TY - JOUR
T1 - Hemin-Induced Death Models Hemorrhagic Stroke and Is a Variant of Classical Neuronal Ferroptosis
AU - Zille, Marietta
AU - Oses-Prieto, Juan A.
AU - Savage, Sara R.
AU - Karuppagounder, Saravanan S.
AU - Chen, Yingxin
AU - Kumar, Amit
AU - Morris, John H.
AU - Scheidt, Karl A.
AU - Burlingame, Alma L.
AU - Ratan, Rajiv R.
N1 - Funding Information:
This work was supported by the German Research Foundation Grant DFG Zi 1613/1-1 (to M.Z.), funding from the Sheldon G and Dr. Miriam Adelson Medical Research Foundation and the Sperling Center for Hemorrhagic Stroke Recovery, and the National Institutes of Health Grant P01 NIA AG014930, project 1 (to R.R.R.). The authors declare no competing financial interests. Correspondence should be addressed to Marietta Zille at marietta.zille@univie.ac.at or Rajiv R. Ratan at rrr2001@med.cornell.edu. https://doi.org/10.1523/JNEUROSCI.0923-20.2021 Copyright © 2022 the authors
Publisher Copyright:
Copyright © 2022 the authors
PY - 2022/3/9
Y1 - 2022/3/9
N2 - Ferroptosis is a caspase-independent, iron-dependent form of regulated necrosis extant in traumatic brain injury, Huntington disease, and hemorrhagic stroke. It can be activated by cystine deprivation leading to glutathione depletion, the insufficiency of the antioxidant glutathione peroxidase-4, and the hemolysis products hemoglobin and hemin. A cardinal feature of ferroptosis is extracellular signal-regulated kinase (ERK)1/2 activation culminating in its translocation to the nucleus. We have previously confirmed that the mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor U0126 inhibits persistent ERK1/2 phosphorylation and ferroptosis. Here, we show that hemin exposure, a model of secondary injury in brain hemorrhage and ferroptosis, activated ERK1/2 in mouse neurons. Accordingly, MEK inhibitor U0126 protected against hemin-induced ferroptosis. Unexpectedly, U0126 prevented hemin-induced ferroptosis independent of its ability to inhibit ERK1/2 signaling. In contrast to classical ferroptosis in neurons or cancer cells, chemically diverse inhibitors of MEK did not block hemin-induced ferroptosis, nor did the forced expression of the ERK-selective MAP kinase phosphatase (MKP)3. We conclude that hemin or hemoglobin-induced ferroptosis, unlike glutathione depletion, is ERK1/2-independent. Together with recent studies, our findings suggest the existence of a novel subtype of neuronal ferroptosis relevant to bleeding in the brain that is 5-lipoxygenase-dependent, ERK-independent, and transcription-independent. Remarkably, our unbiased phosphoproteome analysis revealed dramatic differences in phosphorylation induced by two ferroptosis subtypes. As U0126 also reduced cell death and improved functional recovery after hemorrhagic stroke in male mice, our analysis also provides a template on which to build a search for U0126’s effects in a variant of neuronal ferroptosis.
AB - Ferroptosis is a caspase-independent, iron-dependent form of regulated necrosis extant in traumatic brain injury, Huntington disease, and hemorrhagic stroke. It can be activated by cystine deprivation leading to glutathione depletion, the insufficiency of the antioxidant glutathione peroxidase-4, and the hemolysis products hemoglobin and hemin. A cardinal feature of ferroptosis is extracellular signal-regulated kinase (ERK)1/2 activation culminating in its translocation to the nucleus. We have previously confirmed that the mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor U0126 inhibits persistent ERK1/2 phosphorylation and ferroptosis. Here, we show that hemin exposure, a model of secondary injury in brain hemorrhage and ferroptosis, activated ERK1/2 in mouse neurons. Accordingly, MEK inhibitor U0126 protected against hemin-induced ferroptosis. Unexpectedly, U0126 prevented hemin-induced ferroptosis independent of its ability to inhibit ERK1/2 signaling. In contrast to classical ferroptosis in neurons or cancer cells, chemically diverse inhibitors of MEK did not block hemin-induced ferroptosis, nor did the forced expression of the ERK-selective MAP kinase phosphatase (MKP)3. We conclude that hemin or hemoglobin-induced ferroptosis, unlike glutathione depletion, is ERK1/2-independent. Together with recent studies, our findings suggest the existence of a novel subtype of neuronal ferroptosis relevant to bleeding in the brain that is 5-lipoxygenase-dependent, ERK-independent, and transcription-independent. Remarkably, our unbiased phosphoproteome analysis revealed dramatic differences in phosphorylation induced by two ferroptosis subtypes. As U0126 also reduced cell death and improved functional recovery after hemorrhagic stroke in male mice, our analysis also provides a template on which to build a search for U0126’s effects in a variant of neuronal ferroptosis.
KW - MAP signaling
KW - brain hemorrhage
KW - cell death
KW - ferroptosis
KW - phosphoproteomics
KW - stroke
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U2 - 10.1523/JNEUROSCI.0923-20.2021
DO - 10.1523/JNEUROSCI.0923-20.2021
M3 - Article
C2 - 34987108
AN - SCOPUS:85126152628
SN - 0270-6474
VL - 42
SP - 2065
EP - 2079
JO - Journal of Neuroscience
JF - Journal of Neuroscience
IS - 10
ER -