LysoPI/GPR55 pathway promotes endothelial activation, vascular inflammation and atherosclerosis

标题：LysoPI/GPR 55通路促进内皮活化、血管炎症和 粥样硬化 动脉粥样硬化及其并发症是美国发病率和死亡率的主要原因。小说 迫切需要抗动脉粥样硬化疗法来抑制动脉粥样硬化。该项目的目标是 确定溶血磷脂酰肌醇（lysoPI）促进内皮细胞增殖的作用和机制。 细胞（EC）激活和动脉粥样硬化。我们发表了许多关于EC激活的论文， 动脉粥样硬化和溶血磷脂（溶血脂）。我们强大的初步数据和出版物 表明：1）我们报道了一个新的概念，大多数溶血脂是新的条件经典损伤- 相关分子模式（DAMP）不结合经典DAMP受体，但结合其 我们的代谢组学数据显示，lysoPI显著地抑制了炎症反应。 高脂饮食（HF）喂养3周的ApoE-/-小鼠的血浆和睾丸中的lysoPI增加; 特异性结合其受体，G蛋白偶联受体55（GPR 55），并激活原代人 主动脉内皮细胞（HAEC）通过上调EC粘附分子ICAM-1; 4）机制上， lysoPI诱导HAEC中线粒体活性氧（mtROS）的产生，并诱导 促炎蛋白精氨酸甲基转移酶1（PRMT 1）活性; 5）我们的RNA-Seq数据显示， 与lysoPI的相对lysoPC诱导的EC显著不同，lysoPI诱导的EC持续时间较长 通过上调DAMP受体炎性体/半胱天冬酶-1和促炎性因子激活 6）我们建立了ApoE-/-/GPR 55-/- DKO小鼠;并且DKO小鼠具有显著的 与用HF喂养12周的ApoE KO小鼠相比动脉粥样硬化减少;最后， 7)我们产生了EC特异性GPR 55 KO小鼠和PRMT 1 KO小鼠。根据我们初步的证据 数据和出版物，中央 假设 需要测试的是早期高脂血症诱导的lysoPI 通过GPR 55-PRMT 1通路刺激和维持主动脉EC活化，促炎单核细胞 (MC)招募，从而促进动脉粥样硬化。我们将用三个目标来检验这个假设。 目的1将确定lysoPI/GPR 55-PRMT 1通路在HAECs中的表达和功能， 高脂血症刺激和ApoE-/-小鼠的动脉粥样硬化（相关研究）。目标2将审查 lysoPI/GPR 55通过诱导PRMT 1诱导持续的主动脉EC活化的机制 上调-mtROS生成，其进一步激活了EHEC（机制研究）。目标3将 确定EC（EC特异性KO）和其他血管内皮细胞中的lysoPI/GPR 55和PRMT 1的抑制是否 细胞（整体KO）将减少ApoE-/-小鼠中的动脉粥样硬化形成（治疗研究）。

Title: LysoPI/GPR55 pathway promotes endothelial activation, vascular inflammation and atherosclerosis Atherosclerosis and its complications are the leading cause of morbidity and mortality in the US. Novel antiinflammatory therapies are urgently needed to inhibit atherosclerosis. The goal of this project is to determine the roles and mechanisms underlying lysophosphatidylinositol (lysoPI) promoted endothelial cell (EC) activation and atherosclerosis. We published numerous papers on EC activation, atherosclerosis and lysophospholipids (lysolipids). Our strong preliminary data and publications show that: 1) we reported a new concept that most lysolipids are new conditional classical damage- associated molecular pattern (DAMPs) that do not bind to classical DAMP receptors but bind to their own receptors to stimulate inflammation; 2) our metabolomics data showed that lysoPI is significantly increased in the aortas and plasma of ApoE-/- mice fed with high fat diet (HF) for three weeks; 3) lysoPI specifically bind to its receptor, G protein coupled receptor 55 (GPR55), and activates primary human aortic endothelial cells (HAECs) by upregulating EC adhesion molecule ICAM-1; 4) Mechanistically, lysoPI induces generation of mitochondrial reactive oxygen species (mtROS) in HAECs and induces proinflammatory proteinarginine methyltransferase 1(PRMT1) activity; 5) our RNA-Seq data showed that significantly different from that induced by lysoPI's relative lysoPC, lysoPI induces sustained EC activation by upregulating DAMP receptors inflammasomes/caspase-1 and proinflammatory secretomes; 6) we established ApoE-/-/GPR55-/- DKO mice; and DKO mice have significantly decreased atherosclerosis in comparison to that in ApoE KO mice fed with HF for 12 weeks; and finally, 7) we generated EC-specific GPR55 KO mice and PRMT1 KO mice. Based on our strong preliminary data and publications, the central hypothesis to be tested is that early hyperlipidemia-induced lysoPI stimulates and sustains aortic EC activation via a GPR55-PRMT1 pathway, proinflammatory monocyte (MC) recruitment, thereby contributing to atherosclerosis. We will test this hypothesis using three aims. Aim 1 will determine expression and function of lysoPI/GPR55-PRMT1 pathway in HAECs activated by hyperlipidemic stimuli and in aortas of ApoE-/- mice (relevant studies). Aim 2 will examine the mechanisms by which lysoPI/GPR55 induces sustained aortic EC activation via inducing PRMT1 upregulation-mtROS generation, which further prolongs EC activation (mechanistic studies). Aim 3 will determine whether inhibition of lysoPI/GPR55 and PRMT1 in EC (EC-specific KO) and other vascular cells (global KO) would decrease atherogenesis in ApoE-/- mice (therapeutic studies).