DJ-1 suppresses ferroptosis through preserving the activity of S-adenosyl homocysteine hydrolase

Ferroptosis is a newly characterized form of regulated cell death mediated by iron-dependent accumulation of lipid reactive oxygen species and holds great potential for cancer therapy. However, the molecular mechanisms underlying ferroptosis remain largely elusive. In this study, we define an integrative role of DJ-1 in ferroptosis. Inhibition of DJ-1 potently enhances the sensitivity of tumor cells to ferroptosis inducers both in vitro and in vivo. Metabolic analysis and metabolite rescue assay reveal that DJ-1 depletion inhibits the transsulfuration pathway by disrupting the formation of the S-adenosyl homocysteine hydrolase tetramer and impairing its activity. Consequently, more ferroptosis is induced when homocysteine generation is decreased, which might be the only source of glutathione biosynthesis when cystine uptake is blocked. Thus, our findings show that DJ-1 determines the response of cancer cells to ferroptosis, and highlight a candidate therapeutic target to potentially improve the effect of ferroptosis-based antitumor therapy.


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Meidan Ying
Jan 30, 2020 1. Western Blot signal was recorded using a AI600 Imager (GE Healthcare Life Sciences). 2. Nano-LC-MS/MS data were acquired in Xcalibur 2.2 operation software (Thermo Fisher Scientific). 3. Flow cytometry data were collected using FACSuite flow cytometer (BD Biosciences). 4. EM Imaging data was collected using a transmission electron microscope software (Philips Electronic Instruments, Mahwah, USA). 5. Real-time PCR data was monitored using QuantStudio 6 Design and Analysis Software Version 2.3 (ThermoFisher). 6. Immunofluorescence Imaging data was collected using a Leica microscope software Leica Application Suite Version 4.3.0. 7. Fluorescent signal of SAHH activity assay was collected using Molecular Devices SpectraMax M5 plate reader software SoftMax Pro 6.3. 8. Metabolite analysis and quantitation were performed by the software Xcalibur 3.0.63 (Thermo Fisher Scientific). 9. The ELISA data analysis was collected using Molecular Devices SpectraMax M5 plate reader software SoftMax Pro 6.3.
1. The flow cytometry data were analysized using FlowJo Version 7.6 software. 2. All statistic analysis was conducted by Prism 5.0C (Graphpad Software) software (described in methods section) 1.The potential interaction proteins of DJ-1 based on BioID assay can be found in supplementary table (table S1) nature research | reporting summary

October 2018
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All studies must disclose on these points even when the disclosure is negative. Cell line source(s) 2. All the plasmids used in this manuscript including the ones in BioID assay will be available from the corresponding author upon reasonable request. 3. All the raw data of the WB was provided as supplementary dataset. 4. All original data for charts were provided as supplementary dataset.
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For the animal study, we don't show the group information in the cage labels and two experts were conducted. One is in charge of the administration and the other one is response for the data collection. The data collection person don't know the groups information until the end of the experiment. Thus, the investigators were blinded to group allocation during data collection and were not blinded to data analysis.