Data_Sheet_2_Dynamic A-to-I RNA editing during acute neuroinflammation in sepsis-associated encephalopathy.PDF

IntroductionThe activation of cerebral endothelial cells (CECs) has recently been reported to be the earliest acute neuroinflammation event in the CNS during sepsis-associated encephalopathy (SAE). Importantly, adenosine-to-inosine (A-to-I) RNA editing mediated by ADARs has been associated with SAE, yet its role in acute neuroinflammation in SAE remains unclear.
MethodsOur current study systematically analyzed A-to-I RNA editing in cerebral vessels, cerebral endothelial cells (CECs), and microglia sampled during acute neuroinflammation after treatment in a lipopolysaccharide (LPS)-induced SAE mouse model.
ResultsOur results showed dynamic A-to-I RNA editing activity changes in cerebral vessels during acute neuroinflammation. Differential A-to-I RNA editing (DRE) associated with acute neuroinflammation were identified in these tissue or cells, especially missense editing events such as S367G in antizyme inhibitor 1 (Azin1) and editing events in lincRNAs such as maternally expressed gene 3 (Meg3), AW112010, and macrophage M2 polarization regulator (Mm2pr). Importantly, geranylgeranyl diphosphate synthase 1 (Ggps1) and another three genes were differentially edited across cerebral vessels, CECs, and microglia. Notably, Spearman correlation analysis also revealed dramatic time-dependent DRE during acute neuroinflammation, especially in GTP cyclohydrolase1 (Gch1) and non-coding RNA activated by DNA damage (Norad), both with the editing level positively correlated with both post-LPS treatment time and edited gene expression in cerebral vessels and CECs.
DiscussionThe findings in our current study demonstrate substantial A-to-I RNA editing changes during acute neuroinflammation in SAE, underlining its potential role in the disease.

**引言** 目前已有研究表明，在脓毒症相关性脑病（sepsis-associated encephalopathy, SAE）过程中，脑内皮细胞（cerebral endothelial cells, CECs）的激活是中枢神经系统（central nervous system, CNS）内最早出现的急性神经炎症事件。值得注意的是，由ADARs介导的腺苷肌苷（adenosine-to-inosine, A-to-I）RNA编辑已被证实与SAE相关，但其在SAE急性神经炎症中的作用仍未明确。**方法** 本研究通过脂多糖（lipopolysaccharide, LPS）诱导构建SAE小鼠模型，对造模后急性神经炎症阶段采集的脑血管、脑内皮细胞（CECs）及小胶质细胞样本中的A-to-I RNA编辑情况进行了系统性分析。**结果** 本研究结果显示，急性神经炎症过程中脑血管内的A-to-I RNA编辑活性呈现动态变化。在上述组织及细胞中，我们鉴定出了与急性神经炎症相关的差异A-to-I RNA编辑（differential A-to-I RNA editing, DRE）事件，尤其是抗酶抑制剂1（antizyme inhibitor 1, Azin1）中S367G这类错义编辑事件，以及母系表达基因3（maternally expressed gene 3, Meg3）、AW112010、巨噬细胞M2极化调节因子（macrophage M2 polarization regulator, Mm2pr）等长链基因间非编码RNA（long intergenic non-coding RNAs, lincRNAs）中的编辑事件。值得注意的是，牻牛儿基牻牛儿基二磷酸合酶1（geranylgeranyl diphosphate synthase 1, Ggps1）及另外3个基因在脑血管、CECs与小胶质细胞中均存在差异编辑现象。此外，斯皮尔曼相关分析（Spearman correlation analysis）还揭示了急性神经炎症过程中显著的时间依赖性DRE，尤其在GTP环水解酶1（GTP cyclohydrolase1, Gch1）和DNA损伤激活的非编码RNA（non-coding RNA activated by DNA damage, Norad）中：这两种基因的编辑水平在脑血管及CECs中，均与脂多糖处理后的时间及编辑基因的表达量呈正相关。**讨论** 本研究结果证实，SAE相关急性神经炎症过程中存在广泛的A-to-I RNA编辑变化，凸显了其在该疾病中潜在的调控作用。