Novel specialized pro-resolving lipid mediators in resolution of aortic aneurysms and rupture

项目总结 腹主动脉瘤(AAA)的形成和随后的主动脉破裂可导致猝死，是一种 严重的临床问题，目前尚无已知的治疗方法。我们最近的研究表明 具有专门的促分解介质(SPM)的保护作用，这些SPM是ω-3衍生的脂质 衍生物，即Resolvin(RV)D1，通过调节M1/M2有效地减弱AAA的形成 巨噬细胞极化。在这个提案中，我们将描述生物活性的表型和机制。 SPM的异构体，即Resolvins(Rvd1)，maresins(MAR-1)，Protectins(Pd-1)和Lipoxins(LxB4)， 可导致主动脉瘤形成的消退，防止主动脉破裂。首先，我们将在以下方面衡量SPM 人AAA患者和小鼠实验性AAA模型的电喷雾串联质量 LC/ESI-MS/MS联用是一种灵敏的分析方法 用于脂类介体的定性和定量分析。然后，我们将对协同作用进行表征 这些SPM生物活性亚型在我们的小鼠AAA和主动脉破裂模型中的反应。最后，我们会 描述SPM的生物活性亚型，即RvD1、MAR-1、PD-1和LxB4的具体机制 体内和体外研究。我们的初步数据表明，使用MAR-1治疗可以预防 与主动脉平滑肌细胞转化生长因子-β1信号转导相关的动脉瘤形成进展。近期 研究表明，氧化线粒体(Mt)dna作为损伤相关的模式分子形成。 篮网，但它仍然是AAA的特征。因此，MAR-1在促进巨噬细胞- 与Net相关的依赖性泡出(中性粒细胞凋亡/坏死性摄取的过程) 在人类AAA组织和实验小鼠模型中的形成也将被阐明。此外，我们 将对SPM相关的G蛋白偶联受体(GPCR)进行量化，即(ALX/FPR2，GPR32，GPR18和 ChemR23)AAA中的信号转导。具体地说，RvD1/FPR2通过AMP激活的蛋白激酶(AMPK)信号转导到 调节促炎巨噬细胞分泌GM-CSF和HMGB1将在AAA形成中破译。 因此，我们将描述SPM在治疗预先形成的动脉瘤和预防 通过调节RvD1-FPR2信号，线粒体DNA依赖的NETsis，促进主动脉破裂 泡腾作用和调节SMC依赖的转化生长因子-β-1信号。这一多方面的有针对性的方法 特殊的SPM亚型显示了一种临床适用的治疗慢性主动脉病变的策略 通过靶向巨噬细胞、中性粒细胞和平滑肌细胞的炎症和血管重塑。我们的 科学的前提是对这些促进分解的脂质衍生介质的作用进行深入分析 在动脉瘤形成的慢性模型中。重要的是，我们将测试它们的严密性和重现性 动物研究，以确定这些转变是否发生在人体组织中，使用体外主动脉移植。

PROJECT SUMMARY Abdominal aortic aneurysms (AAA) formation and subsequent aortic rupture can lead to sudden death and is a significant clinical problem with no currently known medical treatments available. Our recent studies have characterized a protective role of specialized pro-resolving mediators (SPMs) that are ω-3-derived lipid derivatives i.e. Resolvin (Rv)D1 that effectively attenuates AAA formation via modulating the M1/M2 macrophage polarization. In this proposal, we will delineate the phenotype and mechanisms of bioactive isoforms of SPMs i.e. Resolvins (RvD1), Maresins (MaR-1), Protectins (PD-1) and Lipoxins (LxB4) that can lead to resolution of aortic aneurysm formation and prevent aortic rupture. First, we will measure SPMs in human AAA patients and aortic tissue from murine experimental AAA model via electrospray tandem mass spectrometry coupled to liquid chromatography (LC/ESI-MS/MS), which is a sensitive analytical methodology for the qualitative and quantitative analysis of lipid mediators. Then, we will the characterize the synergistic response of these SPM bioactive isoforms in our murine AAA and aortic rupture models. Finally, we will delineate the specific mechanisms of the bioactive isoforms of SPMs i.e. RvD1, MaR-1, PD-1 and LxB4 using in vivo and in vitro studies. Our preliminary data demonstrates that treatment with MaR-1 can prevent the progression of aneurysm formation that is associated with aortic smooth muscle cell-TGF-β1 signaling. Recent reports suggest that oxidized mitochondrial (mt) DNA acts as a damage associated pattern molecule to form NETs, but it remains to be characterized in AAAs. Therefore, the role of MaR-1 in promoting macrophage- dependent efferocytosis (the process of uptake of apoptotic/necrotic neutrophils) associated with NET formation will also be elucidated in human AAA tissue as well as experimental murine models. Moreover, we will quantify SPM-related G-protein coupled receptors (GPCR) i.e. (ALX/FPR2, GPR32, GPR18, and ChemR23) signaling in AAA. Specifically, RvD1/FPR2 signaling via AMP-activated protein kinase (AMPK) to regulate pro-inflammatory macrophage secretion of GM-CSF and HMGB1 will be deciphered in AAA formation. Therefore, we will characterize the unique ability of SPMs to treat preformed aneurysms and prevent aortic rupture by modulating RvD1-FPR2 signaling, mtDNA-dependent NETosis, promoting efferocytosis and modulating SMC-dependent TGF-β1 signaling. This multi-faceted targeted approach of specific SPM isoforms demonstrate a clinically applicable therapeutic strategy for treatment of chronic aortic inflammation and vascular remodeling by targeting macrophages, neutrophils and smooth muscle cells. Our scientific premise is to perform an in-depth analysis of the roles of these pro-resolving lipid derived mediators in the chronic models of aneurysm formation. Importantly, we will test the rigor and reproducibility of these animal studies to determine whether these transitions occur in human tissue using ex vivo aortic explants.