Diabetes Mellitus-Induced Long Noncoding RNA Dnm3os Regulates Macrophage Functions and Inflammation via Nuclear Mechanisms.
Diabetes Mellitus-Induced Long Noncoding RNA Dnm3os Regulates Macrophage Functions and Inflammation via Nuclear Mechanisms.
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DOI：
10.1161/atvbaha.117.310663
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发表时间：
2018-08
期刊：
影响因子：
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通讯作者：
Natarajan R
中科院分区：
文献类型：
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作者：
Das S;Reddy MA;Senapati P;Stapleton K;Lanting L;Wang M;Amaram V;Ganguly R;Zhang L;Devaraj S;Schones DE;Natarajan R
巨噬细胞在炎症和糖尿病血管并发症中起关键作用。新出现的证据表明，长非编码RNA(LncRNAs)与炎症有关，但它们在与炎症性糖尿病并发症相关的巨噬细胞功能障碍中的作用尚不清楚，因此本研究对其进行了研究。Rna测序和RT-qPCR结果表明，与对照组相比，2型糖尿病db/db小鼠、饮食诱导的胰岛素抵抗小鼠和糖尿病载脂蛋白E−/−小鼠的骨髓来源的巨噬细胞以及2型糖尿病患者的单核细胞中都上调了lncRNA Dynamin 3反向链(Dnm3os)。糖尿病条件(高糖和棕榈酸)诱导小鼠和人巨噬细胞中的Dnm3os。启动子报告分析和染色质免疫沉淀分析表明，糖尿病条件下通过激活NF-κB诱导Dnm3os。亚细胞部分的RNA-FISH和RT-qPCRs显示巨噬细胞中Dnm3os的核定位和染色质丰富。巨噬细胞中Dnm3os的稳定过表达改变了组蛋白的整体修饰，上调了炎症和免疫反应基因，以及吞噬功能。相反，RNAi介导的Dnm3os基因敲除减弱了这些反应。巨噬细胞核裂解产物的RNA下拉实验证实核仁素和ILF-2为Dnm3os的蛋白结合伙伴，并经RNA-IP和RNA-FISH免疫荧光进一步证实。此外，在糖尿病情况下，核仁素水平降低，其基因敲除增强了Dnm3os诱导的炎症基因表达及其启动子上的组蛋白H3K9乙酰化。这些结果显示了新的机制，涉及LncRNA Dnm3os的上调，它与核仁素的相互作用中断，以及促进糖尿病巨噬细胞炎症表型的靶基因的表观遗传修饰。这些数据可能导致基于lncRNA的炎症性糖尿病并发症的治疗。
Macrophages play key roles in inflammation and diabetic vascular complications. Emerging evidence implicates long noncoding RNAs (lncRNAs) in inflammation, but their role in macrophage dysfunction associated with inflammatory diabetic complications is unclear and was therefore investigated in this study. RNA-sequencing and RT-qPCR demonstrated that a lncRNA Dynamin 3 opposite strand (Dnm3os) is upregulated in bone marrow derived macrophages from type 2 diabetic db/db mice, diet-induced insulin-resistant mice and diabetic ApoE−/− mice, as well as in monocytes from type 2 diabetic patients relative to controls. Diabetic conditions (High glucose and palmitic acid) induced Dnm3os in mouse and human macrophages. Promoter reporter analysis and chromatin-immunoprecipitation assays demonstrated that diabetic conditions induce Dnm3os via NF-κB activation. RNA-FISH and RT-qPCRs of sub-cellular fractions demonstrated nuclear localization and chromatin enrichment of Dnm3os in macrophages. Stable overexpression of Dnm3os in macrophages altered global histone modifications and upregulated inflammation and immune response genes, and phagocytosis. Conversely, RNAi-mediated knockdown of Dnm3os attenuated these responses. RNA-pull-down assays with macrophage nuclear lysates identified nucleolin and ILF-2 as protein binding partners of Dnm3os, which was further confirmed by RNA-IP and RNA-FISH-immunofluorescence. Furthermore, nucleolin levels were decreased in diabetic conditions, and its knockdown enhanced Dnm3os-induced inflammatory gene expression and histone H3K9-acetylation at their promoters. These results demonstrate novel mechanisms involving upregulation of lncRNA Dnm3os, disruption of its interaction with nucleolin, and epigenetic modifications at target genes that promote macrophage inflammatory phenotype in diabetes. The data could lead to lncRNA-based therapies for inflammatory diabetes complications.