Macrophage small RNA export to HDL protects against inflammation and atherosclerosis

巨噬细胞向 HDL 输出小 RNA 可预防炎症和动脉粥样硬化

基本信息

批准号：
10089342
负责人：
Kasey C Vickers
金额：
$ 38.93万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10089342
关键词：
Agonist Apolipoprotein E Area Arterial Fatty Streak Atherosclerosis Attenuated Binding Biological Process Blood Circulation Cardiometabolic Disease Cardiovascular Diseases Cells Chronic Complex Deposition Disease Due Process Endocytosis Environment Functional disorder Gene Expression High Density Lipoproteins Human Immune Impairment In Vitro Inflammation Inflammatory Knowledge Lesion Lipids Lipoproteins Literature Low-Density Lipoproteins Lysosomes Macrophage Activation Mediating MicroRNAs Modeling Modification Mus Pathology Pathway interactions Phagocytosis Phenotype Process Property Receptor Activation Reporting Route Signal Transduction Small RNA Stimulus TLR7 gene TLR8 gene Testing Therapeutic Toll-like receptors Untranslated RNA Work base cytokine extracellular field study human disease in vivo inhibitor/antagonist lipid metabolism locked nucleic acid macrophage microbial microbiome mouse model novel nucleic acid inhibitor particle receptor response single-cell RNA sequencing therapy design uptake western diet

项目摘要

Summary: Atherosclerosis is the underlying cause for ischemic cardiovascular disease (CVD), a pathology best characterized as uncontrolled inflammation in response to continual lipid deposition in the arterial wall. Despite the importance of inflammation in the progression of atherosclerosis, very few therapies are designed to target this process due to a general lack of knowledge of atherogenic stimuli outside of bioactive lipids. Previously, we have reported that lipoproteins, namely HDL and LDL, transport small non-coding RNAs (sRNA), and recently, we discovered that lipoproteins are highly-enriched with sRNAs derived from bacterial and fungal species in the microbiome and environment. Interestingly, these foreign, microbial sRNAs (msRNAs) are also highly-abundant in macrophages within the atherosclerotic lesions and activate sRNA-sensing toll-like receptors (TLR)7/8 in lesion macrophages. We posit that HDL accepts msRNAs from lesion macrophages, and thus, suppresses TLR7/8 activation and down-stream pro-inflammatory gene expression. Supporting this model, macrophages were found to readily export msRNAs to HDL and HDL treatments were found to block msRNA-induced activation. In CVD, HDL acquire reactive dicarbonyl modifications which are associated with HDL dysfunction. In preliminary studies, we found that reactive dicarbonyl modifications on HDL, e.g. isolevuglandins (IsoLG), impair HDL-msRNA export. Moreover, we found that IsoLG-modified HDL had increased macrophage uptake and retention and induced pro-inflammatory gene expression. Preliminary studies also found that treating mouse models with reactive scavengers to block IsoLG modifications decreased atherosclerosis. Therefore, we aimed to block msRNA activation of TLR7/8 using non-targeting locked-nucleic acids (ntLNA). Strikingly, we found that ntLNA treatments significantly reduced atherosclerotic lesion area by 30% over 4 weeks in Apoe-/- mice on western diet. In addition, single-cell RNA sequencing approaches demonstrated that ntLNA therapy repressed pro-inflammatory gene expression in lesion macrophages and reduced the number of pro-inflammatory macrophages in atherosclerotic lesions. Based on preliminary studies, we hypothesize HDL removes msRNA from lesion macrophages through retro-endocytosis, a process that is inhibited by IsoLG modifications on HDL. Furthermore, we posit that msRNA activation of endo-lysosome TLR7/8 in lesion macrophages can be readily targeted with ntLNAs to reduce atherosclerosis. To test this hypothesis, we aim to I.) Determine the mechanism(s) and consequences of macrophage msRNA export to HDL, the impact of HDL-msRNA export on macrophage activation, and define the HDL-mediated reverse sRNA transport (RsRT) pathway in vivo, II.) Define the impact of dicarbonyl modifications and msRNA cargo on HDL dysfunction in CVD, and III.) Target macrophage msRNA receptors to inhibit atherosclerosis progression and promote regression. This project will open entirely new fields of study for lipoproteins, extracellular sRNAs, host/non-host interactions, and inflammation, which have applicability to many other diseases, but is particularly critical for CVD.

摘要：动脉粥样硬化是缺血性心血管疾病（CVD）的根本原因，这是一种最好的病理学响应于动脉壁中的持续脂质沉积而被表征为不受控制的炎症。尽管炎症在动脉粥样硬化进展中的重要性，很少有人设计用于靶向的疗法由于普遍缺乏对生物活性脂质之外的动脉粥样硬化刺激的知识，因此此过程。以前，我们据报道，脂蛋白，即HDL和LDL，转运小型非编码RNA（SRNA），最近，最近，我们发现，脂蛋白富含来自细菌和真菌种类的SRNA 微生物组和环境。有趣的是，这些外国微生物SRNA（MSRNA）也很丰富在动脉粥样硬化病变内的巨噬细胞中，并激活SRNA感应Toll样受体（TLR）7/8 病变巨噬细胞。我们认为HDL接受病变巨噬细胞中的MSRNA，从而抑制 TLR7/8激活和下游促炎基因表达。支持此模型，巨噬细胞发现很容易将MSRNA导出到HDL，并发现HDL处理可阻止MSRNA诱导的激活。在CVD中，HDL获得与HDL功能障碍相关的反应性二骨修饰。在初步研究中，我们发现对HDL的反应性双骨修改，例如Isolevuglandins（Isalg），损害HDL-MSRNA出口。此外，我们发现Isalg修饰的HDL的巨噬细胞吸收增加了保留和诱导促炎基因表达。初步研究还发现治疗小鼠具有反应性清除剂的模型阻止分离的修饰减少了动脉粥样硬化。因此，我们的目标使用非靶向锁定核酸（NTLNA）阻止TLR7/8的毫秒激活。令人惊讶的是，我们发现 NTLNA处理在APOE-/ - 小鼠中，在4周内将动脉粥样硬化病变区域显着降低了30％西方饮食。此外，单细胞RNA测序方法表明NTLNA治疗受到抑制病变巨噬细胞中的促炎基因表达，减少了促炎的数量动脉粥样硬化病变中的巨噬细胞。根据初步研究，我们假设HDL去除MSRNA 从病变巨噬细胞到恢复性细胞增多症，这一过程受到HDL的疾病修饰的抑制。此外，我们认为在病变巨噬细胞中内散糖体TLR7/8激活很容易用NTLNA靶向减少动脉粥样硬化。为了检验这一假设，我们的目标是I。）确定机理和巨噬细胞MSRNA导出到HDL的后果，HDL-MSRNA导出对HDL的影响巨噬细胞激活并定义了体内的HDL介导的反向SRNA转运（RSRT）途径，ii。）定义 Dicarbonyl修饰和MSRNA货物对CVD中HDL功能障碍的影响，以及III。巨噬细胞msRNA受体抑制动脉粥样硬化进展并促进消退。这个项目将开放脂蛋白，细胞外SRNA，宿主/非宿主相互作用的全新研究领域炎症，对许多其他疾病具有适用性，但对于CVD尤其重要。