Sukhorukov Vasily Nikolaevich, Khotina Victoria Alexandrovna, Borodko Daria Dmitryevna, Ekta Mariam Bagheri, Oishi Yumiko, Omelchenko Andrey Vladimirovich, Kolmychkova Kira Ivanovna, Nikiforov Nikita G, Sobenin Igor Alexandrovich, Orekhov Alexander Nikolaevich
Laboratory of Angiopathology, Institute of General Pathology and Pathophysiology, 125315, Moscow, Russia.
Laboratory of Cellular and Molecular Pathology of Cardiovascular System, Petrovsky National Research Centre of Surgery, 119991, Moscow, Russia.
Curr Med Chem. 2025;32(9):1755-1769. doi: 10.2174/0109298673286400240206095814.
The relationship between the cellular pro-inflammatory response and intracellular lipid accumulation in atherosclerosis is not sufficiently studied. Transcriptomic analysis is one way to establish such a relationship. Previously, we identified 10 potential key genes (IL-15, CXCL8, PERK, IL-7, IL-7R, DUSP1, TIGIT, F2RL1, TSPYL2, and ANXA1) involved in cholesterol accumulation in macrophages. It should be noted that all these genes do not directly participate in cholesterol metabolism, but encode molecules related to inflammation.
In this study, we conducted a knock-down of the 10 identified key genes using siRNA to determine their possible role in cholesterol accumulation in macrophages. To assess cholesterol accumulation, human monocyte-derived macrophages (MDM) were incubated with atherogenic LDL from patients with atherosclerosis. Cholesterol content was assessed by the enzymatic method. Differentially expressed genes were identified with DESeq2 analysis. Master genes were determined by the functional analysis.
We found that only 5 out of 10 genes (IL-15, PERK, IL-7, IL-7R, ANXA1) can affect intracellular lipid accumulation. Knock-down of the IL-15, PERK, and ANXA1 genes prevented lipid accumulation, while knock-down of the IL-7 and IL-7R genes led to increased intracellular lipid accumulation during incubation of MDM with atherogenic LDL. Seventeen overexpressed genes and 189 underexpressed genes were obtained in the DGE analysis, which allowed us to discover 20 upregulated and 86 downregulated metabolic pathways, a number of which are associated with chronic inflammation and insulin signaling. We also elucidated 13 master regulators of cholesterol accumulation that are immune response-associated genes.
Thus, it was discovered that 5 inflammation-related master regulators may be involved in lipid accumulation in macrophages. Therefore, the pro-inflammatory response of macrophages may trigger foam cell formation rather than the other way around, where intracellular lipid accumulation causes an inflammatory response, as previously assumed.
细胞促炎反应与动脉粥样硬化中细胞内脂质积累之间的关系尚未得到充分研究。转录组分析是建立这种关系的一种方法。此前，我们鉴定出10个参与巨噬细胞胆固醇积累的潜在关键基因（IL-15、CXCL8、PERK、IL-7、IL-7R、DUSP1、TIGIT、F2RL1、TSPYL2和ANXA1）。需要注意的是，所有这些基因并不直接参与胆固醇代谢，而是编码与炎症相关的分子。
在本研究中，我们使用小干扰RNA（siRNA）敲低这10个已鉴定的关键基因，以确定它们在巨噬细胞胆固醇积累中的可能作用。为了评估胆固醇积累情况，将人单核细胞衍生的巨噬细胞（MDM）与动脉粥样硬化患者的致动脉粥样硬化低密度脂蛋白一起孵育。通过酶法评估胆固醇含量。使用DESeq2分析鉴定差异表达基因。通过功能分析确定主基因。
我们发现10个基因中只有5个（IL-15、PERK、IL-7、IL-7R、ANXA1）会影响细胞内脂质积累。敲低IL-15、PERK和ANXA1基因可防止脂质积累，而敲低IL-7和IL-7R基因则导致MDM与致动脉粥样硬化低密度脂蛋白孵育期间细胞内脂质积累增加。在差异基因表达分析中获得了17个过表达基因和189个低表达基因，这使我们发现了20条上调和86条下调的代谢途径，其中许多与慢性炎症和胰岛素信号传导相关。我们还阐明了13个胆固醇积累的主调节因子，它们是免疫反应相关基因。
因此，发现5个与炎症相关的主调节因子可能参与巨噬细胞中的脂质积累。因此，巨噬细胞的促炎反应可能触发泡沫细胞形成，而不是像之前所认为的那样，细胞内脂质积累引起炎症反应。
