miR-33 Silencing Reprograms the Immune Cell Landscape in Atherosclerotic Plaques.
miR-33 Silencing Reprograms the Immune Cell Landscape in Atherosclerotic Plaques.
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miR-33沉默重新编程动脉粥样硬化斑块中的免疫细胞景观。
DOI：
10.1161/circresaha.120.317914
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发表时间：
2021-04-16
影响因子：
20.1
通讯作者：
Moore KJ
中科院分区：
文献类型：
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作者：
Afonso MS;Sharma M;Schlegel M;van Solingen C;Koelwyn GJ;Shanley LC;Beckett L;Peled D;Rahman K;Giannarelli C;Li H;Brown EJ;Khodadadi-Jamayran A;Fisher EA;Moore KJ
MicroRNA-33转录后抑制与脂肪代谢和能量平衡有关的基因。靶向抑制miR-33可增加血浆高密度脂蛋白胆固醇，并促进动脉粥样硬化的消退，部分是通过加强胆固醇的反向转运和抑制斑块炎症。然而，miR-33如何重塑斑块的免疫微环境仍知之甚少。明确miR-33抑制如何改变动脉粥样硬化斑块中免疫细胞的动态平衡和转录格局。我们通过对主动脉CD45+细胞进行单细胞RNA测序，结合免疫组织学、形态计量学和流式细胞术分析，确定miR-33抑制后斑块免疫细胞组成、基因表达和功能的变化。我们报道，抗miR-33治疗晚期动脉粥样硬化的LDLR−/−小鼠减少了斑块负荷，并通过改变支持和抗动脉粥样硬化的巨噬细胞和T细胞亚群的平衡改变了斑块免疫细胞格局。通过量化决定斑块巨噬细胞负荷的动力学过程，我们发现抗miR-33降低了循环单核细胞和脾髓系祖细胞的水平，减少了巨噬细胞的增殖和滞留，并通过凋亡和吞噬作用促进了巨噬细胞的磨损。主动脉弓斑块的scRNA测序显示，抗miR-33可降低MHCIIHI“炎性”和Trem2HI“代谢”巨噬细胞的频率，但不能减少组织驻留巨噬细胞。此外，在不同的巨噬细胞亚群中，抗miR-33抗体导致不同的miR-33靶基因的表达下调：在常驻和Trem2hi巨噬细胞中，抗miR-33抗体缓解了涉及脂质代谢的miR-33靶基因(如ABCA1、Ncoa1、NCOA2、CROT)的抑制，而在MHCIIhi巨噬细胞中，抗miR-33抗体上调了参与染色质重塑和转录调节的靶基因。抗miR-33还减少了主动脉CD8+T细胞和CD4+Th1细胞的积聚，增加了斑块中FoxP3+调节性T细胞的水平，这与免疫抑制斑块炎症的作用一致。我们的结果为了解斑块中执行抗MIR-33‘S动脉粥样硬化保护作用的免疫机制和细胞因子提供了深入的了解。
MicroRNA-33 post-transcriptionally represses genes involved in lipid metabolism and energy homeostasis. Targeted inhibition of miR-33 increases plasma HDL cholesterol and promotes atherosclerosis regression, in part, by enhancing reverse cholesterol transport and dampening plaque inflammation. However, how miR-33 reshapes the immune microenvironment of plaques remains poorly understood. To define how miR-33 inhibition alters the dynamic balance and transcriptional landscape of immune cells in atherosclerotic plaques. We used single cell RNA-sequencing of aortic CD45+ cells, combined with immunohistologic, morphometric and flow cytometric analyses to define the changes in plaque immune cell composition, gene expression and function following miR-33 inhibition. We report that anti-miR-33 treatment of Ldlr−/− mice with advanced atherosclerosis reduced plaque burden and altered the plaque immune cell landscape by shifting the balance of pro- and anti-atherosclerotic macrophage and T cell subsets. By quantifying the kinetic processes that determine plaque macrophage burden, we found that anti-miR-33 reduced levels of circulating monocytes and splenic myeloid progenitors, decreased macrophage proliferation and retention, and promoted macrophage attrition by apoptosis and efferocytotic clearance. scRNA-sequencing of aortic arch plaques showed that anti-miR-33 reduced the frequency of MHCIIhi “inflammatory” and Trem2hi “metabolic” macrophages, but not tissue resident macrophages. Furthermore, anti-miR-33 led to derepression of distinct miR-33 target genes in the different macrophage subsets: in resident and Trem2hi macrophages, anti-miR-33 relieved repression of miR-33 target genes involved in lipid metabolism (e.g., Abca1, Ncoa1, Ncoa2, Crot), whereas in MHCIIhi macrophages, anti-miR-33 upregulated target genes involved in chromatin remodeling and transcriptional regulation. Anti-miR-33 also reduced the accumulation of aortic CD8+ T cells and CD4+ Th1 cells, and increased levels of FoxP3+ regulatory T cells in plaques, consistent with an immune-dampening effect on plaque inflammation. Our results provide insight into the immune mechanisms and cellular players that execute anti-miR-33’s atheroprotective actions in the plaque.