Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis
Small interfering RNAs (siRNAs) are widely used in biomedical research and in clinical trials. Here, we demonstrate that siRNA treatment is commonly associated with significant sensitization to ferroptosis, independently of the target protein knockdown. Genetically targeting mitochondrial antiviral-...
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sg-ntu-dr.10356-1783902024-06-23T15:38:37Z Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis von Mässenhausen, Anne Schlecht, Marlena Nastassja Beer, Kristina Maremonti, Francesca Tonnus, Wulf Belavgeni, Alexia Gavali, Shubhangi Flade, Karolin Riley, Joel S. Zamora Gonzalez, Nadia Brucker, Anne Becker, Jorunn Naila Tmava, Mirela Meyer, Claudia Peitzsch, Mirko Hugo, Christian Gembardt, Florian Angeli, Jose Pedro Friedmann Bornstein, Stefan R. Tait, Stephen W. G. Linkermann, Andreas Lee Kong Chian School of Medicine (LKCMedicine) Medicine, Health and Life Sciences Signalling proteins Small interfering RNA Small interfering RNAs (siRNAs) are widely used in biomedical research and in clinical trials. Here, we demonstrate that siRNA treatment is commonly associated with significant sensitization to ferroptosis, independently of the target protein knockdown. Genetically targeting mitochondrial antiviral-signaling protein (MAVS) reversed the siRNA-mediated sensitizing effect, but no activation of canonical MAVS signaling, which involves phosphorylation of IkBα and interferon regulatory transcription factor 3 (IRF3), was observed. In contrast, MAVS mediated a noncanonical signal resulting in a prominent increase in mitochondrial ROS levels, and increase in the BACH1/pNRF2 transcription factor ratio and GPX4 up-regulation, which was associated with a 50% decrease in intracellular glutathione levels. We conclude that siRNAs commonly sensitize to ferroptosis and may severely compromise the conclusions drawn from silencing approaches in biomedical research. Finally, as ferroptosis contributes to a variety of pathophysiological processes, we cannot exclude side effects in human siRNA-based therapeutical concepts that should be clinically tested. Published version Work in the Linkermann Lab was funded by the German Research Foundation SFB-TRR205, SFB-TRR 127, and SPP3206, and a Heisenberg-Professorship to A.L., project number 324141047, the international research training group (IRTG) 2251, and the DFG grant 522190184. It was further supported by the BMBF (FERROPath consortium). SWGT is supported by a Cancer Research UK Programme Grant (DRCNPG-Jun22\100011). Furthermore, SWGT consults for Exo Therapeutics. 2024-06-18T01:54:26Z 2024-06-18T01:54:26Z 2024 Journal Article von Mässenhausen, A., Schlecht, M. N., Beer, K., Maremonti, F., Tonnus, W., Belavgeni, A., Gavali, S., Flade, K., Riley, J. S., Zamora Gonzalez, N., Brucker, A., Becker, J. N., Tmava, M., Meyer, C., Peitzsch, M., Hugo, C., Gembardt, F., Angeli, J. P. F., Bornstein, S. R., ...Linkermann, A. (2024). Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis. Science Advances, 10(11), eadk7329-. https://dx.doi.org/10.1126/sciadv.adk7329 2375-2548 https://hdl.handle.net/10356/178390 10.1126/sciadv.adk7329 38489367 2-s2.0-85188201656 11 10 eadk7329 en Science Advances © 2024 the Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a creative commons Attribution license 4.0 (CC BY). application/pdf |
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Medicine, Health and Life Sciences Signalling proteins Small interfering RNA |
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Medicine, Health and Life Sciences Signalling proteins Small interfering RNA von Mässenhausen, Anne Schlecht, Marlena Nastassja Beer, Kristina Maremonti, Francesca Tonnus, Wulf Belavgeni, Alexia Gavali, Shubhangi Flade, Karolin Riley, Joel S. Zamora Gonzalez, Nadia Brucker, Anne Becker, Jorunn Naila Tmava, Mirela Meyer, Claudia Peitzsch, Mirko Hugo, Christian Gembardt, Florian Angeli, Jose Pedro Friedmann Bornstein, Stefan R. Tait, Stephen W. G. Linkermann, Andreas Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis |
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Small interfering RNAs (siRNAs) are widely used in biomedical research and in clinical trials. Here, we demonstrate that siRNA treatment is commonly associated with significant sensitization to ferroptosis, independently of the target protein knockdown. Genetically targeting mitochondrial antiviral-signaling protein (MAVS) reversed the siRNA-mediated sensitizing effect, but no activation of canonical MAVS signaling, which involves phosphorylation of IkBα and interferon regulatory transcription factor 3 (IRF3), was observed. In contrast, MAVS mediated a noncanonical signal resulting in a prominent increase in mitochondrial ROS levels, and increase in the BACH1/pNRF2 transcription factor ratio and GPX4 up-regulation, which was associated with a 50% decrease in intracellular glutathione levels. We conclude that siRNAs commonly sensitize to ferroptosis and may severely compromise the conclusions drawn from silencing approaches in biomedical research. Finally, as ferroptosis contributes to a variety of pathophysiological processes, we cannot exclude side effects in human siRNA-based therapeutical concepts that should be clinically tested. |
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Lee Kong Chian School of Medicine (LKCMedicine) |
author_facet |
Lee Kong Chian School of Medicine (LKCMedicine) von Mässenhausen, Anne Schlecht, Marlena Nastassja Beer, Kristina Maremonti, Francesca Tonnus, Wulf Belavgeni, Alexia Gavali, Shubhangi Flade, Karolin Riley, Joel S. Zamora Gonzalez, Nadia Brucker, Anne Becker, Jorunn Naila Tmava, Mirela Meyer, Claudia Peitzsch, Mirko Hugo, Christian Gembardt, Florian Angeli, Jose Pedro Friedmann Bornstein, Stefan R. Tait, Stephen W. G. Linkermann, Andreas |
format |
Article |
author |
von Mässenhausen, Anne Schlecht, Marlena Nastassja Beer, Kristina Maremonti, Francesca Tonnus, Wulf Belavgeni, Alexia Gavali, Shubhangi Flade, Karolin Riley, Joel S. Zamora Gonzalez, Nadia Brucker, Anne Becker, Jorunn Naila Tmava, Mirela Meyer, Claudia Peitzsch, Mirko Hugo, Christian Gembardt, Florian Angeli, Jose Pedro Friedmann Bornstein, Stefan R. Tait, Stephen W. G. Linkermann, Andreas |
author_sort |
von Mässenhausen, Anne |
title |
Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis |
title_short |
Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis |
title_full |
Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis |
title_fullStr |
Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis |
title_full_unstemmed |
Treatment with siRNAs is commonly associated with GPX4 up-regulation and target knockdown-independent sensitization to ferroptosis |
title_sort |
treatment with sirnas is commonly associated with gpx4 up-regulation and target knockdown-independent sensitization to ferroptosis |
publishDate |
2024 |
url |
https://hdl.handle.net/10356/178390 |
_version_ |
1814047380924792832 |