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Title:
CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma | Molecular Cancer
Description:
Background Hepatocellular carcinoma (HCC) is among the most common forms of cancer and is associated with poor patient outcomes. The emergence of therapeutic resistance has hampered the efficacy of targeted treatments employed to treat HCC patients to date. In this study, we conducted a series of CRISPR/Cas9 screens to identify genes associated with synthetic lethality capable of improving HCC patient clinical responses. Methods CRISPR-based loss-of-function genetic screens were used to target 18,053 protein-coding genes in HCC cells to identify chemotherapy-related synthetic lethal genes in these cells. Synergistic effects were analyzed through in vitro and in vivo analyses, while related mechanisms were explored through RNA-seq and metabolomics analyses. Potential inhibitors of identified genetic targets were selected through high-throughput virtual screening. Results The inhibition of phosphoseryl-tRNA kinase (PSTK) was found to increase HCC cell sensitivity to chemotherapeutic treatment. PSTK was associated with the suppression of chemotherapy-induced ferroptosis in HCC cells, and the depletion of PSTK resulted in the inactivation of glutathione peroxidative 4 (GPX4) and the disruption of glutathione (GSH) metabolism owing to the inhibition of selenocysteine and cysteine synthesis, thus enhancing the induction of ferroptosis upon targeted chemotherapeutic treatment. Punicalin, an agent used to treat hepatitis B virus (HBV), was identified as a possible PSTK inhibitor that exhibited synergistic efficacy when applied together with Sorafenib to treat HCC in vitro and in vivo. Conclusions These results highlight a key role for PSTK as a mediator of resistance to targeted therapeutic treatment in HCC cells that functions by suppressing ferroptotic induction. PSTK inhibitors may thus represent ideal candidates for overcoming drug resistance in HCC.
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Keywords {π}
pstk, hcc, cells, fig, cell, ferroptosis, gpx, article, treatment, pubmed, sorafenib, knockout, genes, levels, resistance, google, scholar, cas, cancer, expression, analyses, tumor, pstkko, samples, targeted, lines, results, gsh, significantly, punicalin, death, inhibitors, selenocysteine, therapy, erastin, central, hepatocellular, carcinoma, biosynthesis, groups, patients, screening, inhibition, induction, significant, abemaciclib, control, study, identified, treated,
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powerful gene-editing technology punicalin-induced pstk/gpx4 deficiency crispr/cas9 knockout screens dual-sgrna plasmid construct article download pdf genotype-specific cancer liabilities s-appt induces ferroptosis kaplan-meier curves demonstrating high-throughput functional genomics pstk-nc/pstk-ko cells punicalin-induced cell death high-throughput virtual screening resultant hep3b-cas9 cells immunohistochemistry auto-stainer approach specifically phosphorylates seryl-trna o-phosphoseryl-trna kinase pstk-regulated cell death structure-based virtual screen crispr/cas9 screens nature reviews genetics ferroptosis-induced tumor eradication sorafenib-induced cell death pstk/sod2 high/medium expression pan-cancer tcga analysis ifn-Ξ³-induced ferroptosis identified phosphoseryl-trna kinase xenograft-bearing mice treated targeted therapy-induced ferroptosis punicalin/geraniin/vehicle treated mice punicalin/geraniin/vehicle treatment groups protein-protein interaction networks paired t-test key ferroptosis-related genes approved multi-kinase inhibitor pstk-ko hcc cells pstk-sgrna1-sgrna2 construct pstk-specific sgrna sequences hep3b-pstk-ko cells pstk-ko hep3b cells genome-wide mapping hcc-specific therapeutic target treating pstk-ko cells full access crispr screening results generated pstk-ko cells recovering pstk/gpx4 expressions pstk slightly inhibited selenocysteine-dependent regulatory mechanism increasing abemaciclib/sorafenib concentrations control sgrna-transfected cells
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- CDK4/6 inhibition and sorafenib: a menage a deux in HCC therapy?
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headline:CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma
description:Hepatocellular carcinoma (HCC) is among the most common forms of cancer and is associated with poor patient outcomes. The emergence of therapeutic resistance has hampered the efficacy of targeted treatments employed to treat HCC patients to date. In this study, we conducted a series of CRISPR/Cas9 screens to identify genes associated with synthetic lethality capable of improving HCC patient clinical responses. CRISPR-based loss-of-function genetic screens were used to target 18,053 protein-coding genes in HCC cells to identify chemotherapy-related synthetic lethal genes in these cells. Synergistic effects were analyzed through in vitro and in vivo analyses, while related mechanisms were explored through RNA-seq and metabolomics analyses. Potential inhibitors of identified genetic targets were selected through high-throughput virtual screening. The inhibition of phosphoseryl-tRNA kinase (PSTK) was found to increase HCC cell sensitivity to chemotherapeutic treatment. PSTK was associated with the suppression of chemotherapy-induced ferroptosis in HCC cells, and the depletion of PSTK resulted in the inactivation of glutathione peroxidative 4 (GPX4) and the disruption of glutathione (GSH) metabolism owing to the inhibition of selenocysteine and cysteine synthesis, thus enhancing the induction of ferroptosis upon targeted chemotherapeutic treatment. Punicalin, an agent used to treat hepatitis B virus (HBV), was identified as a possible PSTK inhibitor that exhibited synergistic efficacy when applied together with Sorafenib to treat HCC in vitro and in vivo. These results highlight a key role for PSTK as a mediator of resistance to targeted therapeutic treatment in HCC cells that functions by suppressing ferroptotic induction. PSTK inhibitors may thus represent ideal candidates for overcoming drug resistance in HCC.
datePublished:2022-01-04T00:00:00Z
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Hepatocellular carcinoma
CRISPR library screening
PSTK
Ferroptosis
Cancer Research
Oncology
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headline:CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma
description:Hepatocellular carcinoma (HCC) is among the most common forms of cancer and is associated with poor patient outcomes. The emergence of therapeutic resistance has hampered the efficacy of targeted treatments employed to treat HCC patients to date. In this study, we conducted a series of CRISPR/Cas9 screens to identify genes associated with synthetic lethality capable of improving HCC patient clinical responses. CRISPR-based loss-of-function genetic screens were used to target 18,053 protein-coding genes in HCC cells to identify chemotherapy-related synthetic lethal genes in these cells. Synergistic effects were analyzed through in vitro and in vivo analyses, while related mechanisms were explored through RNA-seq and metabolomics analyses. Potential inhibitors of identified genetic targets were selected through high-throughput virtual screening. The inhibition of phosphoseryl-tRNA kinase (PSTK) was found to increase HCC cell sensitivity to chemotherapeutic treatment. PSTK was associated with the suppression of chemotherapy-induced ferroptosis in HCC cells, and the depletion of PSTK resulted in the inactivation of glutathione peroxidative 4 (GPX4) and the disruption of glutathione (GSH) metabolism owing to the inhibition of selenocysteine and cysteine synthesis, thus enhancing the induction of ferroptosis upon targeted chemotherapeutic treatment. Punicalin, an agent used to treat hepatitis B virus (HBV), was identified as a possible PSTK inhibitor that exhibited synergistic efficacy when applied together with Sorafenib to treat HCC in vitro and in vivo. These results highlight a key role for PSTK as a mediator of resistance to targeted therapeutic treatment in HCC cells that functions by suppressing ferroptotic induction. PSTK inhibitors may thus represent ideal candidates for overcoming drug resistance in HCC.
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Hepatocellular carcinoma
CRISPR library screening
PSTK
Ferroptosis
Cancer Research
Oncology
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