Dr. B R Ambedkar Center for Biomedical Research, University of Delhi, New Delhi, 110007, India.
Department of Cardiology, All India Institute of Medical Sciences, New Delhi, 110029, India.
Mol Diagn Ther. 2022 Nov;26(6):655-664. doi: 10.1007/s40291-022-00615-0. Epub 2022 Oct 5.
Atherosclerosis, a progressive manifestation of coronary artery disease, has been observed to be regulated by microRNAs (miRNAs) targeting various protein-coding genes involved in several pathophysiological events of coronary artery disease.
In our previous report, we identified differential expression profiles of candidate miRNAs, miR-133b and miR-21, in patients with coronary artery disease as compared with controls, suggesting their possible implication in the pathophysiology of coronary artery disease. To better understand the functional role of these miRNAs, we sought to predict and validate their target genes while assessing the expression pattern of these genes in patients with coronary artery disease, as well as in macrophages.
Potential target genes of miR-133b and miR-21 were predicted bioinformatically followed by validation through the identification of their expression at the protein level in patients with coronary artery disease (n-30), as well as in macrophages treated with respective miRNA inhibitors (antagomiRs), through immunoblotting.
SGPP1, a gene associated with the sphingolipid pathway, was predicted to be a potential target gene of miR-133b while ATG5 and LRP6 were target genes of miR-21 while being associated with autophagy and Wnt signalling pathways, respectively. SGPP1 was observed to be upregulated significantly (p = 0.019) by 2.07-fold, whereas ATG5 and LRP6 were found to be downregulated (p = 0.026 and 0.007, respectively) by 3.28-fold and 8.46-fold, respectively, in patients with coronary artery disease as compared with controls. Expression patterns of all the genes were also found to be modulated when cells were treated with respective miRNA inhibitors.
Results from the present study suggest that SGPP1, ATG5 and LRP6, target genes of miR-133b and miR-21, may serve as potential therapeutic hotspots in the management of coronary artery disease, which undoubtedly merit further experimental confirmation.
动脉粥样硬化是冠状动脉疾病的一种进行性表现，现已观察到其受针对参与冠状动脉疾病几种病理生理事件的各种蛋白编码基因的 microRNAs（miRNAs）调控。
在我们之前的报告中，我们发现与对照组相比，冠状动脉疾病患者候选 miRNA（miR-133b 和 miR-21）的差异表达谱，提示它们可能在冠状动脉疾病的病理生理学中发挥作用。为了更好地了解这些 miRNA 的功能作用，我们试图预测和验证它们的靶基因，同时评估这些基因在冠状动脉疾病患者以及巨噬细胞中的表达模式。
通过生物信息学预测 miR-133b 和 miR-21 的潜在靶基因，然后通过鉴定它们在冠状动脉疾病患者（n=30）中的蛋白水平表达，以及用相应的 miRNA 抑制剂（antagomiRs）处理的巨噬细胞中的表达来验证。
与鞘脂代谢途径相关的 SGPP1 基因被预测为 miR-133b 的潜在靶基因，而 ATG5 和 LRP6 则分别是 miR-21 的靶基因，与自噬和 Wnt 信号通路相关。与对照组相比，冠状动脉疾病患者的 SGPP1 显著上调（p=0.019），上调 2.07 倍，而 ATG5 和 LRP6 下调 3.28 倍和 8.46 倍（p=0.026 和 0.007）。用相应的 miRNA 抑制剂处理细胞时，所有基因的表达模式也被发现发生了调节。
本研究结果表明，miR-133b 和 miR-21 的靶基因 SGPP1、ATG5 和 LRP6 可能成为冠状动脉疾病治疗的潜在热点，这无疑值得进一步的实验验证。
