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bioRxiv · 10.1101/2024.02.22.581432

NINJ1 regulates ferroptosis via xCT antiporter interaction and CoA modulation

Chen, S.-Y.·Lin, C.-C.·Wu, J.·Chen, Y.·Wang, Y.-E.·Setayeshpour, Y.·Mestre, A.·Chi, J.-T. A.
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Abstract

Ninjurin-1 (NINJ1), initially identified as a stress-induced protein in neurons, recently emerged as a key mediator of plasma membrane rupture during apoptosis, necrosis, and pyroptosis. However, its involvement in ferroptosis remains unknown. Here, we demonstrate that NINJ1 also plays a crucial role in ferroptosis, but through a distinct mechanism. NINJ1 knockdown significantly protected cancer cells against ferroptosis induced by xCT inhibitors but no other classes of ferroptosis-inducing compounds (FINs). Glycine, known to inhibit canonical NINJ1-mediated membrane rupture in other cell deaths, had no impact on ferroptosis. A compound screen revealed that NINJ1-mediated ferroptosis protection can be abolished by pantothenate kinase inhibitor (PANKi), buthionine sulfoximine (BSO), and diethylmaleate (DEM). These results suggest that this ferroptosis protection is mediated via Coenzyme A (CoA) and glutathione (GSH), both of which were found to be elevated upon NINJ1 knockdown. Furthermore, we discovered that NINJ1 interacts with the xCT antiporter, which is responsible for cystine uptake for the biosynthesis of CoA and GSH. The removal of NINJ1 increased xCT levels and stability, enhanced cystine uptake, and contributed to elevated CoA and GSH levels, collectively contributing to ferroptosis protection. These findings reveal that NINJ1 regulates ferroptosis via a non-canonical mechanism, distinct from other regulated cell deaths. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/581432v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@9fa073org.highwire.dtl.DTLVardef@1df0137org.highwire.dtl.DTLVardef@1c8e8cborg.highwire.dtl.DTLVardef@12b76b3_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Chen, S.-Y., Lin, C.-C., Wu, J., Chen, Y., Wang, Y.-E., Setayeshpour, Y., Mestre, A., Chi, J.-T. A.. 2024-02-27. NINJ1 regulates ferroptosis via xCT antiporter interaction and CoA modulation. https://doi.org/10.1101/2024.02.22.581432

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