Neuron-Specific Tumor Necrosis Factor Receptor-Associated Factor 3 Is a Central Regulator of Neuronal Death in Acute Ischemic Stroke.
Neuronal death after ischemic stroke involves multiple pathophysiological events, as well as a complex molecular mechanism. Inhibiting a single therapeutic target that is involved in several ischemic signaling cascades may be a promising strategy for stroke management. Here, we report the versatile biological roles of tumor necrosis factor receptor-associated factor 3 (TRAF3) in ischemic stroke. Using several genetically manipulated mouse strains, we also demonstrated that TRAF3 inhibition can be neuroprotective. TRAF3 expression, which is robustly induced in response to ischemia/reperfusion (I/R) injury, was detected in neurons. Overexpression of TRAF3 in neurons led to aggravated neuronal loss and enlarged infarcts; these effects were reversed in TRAF3-knockout mice. Neuronal TRAF3 also contributed to c-Jun kinase-, nuclear factor κB- and Rac-1-induced neuronal death, inflammation, and oxidative stress. Mechanistically, we showed that TRAF3 interacts with transforming growth factor-β-activated kinase 1 (TAK1) and potentiates phosphorylation and activation of TAK1. Phosphorylated TAK1 sequentially initiated activation of nuclear factor κB, Rac-1/NADPH oxidase, and c-Jun kinase/c-Jun signaling cascades. Using a combination of adenoviruses encoding dominant-negative TAK1 and the TAK1 inhibitor 5Z-7-oxozeaenol, we demonstrated that the TRAF3-mediated activation of ischemic cascades was TAK1-dependent. More importantly, the adverse phenotypes observed in TRAF3-overexpressing mice were completely reversed when the TRAF3-TAK1 interaction was prevented. Therefore, we have shown that TRAF3 is a central regulator of ischemic pathways, including nuclear factor κB, Rac-1, and c-Jun kinase signaling, via its interaction with and activation of TAK1. Furthermore, certain components of the TRAF3-TAK1 signaling pathway are potentially promising therapeutic targets in ischemic stroke.
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Jun
J
Gong
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Zuo-Zhi
ZZ
Li
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Sen
S
Guo
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Xiao-Jing
XJ
Zhang
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Peng
P
Zhang
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Guang-Nian
GN
Zhao
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Lu
L
Gao
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Yan
Y
Zhang
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Ankang
A
Zheng
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Xiao-Fei
XF
Zhang
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.).
Mei
M
Xiang
From the Department of Cardiology, Renmin Hospital (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), Animal Experiment Center/Animal Biosafety Level-III Laboratory (J.G., S.G., X.-J.Z., P.Z., G.-N.Z., Y.Z., A.Z., M.X., H.L.), and College of Life Sciences (X.-F.Z.), Wuhan University, Wuhan, China; Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China (Z.-Z.L.); National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-Z.L.); and Department of Cardiology, Institute of Cardiovascular Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (L.G.). lihl@whu.edu.cn.
Hongliang
H
Li
D000818:Animals; D002545:Brain Ischemia / Q000378:metabolism* / Q000473:pathology; D016923:Cell Death / Q000502:physiology*; D020930:MAP Kinase Kinase Kinases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D048055:Mitogen-Activated Protein Kinase 8 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D015398:Signal Transduction / Q000502:physiology; D020521:Stroke / Q000378:metabolism* / Q000473:pathology; D048008:TNF Receptor-Associated Factor 3 / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation; D000099151:MAP Kinase Kinase Kinase 7
D020930:MAP Kinase Kinase Kinases; D048055:Mitogen-Activated Protein Kinase 8; D016328:NF-kappa B; D048008:TNF Receptor-Associated Factor 3; D000099151:MAP Kinase Kinase Kinase 7
TAK1 MAPKKK; TNF receptor-associated factor 3; inflammation; oxidative stress; stroke