CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma
Abstract 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 ser...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-01-01
|
| Series: | Molecular Cancer |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12943-021-01466-9 |
| _version_ | 1798034391025319936 |
|---|---|
| author | Yiran Chen Li Li Jie Lan Yang Cui Xiaosong Rao Jing Zhao Tao Xing Gaoda Ju Guangtao Song Jizhong Lou Jun Liang |
| author_facet | Yiran Chen Li Li Jie Lan Yang Cui Xiaosong Rao Jing Zhao Tao Xing Gaoda Ju Guangtao Song Jizhong Lou Jun Liang |
| author_sort | Yiran Chen |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-04-11T20:42:45Z |
| format | Article |
| id | doaj.art-a8bfd4544ab640f68cf1709d5f21ff5a |
| institution | Directory Open Access Journal |
| issn | 1476-4598 |
| language | English |
| last_indexed | 2024-04-11T20:42:45Z |
| publishDate | 2022-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Cancer |
| spelling | doaj.art-a8bfd4544ab640f68cf1709d5f21ff5a2022-12-22T04:04:08ZengBMCMolecular Cancer1476-45982022-01-0121111710.1186/s12943-021-01466-9CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinomaYiran Chen0Li Li1Jie Lan2Yang Cui3Xiaosong Rao4Jing Zhao5Tao Xing6Gaoda Ju7Guangtao Song8Jizhong Lou9Jun Liang10Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Oncology, Peking University Cancer Hospital and InstituteDepartment of Oncology, Peking University International HospitalLaboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesLaboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesDepartment of Pathology, Boao Evergrande International HospitalDepartment of Medical Oncology, The Affiliated Hospital of Qingdao University, Qingdao UniversityKey Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Oncology, Peking University Cancer Hospital and InstituteKey Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Oncology, Peking University Cancer Hospital and InstituteLaboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesLaboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesKey Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Oncology, Peking University Cancer Hospital and InstituteAbstract 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.https://doi.org/10.1186/s12943-021-01466-9Hepatocellular carcinomaCRISPR library screeningPSTKFerroptosis |
| spellingShingle | Yiran Chen Li Li Jie Lan Yang Cui Xiaosong Rao Jing Zhao Tao Xing Gaoda Ju Guangtao Song Jizhong Lou Jun Liang CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma Molecular Cancer Hepatocellular carcinoma CRISPR library screening PSTK Ferroptosis |
| title | CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma |
| title_full | CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma |
| title_fullStr | CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma |
| title_full_unstemmed | CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma |
| title_short | CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma |
| title_sort | crispr screens uncover protective effect of pstk as a regulator of chemotherapy induced ferroptosis in hepatocellular carcinoma |
| topic | Hepatocellular carcinoma CRISPR library screening PSTK Ferroptosis |
| url | https://doi.org/10.1186/s12943-021-01466-9 |
| work_keys_str_mv | AT yiranchen crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT lili crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT jielan crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT yangcui crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT xiaosongrao crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT jingzhao crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT taoxing crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT gaodaju crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT guangtaosong crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT jizhonglou crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma AT junliang crisprscreensuncoverprotectiveeffectofpstkasaregulatorofchemotherapyinducedferroptosisinhepatocellularcarcinoma |