Inhibition of vascular smooth muscle cell (VSMC) proliferation by intracellular cAMP prevents excessive neointima formation and hence angioplasty restenosis and vein-graft failure. These protective effects are mediated via actin-cytoskeleton remodelling and subsequent regulation of gene expression by mechanisms that are incompletely understood. Here we investigated the role of components of the growth-regulatory Hippo pathway, specifically the transcription factor TEAD and its co-factors YAP and TAZ in VSMC. Elevation of cAMP using forskolin, dibutyryl-cAMP or the physiological agonists, Cicaprost or adenosine, significantly increased phosphorylation and nuclear export YAP and TAZ and inhibited TEAD-luciferase report gene activity. Similar effects were obtained by inhibiting RhoA activity with C3-transferase, its downstream kinase, ROCK, with Y27632, or actin-polymerisation with Latrunculin-B. Conversely, expression of constitutively-active RhoA reversed the inhibitory effects of forskolin on TEAD-luciferase. Forskolin significantly inhibited the mRNA expression of the pro-mitogenic genes, CCN1, CTGF, c-MYC and TGFB2 and this was reversed by expression of constitutively-active YAP or TAZ phospho-mutants. Inhibition of YAP and TAZ function with RNAi or Verteporfin significantly reduced VSMC proliferation. Furthermore, the anti-mitogenic effects of forskolin were reversed by overexpression of constitutively-active YAP or TAZ. Taken together, these data demonstrate that cAMP-induced actin-cytoskeleton remodelling inhibits YAP/TAZ-TEAD dependent expression of pro-mitogenic genes in VSMC. This mechanism contributes novel insight into the anti-mitogenic effects of cAMP in VSMC and suggests a new target for intervention.

Fetal growth restriction (FGR) is a condition where the fetus does not reach its genetically predetermined size, affecting 1 in 10 pregnancies and contributing to up to 50% of all stillbirths before 34 weeks of gestation. Current diagnostic methods primarily involve ultrasound and Doppler assessments, yet there is growing interest in identifying biomarkers for early diagnosis and improved management. This systematic review examined the role of microRNAs (miRNAs) in the pathogenesis of FGR, focusing on their potential as non-invasive biomarkers. MicroRNAs are small, non-coding RNAs that regulate gene expression. This review systematically assessed studies investigating the differential expression of miRNAs in maternal blood, serum, and plasma samples from FGR-affected pregnancies. A total of nine studies met the inclusion criteria, which showed the differential expression of a total of 48 miRNAs. miR-16-5p was consistently upregulated in multiple studies and trimesters. miR-590-3p and miR-206 were consistently upregulated in multiple trimesters. The common gene targets of these miRNAs are VEGF, PIGF, and MMP9. The downregulation of these genes contributes to impaired angiogenesis, trophoblast invasion, placental function, and fetal growth.

Physical activity (PA) and body composition are important lifestyle factors that influence public health. Research suggests that DNA regions (CpG site locations) are differentially methylated in a physically active population. This meta-analysis aimed to identify CpG sites associated with various levels of PA and associated metabolic pathways. The meta-analysis followed PRISMA guidelines using PubMed, SportDISCUS, Embase, Scopus, Cochrane and Web of Science. Epigenomic analyses performed on DNA of participants with no underlying health conditions were included. Articles were screened using Rayyan AI and extracted CpG sites, and their location were confirmed using the EWAS catalogue. Six studies comprising 770 subjects were included in this meta-analysis. The meta-analysis was performed on clinical metrics extracted from the six studies and showed that BMI, blood pressure, insulin and glucose testing are significantly improved upon PA intervention. Amongst the included studies, a total of 257 CpG sites were differentially methylated in physically active participants, with 134 CpGs located in 92 genes associated with obesity-related pathways. The identified differentially methylated genes either belonged to the lipid metabolism or insulin signalling pathway. The genes which were differentially regulated in multiple tissue types and studies are JAZF1 (insulin signalling, and lipid and carbohydrate metabolism pathways) and NAV1 (mTOR signalling pathway). In conclusion, the current epigenomic meta-analysis showed that PA levels induce differential DNA methylation signatures on genes that affect metabolism. To understand the positive molecular effects of PA, further research on the above candidate genes needs to be conducted amongst various levels of a physically active population.

Aim: The aim of this study was to correlate single-nucleotide polymorphisms (SNPs) in the FTO and MC4R genes with body composition (BC) in populations with various levels of physical activity, and to investigate associations of SREBF1 methylation with the level of physical activity (PA) and BC. Methods: Fifty-six participants aged 18–65 years old with no underlying medical conditions were included in the study and were classified into sedentary/light PA (SLPA), moderate PA (MPA) and vigorous PA (VPA) groups using the International PA questionnaire (IPAQ). Anthropometric measures such as age, gender, body mass index (BMI) and body fat percentage (BFP) were recorded at the time of recruitment. Venous blood samples were collected during participant recruitment and DNA was extracted. Genotyping assays were performed for SNPs in FTO (rs9939609) and MC4R (rs17782313) using Taqman^® RT qPCR and TaqMan Genotyper software 1.7.1. Methylation analysis assay for CpG sites in the SREBF1 gene was performed on 56 samples using PyroMark^® Q48 Autoprep (Qiagen, Venlo, The Netherlands). The results were statistically analysed to identify any associations between FTO/MC4R genotypes and the level of PA, and between SREBF1 methylation status and the level of PA. This is the first study to investigate links between PA and quantitative methylation of SREBF1. Results: According to IPAQ guidance, the 56 participants were classified into SLPA n = 14, MPA n = 11 and VPA n = 31. The correlation analysis revealed that the FTO rs9939609 ‘A’ risk allele had a significant negative association with BFP in the VPA group (p = 0.0387); the MC4R rs17782313 ‘C’ risk allele had a significant positive association with BMI in the VPA group (p = 0.0256). In the SREBF1 pyrosequencing analysis, higher levels of methylation were observed in the VPA group (p = 0.07). Conclusions: We concluded that SNPs associated with obesity identified in FTO rs9939609 and MC4R rs17782313 could help to predict the molecular effects of PA. A high frequency of FTO risk variants in the cohort was observed and the VPA group could help maintain a healthy BFP.

Background and AimsLiraglutide a multifaceted drug prescribed in weight management and control of type 2 diabetes. Clinical trials demonstrate that liraglutide significantly reduces cardiovascular-related mortality. The purpose of this study is to explore the potential molecular mechanism of liraglutide in alleviating the effects of hyperglycaemia in VSMCs.Methods and ResultsTotal RNA was extracted from vascular smooth muscle cells (VSMCs) subjected to four experimental conditions: untreated control, high glucose (HG; 25 mM), liraglutide (LIRA; 100 nM), and combined liraglutide plus high glucose (LIRA+HG). Transcriptome profiling was performed using high-throughput RNA sequencing to assess differential gene expression across these treatment groups. Transcriptomics data followed by gene enrichment analysis identified GSDMC, MT1G and BRSK2 as differentially expressed mRNAs in the four treatment groups. Further analysis highlighted significantly down- and upregulated DE-mRNAs involved in MAPK, PI3K/Akt, mTOR, and VEGF pathways, which are associated with cardiovascular protective effects of LIRA in VSMCs. Western blot and RT-qPCR results revealed that LIRA downregulated AKT1, MAP2K1, PDK1, and PI3K, indicating suppression of PI3K/Akt-mediated proliferative signalling. Caspase activity, cell proliferation, and migration assays performed on all four groups demonstrate that liraglutide significantly attenuates HG-induced proliferation and migration and restores apoptosis levels.ConclusionGene expression of known proliferative pathways, including MAPK, PI3K/Akt, mTOR, and VEGF is reduced in VSMC treated with LIRA and LIRA+HG compared to control and HG groups. Transcriptomics data highlights novel DE-mRNAs GSDMC, MT1G and BRSK2 which represent potential cardioprotective targets, warranting further experimental validation and clinical investigation to elucidate their functional roles in vascular pathology.

Background/Objectives: Metabolic/bariatric surgery is currently the most successful treatment for patients with obesity; however, a fifth of patients undergoing surgery may not lose enough weight to be considered successful. Recent studies have shown that bariatric/metabolic surgery alters the epigenome and may explain postoperative improvements in metabolic health. The primary objective is to consolidate published differentially methylated CpG sites in pre- and post-metabolic/bariatric surgery female patients and associate them with the respective genes and pathways. Methods: This systematic review adhered to the PRISMA-P guidelines and was registered with the PROSPERO (CRD42023421852). Following an initial screening of 541 studies using COVIDENCE, six studies were selected, comprising three epigenome-wide association studies (EWAS) and three candidate gene methylation studies. The published studies collected DNA samples from female patients with obesity before and after surgery (3 months, 6 months, 9–31 months, and 2 years). KEGG pathway analysis was performed on genes where the extracted CpG sites were located. Results: The meta-analysis showed that 11,456 CpG sites are differentially methylated after a successful weight loss surgery, with 109 sites mapped to genes involved in key metabolic pathways, including FoxO, mTOR, insulin, cAMP, adipocytokine, Toll-like receptor, and PI3K-Akt. Conclusion: The highlighted differentially methylated CpG sites can be further used to predict the molecular signature associated with successful metabolic/bariatric surgery.