Whole-Transcriptome Sequencing Analyses of Nuclear Antixoxidant-1 in Endothelial Cells: Role in Inflammation and Atherosclerosis.
Whole-Transcriptome Sequencing Analyses of Nuclear Antixoxidant-1 in Endothelial Cells: Role in Inflammation and Atherosclerosis.
复制标题
DOI：
10.3390/cells11182919
复制
发表时间：
2022-09-18
期刊：
影响因子：
6
通讯作者：
Fukai, Tohru
中科院分区：
文献类型：
--
作者：
Sudhahar, Varadarajan;Shi, Yang;Kaplan, Jack H.;Ushio-Fukai, Masuko;Fukai, Tohru
炎症、氧化应激和铜（Cu）在心血管疾病（包括动脉粥样硬化）中起重要作用。我们之前报道了细胞质Cu伴侣抗氧化剂-1 （Atox1）在炎症细胞因子或外源性Cu的作用下易位到细胞核，并且Atox1定位于炎症动脉粥样硬化主动脉内皮的细胞核。然而，人们对核Atox1的作用及其功能知之甚少。在这里，我们发现ApoE−/−小鼠的Atox1缺乏与西方饮食显著减少动脉粥样硬化病变的形成。体外，在体外培养的人内皮细胞（ECs）中，腺病毒介导的核靶向Atox1 （Ad-Atox1-NLS）的过表达比对照细胞（Ad-null）增加了单核细胞粘附和活性氧（ROS）的产生。为了解决潜在的机制，我们使用无偏倚的系统方法整合测序数据，对ECs中atox1调控靶点进行了全基因组定位。结合ChIP-Seq和RNA-Seq对转染Ad-Atox1-NLS或Ad-null的ECs进行分析，鉴定出1387个差异表达基因（DEG）。Motif富集实验和KEGG通路富集分析显示，Atox1-NLS调控了248个差异表达基因，包括炎症和血管生成基因，并通过实时荧光定量pcr证实了这一结果。在这些基因中，炎症反应的功能分析发现CD137、CSF1和IL5RA是新的核atox1靶向炎症基因，而CD137也是atox1 - nls诱导的ROS产生的关键调节因子。这些发现揭示了参与炎症和ROS产生的新的核Atox1下游靶点，并为核Atox1作为治疗炎症性疾病（如动脉粥样硬化）的潜在治疗靶点提供了见解。
Inflammation, oxidative stress, and copper (Cu) play an important role in cardiovascular disease, including atherosclerosis. We previously reported that cytosolic Cu chaperone antioxidant-1 (Atox1) translocates to the nucleus in response to inflammatory cytokines or exogenous Cu and that Atox1 is localized at the nucleus in the endothelium of inflamed atherosclerotic aorta. However, the roles of nuclear Atox1 and their function are poorly understood. Here we showed that Atox1 deficiency in ApoE−/− mice with a Western diet exhibited a significant reduction of atherosclerotic lesion formation. In vitro, adenovirus-mediated overexpression of nuclear-targeted Atox1 (Ad-Atox1-NLS) in cultured human endothelial cells (ECs) increased monocyte adhesion and reactive oxygen species (ROS) production compared to control cells (Ad-null). To address the underlying mechanisms, we performed genome-wide mapping of Atox1-regulated targets in ECs, using an unbiased systemic approach integrating sequencing data. Combination of ChIP-Seq and RNA-Seq analyses in ECs transfected with Ad-Atox1-NLS or Ad-null identified 1387 differentially expressed genes (DEG). Motif enrichment assay and KEGG pathway enrichment analysis revealed that 248 differentially expressed genes, including inflammatory and angiogenic genes, were regulated by Atox1-NLS, which was then confirmed by real-time qPCR. Among these genes, functional analysis of inflammatory responses identified CD137, CSF1, and IL5RA as new nuclear Atox1-targeted inflammatory genes, while CD137 is also a key regulator of Atox1-NLS-induced ROS production. These findings uncover new nuclear Atox1 downstream targets involved in inflammation and ROS production and provide insights into the nuclear Atox1 as a potential therapeutic target for the treatment of inflammatory diseases such as atherosclerosis.
登录
查看更多内容
影响因子：
20.1
作者：
Jeney, V;Itoh, S;Fukai, T
通讯作者：
Fukai, T
影响因子：
4.8
作者：
Itoh, Shinichi;Kim, Ha Won;Fukai, Tohru
通讯作者：
Fukai, Tohru
影响因子：
3
作者：
Beaino, Wissam;Guo, Yunjun;Anderson, Carolyn J.
通讯作者：
Anderson, Carolyn J.
影响因子：
9.7
作者：
Jin, Jing;Ma, Mingjun;Teng, Chun-Bo
通讯作者：
Teng, Chun-Bo
影响因子：
4.4
作者：
Gregory, B;Kirchem, A;Robinson, DS
通讯作者：
Robinson, DS