High-Content Genome-Wide RNAi Screen RevealsCCR3as a Key Mediator of Neuronal Cell Death.
Neuronal loss caused by ischemic injury, trauma, or disease can lead to devastating consequences for the individual. With the goal of limiting neuronal loss, a number of cell death pathways have been studied, but there may be additional contributors to neuronal death that are yet unknown. To identify previously unknown cell death mediators, we performed a high-content genome-wide screening of short, interfering RNA (siRNA) with an siRNA library in murine neural stem cells after exposure toN-methyl-N-nitroso-N'-nitroguanidine (MNNG), which leads to DNA damage and cell death. Eighty genes were identified as key mediators for cell death. Among them, 14 are known cell death mediators and 66 have not previously been linked to cell death pathways. Using an integrated approach with functional and bioinformatics analysis, we provide possible molecular networks, interconnected pathways, and/or protein complexes that may participate in cell death. Of the 66 genes, we selectedCCR3for further evaluation and found that CCR3 is a mediator of neuronal injury. CCR3 inhibition or deletion protects murine cortical cultures from oxygen-glucose deprivation-induced cell death, andCCR3deletion in mice provides protection from ischemia in vivo. Taken together, our findings suggest that CCR3 is a previously unknown mediator of cell death. Future identification of the neural cell death network in which CCR3 participates will enhance our understanding of the molecular mechanisms of neural cell death.
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Immunology, Neuroscience Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, State Key Laboratory of Medical Molecular Biology, Beijing, 100005, China.
Jianmin
0000-0003-4765-0812
J
Zhang
Department of Immunology, Neuroscience Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, State Key Laboratory of Medical Molecular Biology , Beijing, 100005, China.
Huaishan
H
Wang
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Omar
O
Sherbini
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Emily
E
Ling-Lin Pai
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Sung-Ung
0000-0002-8393-2588
SU
Kang
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Ji-Sun
JS
Kwon
Department of Immunology, Neuroscience Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, State Key Laboratory of Medical Molecular Biology , Beijing, 100005, China.
Jia
0000-0002-3992-4070
J
Yang
Department of Immunology, Neuroscience Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, State Key Laboratory of Medical Molecular Biology , Beijing, 100005, China.
Wei
W
He
Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine , Baltimore, MD 21205.
Hong
H
Wang
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Stephen M
0000-0002-9678-1192
SM
Eacker
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Zhikai
Z
Chi
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Xiaobo
0000-0001-6587-556X
X
Mao
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Jinchong
J
Xu
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Haisong
H
Jiang
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Shaida A
SA
Andrabi
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Ted M
0000-0002-6459-0893
TM
Dawson
Neuroregeneration Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205; Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Valina L
0000-0002-2915-3970
VL
Dawson
D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology; D016923:Cell Death / Q000187:drug effects / Q000502:physiology*; D015687:Cell Hypoxia / Q000502:physiology; D002478:Cells, Cultured; D019295:Computational Biology; D004195:Disease Models, Animal; D005947:Glucose / Q000172:deficiency; D020244:Infarction, Middle Cerebral Artery; D008297:Male; D008769:Methylnitronitrosoguanidine / Q000633:toxicity; D018345:Mice, Knockout; D009043:Motor Activity / Q000502:physiology; D058953:Neural Stem Cells / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D034622:RNA Interference; D054397:Receptors, CCR3 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D020521:Stroke / Q000378:metabolism / Q000473:pathology
C515351:Ccr3 protein, mouse; D054397:Receptors, CCR3; D008769:Methylnitronitrosoguanidine; D005947:Glucose
excitotoxicity; ischemia; library screen; stroke