatic process, these findings suggest that BAP1 deficiency may support the survival of UM cells during metastasis. —Leslie K. Ferrarelli [ABSTRACT FROM AUTHOR]