The role of macrophage VCAM-1 in atherosclerosis and inflammation

VCAM-1 is known to be expressed by endothelial cells. We showed that this is also expressed by atherosclerotic plaque macrophages. However, the merit of this adhesion molecule is now known in atherosclerosis. In this RNA sequencing experiment, we analyzed VCAM-1+ vs. - atherosclerotic plaque macrophages. Additionally, we sorted macrophages from atherosclerotic plaques of mice after mitochondrial biogenesis gene Cmpk2 silencing in macrophages in vivo. Overall design: We sorted VCAM-1+ and - macrophages from atherosclerotic plaques of Apoe KO mice fed with an atherogenic diet for four months. Moreover, we collected total atherosclerotic plaque macrophages from ApoeKO mice fed with an atherogenic diet for four months and treated with control and Cmpk2 siRNA formulated in lipidoid nanoparticles for a month. For the spatial transcriptomics experiment, we performed bone marrow transplantation in Ldlr KO mice with bone marrow from WT or myeloid Vcam1-deficient mice. The mice were placed on high fat diet a week later. The aortic roots were harvested after four months, spatial RNA seq using the Slide Seq technology was employed.

已知血管细胞黏附分子1（VCAM-1）可由内皮细胞表达。本研究证实，动脉粥样硬化斑块巨噬细胞同样表达该分子。尽管目前学界已明确该黏附分子在动脉粥样硬化中的生物学功能，本RNA测序实验仍对VCAM-1阳性与VCAM-1阴性的动脉粥样硬化斑块巨噬细胞进行了对比分析。此外，我们在体内对巨噬细胞的线粒体生物发生相关基因Cmpk2实施沉默后，从小鼠动脉粥样硬化斑块中分离分选了巨噬细胞。
整体实验设计如下：我们从喂食致动脉粥样硬化饮食4个月的载脂蛋白E敲除（Apoe KO）小鼠的动脉粥样硬化斑块中，分选得到VCAM-1阳性与阴性巨噬细胞。同时，我们从同样喂食致动脉粥样硬化饮食4个月的Apoe KO小鼠中收集总动脉粥样硬化斑块巨噬细胞，并分别用对照小干扰RNA（siRNA）以及包载于脂质纳米颗粒中的Cmpk2 siRNA对小鼠进行为期1个月的处理。
针对空间转录组学实验，我们对低密度脂蛋白受体敲除（Ldlr KO）小鼠开展骨髓移植，供体骨髓分别取自野生型（WT）小鼠或髓系特异性Vcam1缺陷小鼠。骨髓移植一周后，小鼠被喂食高脂饮食。造模4个月后收取小鼠主动脉根，采用Slide Seq技术（Slide Seq technology）进行空间RNA测序。