Cardiology Division, Department of Medicine, Columbia University Irving Medical Center, New York, NY (E.C., M.E.O., A.C.B., H.Z., L.R., C.X., M.P.R.).
Mission Bio, South San Francisco, CA (D.Y.L.).
Arterioscler Thromb Vasc Biol. 2023 Feb;43(2):286-299. doi: 10.1161/ATVBAHA.122.318353. Epub 2022 Dec 22.
Long noncoding RNAs (lncRNAs) have emerged as novel regulators of macrophage biology and inflammatory cardiovascular diseases. However, studies focused on lncRNAs in human macrophage subtypes, particularly human lncRNAs that are not conserved in rodents, are limited.
Through RNA-sequencing of human monocyte-derived macrophages, we identified suppressor of inflammatory macrophage apoptosis lncRNA (). Lipopolysaccharide/IFNγ (interferon γ) stimulated human macrophages were treated with antisense oligonucleotides and subjected to RNA-sequencing to investigate the function of . Western blots, luciferase assay, and RNA immunoprecipitation were performed to validate function and potential mechanism of RNAscope was performed to identify expression in human carotid atherosclerotic plaques.
RNA-sequencing of human monocyte-derived macrophages identified , a human macrophage-specific long intergenic noncoding RNA that is highly induced in lipopolysaccharide/IFNγ-stimulated macrophages. knockdown in lipopolysaccharide/IFNγ stimulated THP1 human macrophages induced apoptosis of inflammatory macrophages, as shown by increased protein expression of cleaved PARP (poly[ADP-ribose] polymerase), caspase 9, caspase 3, and Annexin V+. RNA-sequencing of control versus knockdown in lipopolysaccharide/IFNγ-stimulated macrophages showed Netrin-1 () to be significantly decreased upon knockdown. We confirmed that knockdown in lipopolysaccharide/IFNγ-stimulated macrophages induced apoptosis. The knockdown-induced apoptotic phenotype was rescued by adding recombinant NTN1. promoter-luciferase reporter activity was increased in HEK293T (human embryonic kidney 293) cells treated with lentiviral overexpression of . promoter activity is known to require HIF1α (hypoxia-inducible factor 1 subunit alpha), and our studies suggest that may interact with HIF1α to regulate transcription, thereby regulating macrophages apoptosis. was found to be expressed in macrophages in human carotid atherosclerotic plaques of symptomatic patients.
is a nonconserved, human macrophage lncRNA expressed in atherosclerosis that suppresses macrophage apoptosis. partners with HIF1α (hypoxia-inducible factor 1 subunit alpha) to regulate NTN1, which is a known macrophage survival factor. This work illustrates the importance of interrogating the functions of human lncRNAs and exploring their translational and therapeutic potential in human atherosclerosis.
长非编码 RNA（lncRNA）已成为调节巨噬细胞生物学和炎症性心血管疾病的新调控因子。然而，目前针对人类巨噬细胞亚型中的 lncRNA 的研究还很有限，尤其是针对人类而非啮齿动物中保守的 lncRNA 的研究。
通过对人单核细胞来源的巨噬细胞进行 RNA 测序，我们鉴定出了抑制炎症性巨噬细胞凋亡 lncRNA（）。用反义寡核苷酸处理脂多糖/IFNγ（干扰素γ）刺激的人巨噬细胞，并进行 RNA 测序，以研究的功能。通过 Western blot、荧光素酶测定和 RNA 免疫沉淀实验来验证的功能和潜在机制。通过 RNAscope 检测人类颈动脉粥样硬化斑块中 lncRNA 的表达。
对人单核细胞来源的巨噬细胞进行 RNA 测序，鉴定出了一个高度诱导脂多糖/IFNγ刺激的巨噬细胞表达的人类巨噬细胞特异性长基因间非编码 RNA。在脂多糖/IFNγ 刺激的 THP1 人巨噬细胞中敲低后，诱导炎症性巨噬细胞凋亡，表现为多聚（ADP-核糖）聚合酶（PARP）、半胱氨酸天冬氨酸蛋白酶 9（caspase 9）、半胱氨酸天冬氨酸蛋白酶 3（caspase 3）和膜联蛋白 V 的蛋白表达增加。在脂多糖/IFNγ 刺激的巨噬细胞中进行对照与敲低的 RNA 测序显示，在敲低后 Netrin-1（）显著减少。我们证实了在脂多糖/IFNγ 刺激的巨噬细胞中敲低诱导了细胞凋亡。用重组 NTN1 处理可挽救敲低诱导的凋亡表型。在接受慢病毒过表达的 HEK293T（人胚肾 293）细胞中，的启动子-荧光素酶报告活性增加。已知的启动子活性需要 HIF1α（缺氧诱导因子 1 亚单位α），我们的研究表明可能与 HIF1α 相互作用以调节转录，从而调节巨噬细胞凋亡。在有症状患者的人类颈动脉粥样硬化斑块中的巨噬细胞中检测到。
是一种在动脉粥样硬化中表达的非保守人类巨噬细胞 lncRNA，可抑制巨噬细胞凋亡。与 HIF1α（缺氧诱导因子 1 亚单位α）合作调节 NTN1，这是一种已知的巨噬细胞存活因子。这项工作说明了研究人类 lncRNA 的功能以及探索其在人类动脉粥样硬化中的转化和治疗潜力的重要性。
