Low-dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the DNA methylation-mediated MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway in acute myeloid leukemia
Abstract Background Ferroptosis is a new form of nonapoptotic and iron-dependent type of cell death. Glutathione peroxidase-4 (GPX4) plays an essential role in anti-ferroptosis by reducing lipid peroxidation. Although acute myeloid leukemia (AML) cells, especially relapsed and refractory (R/R)-AML,...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-02-01
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| Series: | Experimental Hematology & Oncology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40164-024-00489-4 |
| _version_ | 1797275556520132608 |
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| author | Shuya Feng Yigang Yuan Zihan Lin Min Li Daijiao Ye Liuzhi Shi Danyang Li Min Zhao Chen Meng Xiaofei He Shanshan Wu Fang Xiong Siyu Ye Junjun Yang Haifeng Zhuang Lili Hong Shenmeng Gao |
| author_facet | Shuya Feng Yigang Yuan Zihan Lin Min Li Daijiao Ye Liuzhi Shi Danyang Li Min Zhao Chen Meng Xiaofei He Shanshan Wu Fang Xiong Siyu Ye Junjun Yang Haifeng Zhuang Lili Hong Shenmeng Gao |
| author_sort | Shuya Feng |
| collection | DOAJ |
| description | Abstract Background Ferroptosis is a new form of nonapoptotic and iron-dependent type of cell death. Glutathione peroxidase-4 (GPX4) plays an essential role in anti-ferroptosis by reducing lipid peroxidation. Although acute myeloid leukemia (AML) cells, especially relapsed and refractory (R/R)-AML, present high GPX4 levels and enzyme activities, pharmacological inhibition of GPX4 alone has limited application in AML. Thus, whether inhibition of GPX4 combined with other therapeutic reagents has effective application in AML is largely unknown. Methods Lipid reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH) assays were used to assess ferroptosis in AML cells treated with the hypomethylating agent (HMA) decitabine (DAC), ferroptosis-inducer (FIN) RAS-selective lethal 3 (RSL3), or their combination. Combination index (CI) analysis was used to assess the synergistic activity of DAC + RSL3 against AML cells. Finally, we evaluated the synergistic activity of DAC + RSL3 in murine AML and a human R/R-AML-xenografted NSG model in vivo. Results We first assessed GPX4 expression and found that GPX4 levels were higher in AML cells, especially those with MLL rearrangements, than in NCs. Knockdown of GPX4 by shRNA and indirect inhibition of GPX4 enzyme activity by RSL3 robustly induced ferroptosis in AML cells. To reduce the dose of RSL3 and avoid side effects, low doses of DAC (0.5 µM) and RSL3 (0.05 µM) synergistically facilitate ferroptosis by inhibiting the AMP-activated protein kinase (AMPK)-SLC7A11-GPX4 axis. Knockdown of AMPK by shRNA enhanced ferroptosis, and overexpression of SLC7A11 and GPX4 rescued DAC + RSL3-induced anti-leukemogenesis. Mechanistically, DAC increased the expression of MAGEA6 by reducing MAGEA6 promoter hypermethylation. Overexpression of MAGEA6 induced the degradation of AMPK, suggesting that DAC inhibits the AMPK-SLC7A11-GPX4 axis by increasing MAGEA6 expression. In addition, DAC + RSL3 synergistically reduced leukemic burden and extended overall survival compared with either DAC or RSL3 treatment in the MLL-AF9-transformed murine model. Finally, DAC + RSL3 synergistically reduced viability in untreated and R/R-AML cells and extended overall survival in two R/R-AML-xenografted NSG mouse models. Conclusions Our study first identify vulnerability to ferroptosis by regulating MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway. Combined treatment with HMAs and FINs provides a potential therapeutic choice for AML patients, especially for R/R-AML. |
| first_indexed | 2024-03-07T15:16:10Z |
| format | Article |
| id | doaj.art-84b5eef75bfa4328af9bf045a1b26690 |
| institution | Directory Open Access Journal |
| issn | 2162-3619 |
| language | English |
| last_indexed | 2024-03-07T15:16:10Z |
| publishDate | 2024-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Experimental Hematology & Oncology |
| spelling | doaj.art-84b5eef75bfa4328af9bf045a1b266902024-03-05T17:54:44ZengBMCExperimental Hematology & Oncology2162-36192024-02-0113112010.1186/s40164-024-00489-4Low-dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the DNA methylation-mediated MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway in acute myeloid leukemiaShuya Feng0Yigang Yuan1Zihan Lin2Min Li3Daijiao Ye4Liuzhi Shi5Danyang Li6Min Zhao7Chen Meng8Xiaofei He9Shanshan Wu10Fang Xiong11Siyu Ye12Junjun Yang13Haifeng Zhuang14Lili Hong15Shenmeng Gao16Medical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityDepartment of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityThe Children’s Hospital of Zhejiang University School of MedicineSchool of Marine Sciences, Ningbo UniversityDepartment of Laboratory Medicine, The Second Affiliated Hospital, Yuying Children’s Hospital of Wenzhou Medical UniversityDepartment of Clinical Hematology and Transfusion, The First Affiliated Hospital of Zhejiang Chinese Medical UniversityDepartment of Clinical Hematology and Transfusion, The First Affiliated Hospital of Zhejiang Chinese Medical UniversityMedical Research Center, The First Affiliated Hospital of Wenzhou Medical UniversityAbstract Background Ferroptosis is a new form of nonapoptotic and iron-dependent type of cell death. Glutathione peroxidase-4 (GPX4) plays an essential role in anti-ferroptosis by reducing lipid peroxidation. Although acute myeloid leukemia (AML) cells, especially relapsed and refractory (R/R)-AML, present high GPX4 levels and enzyme activities, pharmacological inhibition of GPX4 alone has limited application in AML. Thus, whether inhibition of GPX4 combined with other therapeutic reagents has effective application in AML is largely unknown. Methods Lipid reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH) assays were used to assess ferroptosis in AML cells treated with the hypomethylating agent (HMA) decitabine (DAC), ferroptosis-inducer (FIN) RAS-selective lethal 3 (RSL3), or their combination. Combination index (CI) analysis was used to assess the synergistic activity of DAC + RSL3 against AML cells. Finally, we evaluated the synergistic activity of DAC + RSL3 in murine AML and a human R/R-AML-xenografted NSG model in vivo. Results We first assessed GPX4 expression and found that GPX4 levels were higher in AML cells, especially those with MLL rearrangements, than in NCs. Knockdown of GPX4 by shRNA and indirect inhibition of GPX4 enzyme activity by RSL3 robustly induced ferroptosis in AML cells. To reduce the dose of RSL3 and avoid side effects, low doses of DAC (0.5 µM) and RSL3 (0.05 µM) synergistically facilitate ferroptosis by inhibiting the AMP-activated protein kinase (AMPK)-SLC7A11-GPX4 axis. Knockdown of AMPK by shRNA enhanced ferroptosis, and overexpression of SLC7A11 and GPX4 rescued DAC + RSL3-induced anti-leukemogenesis. Mechanistically, DAC increased the expression of MAGEA6 by reducing MAGEA6 promoter hypermethylation. Overexpression of MAGEA6 induced the degradation of AMPK, suggesting that DAC inhibits the AMPK-SLC7A11-GPX4 axis by increasing MAGEA6 expression. In addition, DAC + RSL3 synergistically reduced leukemic burden and extended overall survival compared with either DAC or RSL3 treatment in the MLL-AF9-transformed murine model. Finally, DAC + RSL3 synergistically reduced viability in untreated and R/R-AML cells and extended overall survival in two R/R-AML-xenografted NSG mouse models. Conclusions Our study first identify vulnerability to ferroptosis by regulating MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway. Combined treatment with HMAs and FINs provides a potential therapeutic choice for AML patients, especially for R/R-AML.https://doi.org/10.1186/s40164-024-00489-4FerroptosisGlutathione peroxidase-4AMPKAcute myeloid leukemiaHypomethylating agent |
| spellingShingle | Shuya Feng Yigang Yuan Zihan Lin Min Li Daijiao Ye Liuzhi Shi Danyang Li Min Zhao Chen Meng Xiaofei He Shanshan Wu Fang Xiong Siyu Ye Junjun Yang Haifeng Zhuang Lili Hong Shenmeng Gao Low-dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the DNA methylation-mediated MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway in acute myeloid leukemia Experimental Hematology & Oncology Ferroptosis Glutathione peroxidase-4 AMPK Acute myeloid leukemia Hypomethylating agent |
| title | Low-dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the DNA methylation-mediated MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway in acute myeloid leukemia |
| title_full | Low-dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the DNA methylation-mediated MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway in acute myeloid leukemia |
| title_fullStr | Low-dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the DNA methylation-mediated MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway in acute myeloid leukemia |
| title_full_unstemmed | Low-dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the DNA methylation-mediated MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway in acute myeloid leukemia |
| title_short | Low-dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the DNA methylation-mediated MAGEA6-AMPK-SLC7A11-GPX4 signaling pathway in acute myeloid leukemia |
| title_sort | low dose hypomethylating agents cooperate with ferroptosis inducers to enhance ferroptosis by regulating the dna methylation mediated magea6 ampk slc7a11 gpx4 signaling pathway in acute myeloid leukemia |
| topic | Ferroptosis Glutathione peroxidase-4 AMPK Acute myeloid leukemia Hypomethylating agent |
| url | https://doi.org/10.1186/s40164-024-00489-4 |
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