Diabetes Complications Research Centre, Conway Institute & UCD School of Medicine, University College Dublin, Dublin 4, Ireland.
St. Vincent's University Hospital, Dublin, Ireland.
Atherosclerosis. 2024 Aug;395:117573. doi: 10.1016/j.atherosclerosis.2024.117573. Epub 2024 May 6.
Activation of vascular smooth muscle cell inflammation is recognised as an important early driver of vascular disease. We have previously identified the let-7 miRNA family as important regulators of inflammation in in vitro and in vivo models of atherosclerosis. Here we investigated a dual statin/let-7d-5p miRNA combination therapy approach to target human aortic SMC (HAoSMC) activation and inflammation.
In vitro studies using primary HAoSMCs were performed to investigate the effects of let-7d-5p miRNA overexpression and inhibition. HAoSMCs were treated with combinations of the inflammatory cytokine tumor necrosis factor-α (TNF-α), and atorvastatin or lovastatin. HAoSMC Bulk RNA-seq transcriptomics of HAoSMCs revealed downstream regulatory networks modulated by let-7d-5p miRNA overexpression and statins. Proteome profiler cytokine array, Western blotting and quantitative PCR analyses were performed on HAoSMCs to validate key findings.
Let-7d-5p overexpression significantly attenuated TNF-α-induced upregulation of IL-6, ICAM1, VCAM1, CCL2, CD68, MYOCD gene expression in HAoSMCs (p<0.05). Statins (atorvastatin, lovastatin) significantly attenuated inflammatory gene expression and upregulated Let-7d levels in HAoSMCs (p<0.05). Bulk RNA-seq analysis of a dual Let-7d-5p overexpression/statin therapy in HAoSMCs revealed that let-7d-5p activation and statins converge on key inflammatory pathways (IL-6, IL-1β, TNF-α, IFN-γ). Let-7d-5p overexpression led to reduced expression of the ox-LDL receptor OLR1, and this was associated with lower ox-LDL uptake in HAoSMCs. In silico analysis of smooth muscle cell phenotypic switching shows that overexpression of let-7d-5p in HAoSMCs maintains a contractile phenotype.
Targeting the Let-7 network alongside statins can modulate HAoSMC activation and attenuate key inflammatory pathway signals.
血管平滑肌细胞炎症的激活被认为是血管疾病的一个重要早期驱动因素。我们之前已经确定 let-7 miRNA 家族是动脉粥样硬化的体外和体内模型中炎症的重要调节因子。在这里，我们研究了一种双重他汀类药物/let-7d-5p miRNA 联合治疗方法，以靶向人主动脉平滑肌细胞 (HAoSMC) 的激活和炎症。
使用原代 HAoSMCs 进行体外研究，以研究 let-7d-5p miRNA 过表达和抑制的影响。用炎症细胞因子肿瘤坏死因子-α (TNF-α) 和阿托伐他汀或洛伐他汀处理 HAoSMCs。HAoSMC 总 RNA-seq 转录组学揭示了 let-7d-5p miRNA 过表达和他汀类药物调节的下游调控网络。对 HAoSMCs 进行蛋白质组生物标志物细胞因子阵列、Western 印迹和定量 PCR 分析，以验证关键发现。
let-7d-5p 过表达显著减弱 TNF-α诱导的 HAoSMCs 中 IL-6、ICAM1、VCAM1、CCL2、CD68、MYOCD 基因表达的上调（p<0.05）。他汀类药物（阿托伐他汀、洛伐他汀）显著减弱 HAoSMCs 中的炎症基因表达并上调 Let-7d 水平（p<0.05）。HAoSMCs 中双重 Let-7d-5p 过表达/他汀类药物治疗的总 RNA-seq 分析表明，let-7d-5p 的激活和他汀类药物在关键炎症途径（IL-6、IL-1β、TNF-α、IFN-γ）上汇聚。let-7d-5p 过表达导致 ox-LDL 受体 OLR1 的表达减少，这与 HAoSMCs 中 ox-LDL 的摄取减少有关。平滑肌细胞表型转换的计算机分析表明，HAoSMCs 中 let-7d-5p 的过表达维持收缩表型。
针对 Let-7 网络和他汀类药物可以调节 HAoSMC 的激活并减弱关键炎症途径信号。
