Phenytoin inhibits necroptosis

Anne von Mässenhausen (1) , Wulf Tonnus (1) , Nina Himmerkus , Simon Parmentier , Danish Saleh , Diego Rodriguez (2) , Jiraporn Ousingsawat , Rosalind L Ang , Joel M Weinberg , Ana B Sanz , Alberto Ortiz (3) , Adrian Zierleyn , Jan Ulrich Becker , Blandine Baratte (4, 5, 6, 7) , Nathalie Desban (6, 4) , Stéphane Bach (6, 4, 5, 7) , Ina Maria Schiessl , Shoko Nogusa , Siddharth Balachandran (8) , Hans Joachim Anders , Adrian T Ting , Markus Bleich , Alexei Degterev , Karl Kunzelmann , Stefan R Bornstein (9, 10, 11) , Douglas R Green , Christian Hugo , Andreas Linkermann (1)

1 Division of Nephrology, Medical Clinic, University Hospital Carl Gustav Carus, Technical University Dresden
2 UCM - Universidad Complutense de Madrid = Complutense University of Madrid [Madrid]
3 IIS‑Fundación Jiménez Diaz‑Autonoma University [Madrid, Spain]
4 LBI2M - Laboratoire de Biologie Intégrative des Modèles Marins
5 KISSf - Plate-forme de criblage d'inhibiteurs de protéines kinases=Kinase Inhibitor Specialized Screening facility
6 SBR - Station biologique de Roscoff = Roscoff Marine Station
7 FR2424 - Fédération de recherche de Roscoff
8 Fox Chase Cancer Center
9 TU Dresden - Technische Universität Dresden = Dresden University of Technology
10 King‘s College London
11 University hospital of Zurich [Zurich]

Receptor-interacting protein kinases 1 and 3 (RIPK1/3) have best been described for their role in mediating a regulated form of necrosis, referred to as necroptosis. During this process, RIPK3 phosphorylates mixed lineage kinase domainlike (MLKL) to cause plasma membrane rupture. RIPK3-deficient mice have recently been demonstrated to be protected in a series of disease models, but direct evidence for activation of necroptosis in vivo is still limited. Here, we sought to further examine the activation of necroptosis in kidney ischemia-reperfusion injury (IRI) and from TNFαinduced severe inflammatory response syndrome (SIRS), two models of RIPK3-dependent injury. In both models, MLKL-ko mice were significantly protected from injury to a degree that was slightly, but statistically significantly exceeding that of RIPK3-deficient mice. We also demonstrated, for the first time, accumulation of pMLKL in the necrotic tubules of human patients with acute kidney injury. However, our data also uncovered unexpected elevation of blood flow in MLKL-ko animals, which may be relevant to IRI and should be considered in the future. To further understand the mode of regulation of cell death by MLKL, we screened a panel of clinical plasma membrane channel blockers and we found phenytoin to inhibit necroptosis. However, we further found that phenytoin attenuated RIPK1 kinase activity in vitro, likely due to the hydantoin scaffold also present in necrostatin-1, and blocked upstream necrosome formation steps in the cells undergoing necroptosis. We further report that this clinically used anticonvulsant drug displayed protection from kidney IRI and TNFα-induces SIRS in vivo. Overall, our data reveal the relevance of RIPK3-pMLKL regulation for acute kidney injury and identifies an FDA-approved drug that may be useful for immediate clinical evaluation of inhibition of pro-death RIPK1/RIPK3 activities in human diseases.

Domaines: Sciences du Vivant [q-bio]; Médicaments; Pharmacologie

Anne von Mässenhausen, Wulf Tonnus, Nina Himmerkus, Simon Parmentier, Danish Saleh, et al.. Phenytoin inhibits necroptosis. Cell Death and Disease, 2018, 9 (3), pp.359. ⟨10.1038/s41419-018-0394-3⟩. ⟨hal-03871390⟩