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.

长期非编码RNA（LNCRNA）作为潜水员的新兴调节剂引起了广泛关注与动脉粥样硬化有关的生物过程。但是，特定lncrnas的身份和作用动脉粥样硬化病变的定义不当。使用RNA-seq分析来识别专门衍生的LNCRNA 在病变进展和回归阶段的LDLR - / - 小鼠的主动脉内膜中，我们识别LNCRNA 小核仁宿主基因12（SNHG12）。 SNHG12高度富集在横跨的血管内皮中小鼠，猪和人类，随着动脉粥样硬化病变的进展显着降低，但随着增加回归。我们的初步研究表明，Gapmer介导的SNHG12的沉默可能加速了在LDLR - / - 小鼠中，动脉粥样硬化病变形成超过2倍。值得注意的是，增长的病变影响是不是由脂质降低或受炎症性白细胞募集的驱动，而是通过增加的DNA驱动血管内皮中的损伤（γH2AX）和敏感（p16，p21，p27）。累积研究证明由DNA损伤响应（DDR）诱导的血管感应可能会造成不利影响在动脉粥样硬化病变中慢性炎症。但是，连接感应和的机制动脉粥样硬化病变的形成仍然很少了解。 LNCRNA通过与RNA，染色质修饰剂或蛋白质 - 编码基因。从机械上讲，使用改良的RNA IP（RIP） - 质谱式下拉测定法，我们确定 SNHG12与DNA依赖性蛋白激酶（DNA-PK）相互作用以控制DNA破坏回复。初步研究表明，EC的SNHG12缺乏显着增加了DNA损伤，标记感应的，以及对LDL的渗透性。此外，我们证明了NAD+前体烟酰胺抑制内皮感应的核糖剂（NR）可能以SNHG12依赖性方式起作用。这些观察结果为内皮SNHG12缺乏症的中心假设奠定了基础调节对DNA-PK和DNA损伤反应的影响，促进血管感应，感应 - 相关的炎症和动脉粥样硬化。为了进一步阐明这一点，提出了三个目标。在AIM1中，我们将描绘SNHG12调节DNA-PK介导的DNA损伤响应的能力的分子基础 EC中的血管感应。在AIM2中，我们将确定改变LNCRNA SNHG12表达的效果以EC特异性方式对DNA损伤反应以及动脉粥样硬化的进展和回归。 AIM3，我们将探索刺激反射和NR拯救SNHG12的分子机制在EC中的表达，我们将确定NR的抗阳性效应是否依赖于SNHG12。这个多学科的团队在非编码RNA生物学，分子成像，纳米医学的领域生物信息学和动脉粥样硬化研究将在该lncRNA中建立前所未有的分子视图可以在调节血管感染和动脉粥样硬化的调节方面为新的边界提供信息的病变。