Long non-coding RNAs (lncRNAs) have emerged as critical regulators in human disease including atherosclerosis. However, the mechanisms involved in the post-transcriptional regulation of the expression of disease-associated lncRNAs are not fully understood. Gene expression studies revealed that Nuclear Paraspeckle Assembly Transcript 1 (NEAT1) lncRNA expression was increased by >2-fold in peripheral blood mononuclear cells (PBMCs) derived from patients with coronary artery disease (CAD) or in carotid artery atherosclerotic plaques. We observed a linear association between NEAT1 lncRNA expression and prevalence of CAD which was independent of age, sex, cardiovascular traditional risk factors and renal function. NEAT1 expression was induced by TNF-α, while silencing of NEAT1 profoundly attenuated the TNF-α-induced vascular endothelial cell pro-inflammatory response as defined by the expression of CXCL8, CCL2, VCAM1 and ICAM1. Overexpression of the RNA editing enzyme adenosine deaminase acting on RNA-1 (ADAR1), but not of its editing-deficient mutant, upregulated NEAT1 levels. Conversely, silencing of ADAR1 suppressed the basal levels and the TNF-α-induced increase of NEAT1. NEAT1 lncRNA expression was strongly associated with ADAR1 in CAD and peripheral arterial vascular disease. RNA editing mapping studies revealed the presence of several inosines in close proximity to AU-rich elements within the AluSx3⁺/AluJo^‐ double-stranded RNA complex. Silencing of the stabilizing RNA-binding protein AUF1 reduced NEAT1 levels while silencing of ADAR1 profoundly affected the binding capacity of AUF1 to NEAT1. Together, our findings propose a mechanism by which ADAR1-catalyzed A-to-I RNA editing controls NEAT1 lncRNA stability in ASCVD.

长链非编码 RNA (lncRNA) 已成为包括动脉粥样硬化在内的人类疾病的关键调节因子。然而，疾病相关lncRNA表达的转录后调控机制尚不完全清楚。基因表达研究表明，来自冠状动脉疾病 (CAD) 患者或颈动脉粥样硬化斑块的外周血单个核细胞 (PBMC) 中，Nuclear Paraspeckle Assembly Transcript 1 ( NEAT1) lncRNA 表达增加了 2 倍以上。我们观察到NEAT1 lncRNA 表达与 CAD 患病率之间存在线性关联，该关联与年龄、性别、心血管传统危险因素和肾功能无关。整洁1表达由 TNF-α 诱导，而NEAT1的沉默显着减弱了 TNF-α 诱导的血管内皮细胞促炎反应，如CXCL8、CCL2、VCAM1和ICAM1的表达所定义的。作用于 RNA-1 (ADAR1) 的 RNA 编辑酶腺苷脱氨酶的过表达，但不是其编辑缺陷突变体的过表达，上调了 NEAT1水平。相反，ADAR1 的沉默抑制了基础水平和 TNF-α 诱导的NEAT1增加。整洁1lncRNA 表达与 CAD 和外周动脉血管疾病中的 ADAR1 密切相关。RNA 编辑作图研究表明，在AluSx3 ⁺ / AluJo ^-双链 RNA 复合物中，在富含 AU 的元素附近存在几种肌苷。稳定 RNA 结合蛋白 AUF1 的沉默降低了 NEAT1的水平，而 ADAR1 的沉默则深刻地影响了 AUF1 与NEAT1 的结合能力。总之，我们的研究结果提出了 ADAR1 催化的 A-to-I RNA 编辑控制ASCVD中 NEAT1 lncRNA 稳定性的机制。