Marietta Zille1, Saravanan S Karuppagounder1, Yingxin Chen1, Peter J Gough1, John Bertin1, Joshua Finger1, Teresa A Milner1, Elizabeth A Jonas1, Rajiv R Ratan2. 1. From the Burke Medical Research Institute, White Plains, New York (M.Z., S.S.K., Y.C., R.R.R.); Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, New York (M.Z., S.S.K., Y.C., T.A.M., R.R.R.); Host Defense Discovery Performance Unit, Infectious Diseases Therapy Area Unit (P.J.G.) and Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area (J.B., J.F.), GlaxoSmithKline, Collegeville, PA; Laboratory of Neuroendocrinology, The Rockefeller University, New York (T.A.M.); and Department of Internal Medicine, Section of Endocrinology, Yale University, New Haven, CT (E.A.J.). 2. From the Burke Medical Research Institute, White Plains, New York (M.Z., S.S.K., Y.C., R.R.R.); Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, New York (M.Z., S.S.K., Y.C., T.A.M., R.R.R.); Host Defense Discovery Performance Unit, Infectious Diseases Therapy Area Unit (P.J.G.) and Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area (J.B., J.F.), GlaxoSmithKline, Collegeville, PA; Laboratory of Neuroendocrinology, The Rockefeller University, New York (T.A.M.); and Department of Internal Medicine, Section of Endocrinology, Yale University, New Haven, CT (E.A.J.). rrr2001@med.cornell.edu.
Abstract
BACKGROUND AND PURPOSE: Intracerebral hemorrhage leads to disability or death with few established treatments. Adverse outcomes after intracerebral hemorrhage result from irreversible damage to neurons resulting from primary and secondary injury. Secondary injury has been attributed to hemoglobin and its oxidized product hemin from lysed red blood cells. The aim of this study was to identify the underlying cell death mechanisms attributable to secondary injury by hemoglobin and hemin to broaden treatment options. METHODS: We investigated cell death mechanisms in cultured neurons exposed to hemoglobin or hemin. Chemical inhibitors implicated in all known cell death pathways were used. Identified cell death mechanisms were confirmed using molecular markers and electron microscopy. RESULTS: Chemical inhibitors of ferroptosis and necroptosis protected against hemoglobin- and hemin-induced toxicity. By contrast, inhibitors of caspase-dependent apoptosis, protein or mRNA synthesis, autophagy, mitophagy, or parthanatos had no effect. Accordingly, molecular markers of ferroptosis and necroptosis were increased after intracerebral hemorrhage in vitro and in vivo. Electron microscopy showed that hemin induced a necrotic phenotype. Necroptosis and ferroptosis inhibitors each abrogated death by >80% and had similar therapeutic windows in vitro. CONCLUSIONS: Experimental intracerebral hemorrhage shares features of ferroptotic and necroptotic cell death, but not caspase-dependent apoptosis or autophagy. We propose that ferroptosis or necroptotic signaling induced by lysed blood is sufficient to reach a threshold of death that leads to neuronal necrosis and that inhibition of either of these pathways can bring cells below that threshold to survival.
BACKGROUND AND PURPOSE:Intracerebral hemorrhage leads to disability or death with few established treatments. Adverse outcomes after intracerebral hemorrhage result from irreversible damage to neurons resulting from primary and secondary injury. Secondary injury has been attributed to hemoglobin and its oxidized product hemin from lysed red blood cells. The aim of this study was to identify the underlying cell death mechanisms attributable to secondary injury by hemoglobin and hemin to broaden treatment options. METHODS: We investigated cell death mechanisms in cultured neurons exposed to hemoglobin or hemin. Chemical inhibitors implicated in all known cell death pathways were used. Identified cell death mechanisms were confirmed using molecular markers and electron microscopy. RESULTS: Chemical inhibitors of ferroptosis and necroptosis protected against hemoglobin- and hemin-induced toxicity. By contrast, inhibitors of caspase-dependent apoptosis, protein or mRNA synthesis, autophagy, mitophagy, or parthanatos had no effect. Accordingly, molecular markers of ferroptosis and necroptosis were increased after intracerebral hemorrhage in vitro and in vivo. Electron microscopy showed that hemin induced a necrotic phenotype. Necroptosis and ferroptosis inhibitors each abrogated death by >80% and had similar therapeutic windows in vitro. CONCLUSIONS: Experimental intracerebral hemorrhage shares features of ferroptotic and necroptotic cell death, but not caspase-dependent apoptosis or autophagy. We propose that ferroptosis or necroptotic signaling induced by lysed blood is sufficient to reach a threshold of death that leads to neuronal necrosis and that inhibition of either of these pathways can bring cells below that threshold to survival.
Authors: Scott J Dixon; Kathryn M Lemberg; Michael R Lamprecht; Rachid Skouta; Eleina M Zaitsev; Caroline E Gleason; Darpan N Patel; Andras J Bauer; Alexandra M Cantley; Wan Seok Yang; Barclay Morrison; Brent R Stockwell Journal: Cell Date: 2012-05-25 Impact factor: 41.582
Authors: Charlotte Jj van Asch; Merel Ja Luitse; Gabriël Je Rinkel; Ingeborg van der Tweel; Ale Algra; Catharina Jm Klijn Journal: Lancet Neurol Date: 2010-01-05 Impact factor: 44.182
Authors: Nicholas Yagoda; Moritz von Rechenberg; Elma Zaganjor; Andras J Bauer; Wan Seok Yang; Daniel J Fridman; Adam J Wolpaw; Inese Smukste; John M Peltier; J Jay Boniface; Richard Smith; Stephen L Lessnick; Sudhir Sahasrabudhe; Brent R Stockwell Journal: Nature Date: 2007-06-14 Impact factor: 49.962
Authors: Scott B Berger; Viera Kasparcova; Sandy Hoffman; Barb Swift; Lauren Dare; Michelle Schaeffer; Carol Capriotti; Michael Cook; Joshua Finger; Angela Hughes-Earle; Philip A Harris; William J Kaiser; Edward S Mocarski; John Bertin; Peter J Gough Journal: J Immunol Date: 2014-05-12 Impact factor: 5.422
Authors: Saravanan S Karuppagounder; Ishraq Alim; Soah J Khim; Megan W Bourassa; Sama F Sleiman; Roseleen John; Cyrille C Thinnes; Tzu-Lan Yeh; Marina Demetriades; Sandra Neitemeier; Dana Cruz; Irina Gazaryan; David W Killilea; Lewis Morgenstern; Guohua Xi; Richard F Keep; Timothy Schallert; Ryan V Tappero; Jian Zhong; Sunghee Cho; Frederick R Maxfield; Theodore R Holman; Carsten Culmsee; Guo-Hua Fong; Yijing Su; Guo-li Ming; Hongjun Song; John W Cave; Christopher J Schofield; Frederick Colbourne; Giovanni Coppola; Rajiv R Ratan Journal: Sci Transl Med Date: 2016-03-02 Impact factor: 17.956
Authors: Sarah B Berman; Ying-bei Chen; Bing Qi; J Michael McCaffery; Edmund B Rucker; Sandra Goebbels; Klaus-Armin Nave; Beth A Arnold; Elizabeth A Jonas; Fernando J Pineda; J Marie Hardwick Journal: J Cell Biol Date: 2009-03-02 Impact factor: 10.539
Authors: Che-Lin Hu; Mara Nydes; Kara L Shanley; Itzy E Morales Pantoja; Tamara A Howard; Oscar A Bizzozero Journal: J Neurochem Date: 2018-12-03 Impact factor: 5.372