Hippo signalling-linked circulating microRNA networks associate with cardiovascular disease in Type 2 Diabetes

Abstract Context Type 2 Diabetes (T2D) markedly increases the risk of cardiovascular disease (CVD). Circulating microRNAs (miRNAs) are promising biomarkers for patient stratification and may provide mechanistic insight into dysregulated pathways in T2D-associated CVD. Objective To identify circulating miRNA signatures in T2D patients with CVD (T2D-CVD) versus T2D patients without CVD using a novel network-based medicine approach. Methods Patients (n = 110) were stratified into T2D and T2D-CVD groups based on the absence or presence of macrovascular complications. Circulating miRNAs, pro-inflammatory cytokines and growth factors were profiled in a discovery cohort (n = 80) and confirmed in a validation cohort (n = 30). Differential expression, network-based approaches and pathway enrichment analyses identified miRNA signatures, co-expression communities, hub miRNAs, and associated signalling pathways. Results T2D-CVD patients showed a set of elevated inflammatory cytokines and circulating miRNAs. Particularly, nineteen miRNAs were differentially expressed between T2D-CVD and T2D patients. Pathway enrichment of these miRNAs highlighted Hippo signalling together with TGF-β, mTOR, Wnt, and PI3K–Akt pathways. Network analysis identified a co-expression community of 34 miRNAs, including hub miRNAs let-7a-5p, miR-132-3p, and miR-1306-3p, all linked to Hippo signalling. Key miRNA interactions were validated, and both differentially expressed and hub miRNAs correlated with inflammatory mediators such as PAR-1, CXCL1 and NEMO, which are linked to Hippo pathway regulation. Conclusions This study identified a network of circulating miRNAs associated with inflammatory cytokines and Hippo signalling pathway in T2D-CVD. These findings reveal novel molecular aspects of the systemic inflammation characterizing T2D-CVD and support the potential of Hippo-related circulating miRNAs as mechanistic biomarkers for cardiovascular risk in T2D.