Exploration of dual LET-7 MIMIC/statin therapy targeting vascular smooth muscle cell dysfunction in atherosclerosis

Background and Aims : Micro-RNA (miRNA) act as negative post-transcriptional regulators of gene expression and are involved in every stage of atherosclerotic plaque development. Statins remain the most common therapeutic agent for lipid lowering, and there is great interest in understanding the pleiotropic effects of statins as well as investigating drug combination therapies alongside statins. Here we investigated a dual statin/miRNA combination therapy approach to target human aortic SMC (HAoSMC) activation.Methods: miRNA-mRNA network enrichment analysis was performed in human atherosclerotic plaque transcriptomics datasets using MiRNET 2.0. Primary HAoSMCs (Lonza Bioscience) were transfected with combinations of Let-7d mimic, miR-27a mimic and miR-155 antagomiR (20nmol/L;24h) and treated with pro-atherogenic stimuli (TNF-α, 10ng/ml;24h), and Atorvastatin (1μM;24h) or Lovastatin (1μM;24h). RNA-seq transcriptomics was performed by the Beijing Genomics Institute (DNBseq). All cell experiments were performed n=3-5 times.Results: Analysis of miRNA-gene network interactions in human atherosclerosis transcriptomics datasets identified lead miRNA networks (miRs-27a, -33, -155, - 16 and Let-7d). Let-7d mimic significantly attenuated TNF-α-induced increase of IL-6, ICAM-1, VCAM-1, MCP1, CD68, MYOCD gene expression (p<0.05) in HAoSMCs. Statins (Atorvastatin, Lovastatin) significantly attenuated inflammatory gene expression and increased the expression of Let-7d in HAoSMCs (p<0.05). We next investigated dual Let-7d mimic/statin therapy in HAoSMCs treated with TNF-α. Our preliminary findings indicate that miRNA modulating strategies can enhance the effects of statins in HAoSMCs. RNA-seq studies elucidated transcriptome-wide responses to these therapies, identifying multiple inflammatory pathways modulate by these therapies.Conclusions: Targeting the Let-7 network alongside statins can modulate HAoSMC activation and attenuate key inflammatory pathway signals. Background and Aims : Micro-RNA (miRNA) act as negative post-transcriptional regulators of gene expression and are involved in every stage of atherosclerotic plaque development. Statins remain the most common therapeutic agent for lipid lowering, and there is great interest in understanding the pleiotropic effects of statins as well as investigating drug combination therapies alongside statins. Here we investigated a dual statin/miRNA combination therapy approach to target human aortic SMC (HAoSMC) activation. Methods: miRNA-mRNA network enrichment analysis was performed in human atherosclerotic plaque transcriptomics datasets using MiRNET 2.0. Primary HAoSMCs (Lonza Bioscience) were transfected with combinations of Let-7d mimic, miR-27a mimic and miR-155 antagomiR (20nmol/L;24h) and treated with pro-atherogenic stimuli (TNF-α, 10ng/ml;24h), and Atorvastatin (1μM;24h) or Lovastatin (1μM;24h). RNA-seq transcriptomics was performed by the Beijing Genomics Institute (DNBseq). All cell experiments were performed n=3-5 times. Results: Analysis of miRNA-gene network interactions in human atherosclerosis transcriptomics datasets identified lead miRNA networks (miRs-27a, -33, -155, - 16 and Let-7d). Let-7d mimic significantly attenuated TNF-α-induced increase of IL-6, ICAM-1, VCAM-1, MCP1, CD68, MYOCD gene expression (p<0.05) in HAoSMCs. Statins (Atorvastatin, Lovastatin) significantly attenuated inflammatory gene expression and increased the expression of Let-7d in HAoSMCs (p<0.05). We next investigated dual Let-7d mimic/statin therapy in HAoSMCs treated with TNF-α. Our preliminary findings indicate that miRNA modulating strategies can enhance the effects of statins in HAoSMCs. RNA-seq studies elucidated transcriptome-wide responses to these therapies, identifying multiple inflammatory pathways modulate by these therapies. Conclusions: Targeting the Let-7 network alongside statins can modulate HAoSMC activation and attenuate key inflammatory pathway signals.