Neutrophil extracellular traps promote tPA-induced brain hemorrhage via cGAS in mice with stroke
Ranran Wang, Yuanbo Zhu, Zhongwang Liu, Luping Chang, Xiaofei Bai, Lijing Kang, Yongliang Cao, Xing Yang, Huilin Yu, Mei-Juan Shi, Yue Hu, Wenying Fan, Bing-Qiao Zhao, Ranran Wang, Yuanbo Zhu, Zhongwang Liu, Luping Chang, Xiaofei Bai, Lijing Kang, Yongliang Cao, Xing Yang, Huilin Yu, Mei-Juan Shi, Yue Hu, Wenying Fan, Bing-Qiao Zhao
Abstract
Intracerebral hemorrhage associated with thrombolytic therapy with tissue plasminogen activator (tPA) in acute ischemic stroke continues to present a major clinical problem. Here, we report that infusion of tPA resulted in a significant increase in markers of neutrophil extracellular traps (NETs) in the ischemic cortex and plasma of mice subjected to photothrombotic middle cerebral artery occlusion. Peptidylarginine deiminase 4 (PAD4), a critical enzyme for NET formation, is also significantly upregulated in the ischemic brains of tPA-treated mice. Blood-brain barrier (BBB) disruption after ischemic challenge in an in vitro model of BBB was exacerbated after exposure to NETs. Importantly, disruption of NETs by DNase I or inhibition of NET production by PAD4 deficiency restored tPA-induced loss of BBB integrity and consequently decreased tPA-associated brain hemorrhage after ischemic stroke. Furthermore, either DNase I or PAD4 deficiency reversed tPA-mediated upregulation of the DNA sensor cyclic GMP-AMP (cGAMP) synthase (cGAS). Administration of cGAMP after stroke abolished DNase I-mediated downregulation of the STING pathway and type 1 interferon production and blocked the antihemorrhagic effect of DNase I in tPA-treated mice. We also show that tPA-associated brain hemorrhage after ischemic stroke was significantly reduced in cGas-/- mice. Collectively, these findings demonstrate that NETs significantly contribute to tPA-induced BBB breakdown in the ischemic brain and suggest that targeting NETs or cGAS may ameliorate thrombolytic therapy for ischemic stroke by reducing tPA-associated hemorrhage.
Conflict of interest statement
Conflict-of-interest disclosure: The authors declare no competing financial interests.
© 2021 by The American Society of Hematology.
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References
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