LncRNA SNHG12, vascular senescence, and atherosclerosis

LncRNA SNHG12、血管衰老和动脉粥样硬化

基本信息

批准号：
10395512
负责人：
MARK W FEINBERG
金额：
$ 59.26万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-15 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10395512
关键词：
Arterial Fatty Streak Atherosclerosis Attention Bioinformatics Biological Assay Biological Process Biology Biophysics Blood Vessels Cause of Death Cell Adhesion Molecules Cell Aging Characteristics Chromatin Remodeling Factor Chronic Chronic Disease Code DNA Damage DNA Markers DNA-dependent protein kinase Deletion Mutation Disease Disease Progression Endothelial Cells Endothelium Family suidae Foundations Functional disorder Gamma-H2AX Genes Goals Human Hyperlipidemia Immunoprecipitation In Vitro Inflammation Inflammation Mediators Inflammatory Inflammatory Response Lesion Leukocytes Link Lipids Low-Density Lipoproteins Mass Spectrum Analysis Mediating Molecular Mus Myocardial Infarction Pathway interactions Peripheral Vascular Diseases Permeability Phase Play Process Proteins RNA Regulation Reporter Genes Repression Research Risk Factors Rodent Role SIRT1 gene Signal Pathway Signal Transduction Small Nucleolar RNA Stimulus Stroke TP53 gene Therapeutic Transgenic Mice Untranslated RNA Vascular Cell Adhesion Molecule-1 Vascular Diseases Vascular Endothelium atherogenesis base cell type chemokine cytokine frontier in vivo insight molecular imaging multidisciplinary nanomedicine nanoparticle nicotinamide-beta-riboside novel novel therapeutic intervention recruit response senescence transcriptome sequencing transcytosis

项目摘要

Long non-coding RNAs (lncRNAs) have garnered widespread attention as emerging regulators of diverse biological processes relevant to atherosclerosis. However, the identity and roles of specific lncRNAs within atherosclerotic lesions are not well defined. Using RNA-Seq profiling to identify lncRNAs derived specifically from the aortic intima of LDLR-/- mice during lesion progression and regression phases, we identify the lncRNA Small Nucleolar Host Gene-12 (SNHG12). SNHG12 is highly enriched in the vascular endothelium across mice, pigs, and humans and is significantly reduced with atherosclerotic lesion progression, but increased with regression. Our preliminary studies show that gapmeR-mediated silencing of SNHG12 potently accelerated atherosclerotic lesion formation by over 2-fold in LDLR-/- mice. Remarkably, the increased lesional effects were not driven by lipid-lowering or by inflammatory recruitment of lesional leukocytes, but rather by increased DNA damage (γH2AX) and senescence (p16, p21, p27) in the vascular endothelium. Accumulating studies demonstrate that vascular senescence induced by the DNA damage response (DDR) may adversely contribute to chronic inflammation in atherosclerotic lesions. However, the mechanisms linking senescence and atherosclerotic lesion formation remain poorly understood. LncRNAs play important regulatory roles by interacting with RNA, chromatin modifiers, or protein- coding genes. Mechanistically, using a modified RNA IP (RIP)-mass spectrometry pulldown assay, we identify that SNHG12 interacts with the DNA-dependent protein kinase (DNA-PK) to control the DNA-damage response. Preliminary studies show SNHG12 deficiency in ECs significantly increased DNA damage, markers of senescence, and EC permeability to LDL. Moreover, we demonstrate that the NAD+ precursor nicotinamide riboside (NR), that suppresses endothelial senescence, may function in an SNHG12-dependent manner. These observations provide the foundation for the central hypothesis that endothelial SNHG12 deficiency, via regulatory effects on DNA-PK and the DNA damage response, promotes vascular senescence, senescence- associated inflammation, and atherosclerosis. To elucidate this further, three aims are proposed. In Aim1, we will delineate the molecular basis for SNHG12's ability to regulate DNA-PK-mediated DNA damage response and vascular senescence in ECs. In Aim2, we will determine the effect of altering lncRNA SNHG12 expression in an EC-specific manner on the DNA damage response and atherosclerotic progression and regression. In Aim3, we will explore the molecular mechanisms by which stimuli repress and NR rescues SNHG12 expression in ECs, and we will determine whether the anti-senescent effects of NR are SNHG12-dependent. This multi-disciplinary team in the fields of non-coding RNA biology, molecular imaging, nanomedicine, bioinformatics, and atherosclerosis research will establish an unprecedented molecular view of this lncRNA in lesions that can inform a new frontier in the regulation of vascular senescence and atherosclerosis.

长链非编码RNA（lncRNA）作为多种生物多样性的新兴调节剂而受到广泛关注。然而，与动脉粥样硬化相关的生物过程中特定lncRNA的身份和作用。动脉粥样硬化病变尚未明确定义，但使用 RNA-Seq 分析来识别特异性衍生的 lncRNA。从 LDLR-/- 小鼠在病变进展和消退阶段的主动脉内膜中，我们鉴定了 lncRNA 小核仁宿主基因 12 (SNHG12) 在血管内皮细胞中高度富集。小鼠、猪和人类，随着动脉粥样硬化病变进展而显着减少，但随着动脉粥样硬化病变进展而增加我们的初步研究表明，gapmeR 介导的 SNHG12 沉默有效加速。 LDLR-/- 小鼠中动脉粥样硬化病变的形成增加了 2 倍以上。不是由降脂或病变白细胞的炎症募集驱动，而是由 DNA 增加驱动血管内皮损伤（γH2AX）和衰老（p16、p21、p27）的积累研究。证明 DNA 损伤反应 (DDR) 诱导的血管衰老可能会产生不利影响然而，与衰老和动脉粥样硬化病变相关的机制。动脉粥样硬化病变的形成仍然知之甚少。 LncRNA 通过与 RNA、染色质修饰剂或蛋白质相互作用发挥重要的调节作用从机制上讲，我们使用改良的 RNA IP (RIP) 质谱下拉分析来识别。 SNHG12 与 DNA 依赖性蛋白激酶 (DNA-PK) 相互作用以控制 DNA 损伤初步研究表明，EC 中的 SNHG12 缺陷显着增加了 DNA 损伤、标记物。此外，我们还证明了 NAD+ 前体烟酰胺的作用。抑制内皮衰老的核糖苷 (NR) 可能以 SNHG12 依赖性方式发挥作用。这些观察结果为内皮细胞 SNHG12 缺乏通过对DNA-PK和DNA损伤反应的调节作用，促进血管衰老，衰老- 为了进一步阐明这一点，我们在 Aim1 中提出了三个目标。将描绘 SNHG12 调节 DNA-PK 介导的 DNA 损伤反应能力的分子基础在 Aim2 中，我们将确定改变 lncRNA SNHG12 表达的影响。以 EC 特有的方式影响 DNA 损伤反应以及动脉粥样硬化的进展和消退。 Aim3，我们将探索刺激抑制和NR拯救SNHG12的分子机制 ECs 中的表达，我们将确定 NR 的抗衰老作用是否依赖于 SNHG12。这个多学科团队在非编码RNA生物学、分子成像、纳米医学、生物信息学和动脉粥样硬化研究将为该 lncRNA 建立前所未有的分子观点病变可以为血管衰老和动脉粥样硬化的调节提供新的前沿。