Abstract
Coronary artery diseases (CADs) continue to be the leading global contributors to multi-morbidity and mortality. Given the significant burden of CADs, there is a critical need to identify novel and effective biomarkers for risk assessment. This study sought to evaluate the potential of serum extracellular vesicle-derived small non-coding RNAs (sncRNAs) as predictive biomarkers for CAD risk. Using next-generation sequencing approach, the levels of extracellular vesicles (EVs)-associated sncRNAs were analysed in serum samples from 91 pre-clinical CAD cases and their matched healthy controls, sourced from the prospective EPICOR cohort. We evaluated the predictive ability of sncRNAs alone and in combination with polygenic risk score (PRS) PGS000329. We identified 44 differentially expressed microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs) (FDR < 0.05), which were then narrowed down to ten significant signals (|log2FC|>0.6) for technical validation. RT-qPCR analysis confirmed the trend of expression for two miRNAs (miR-194-5p and miR-451a) and six piRNAs (piR-20266, piR-23533, piR-27282, piR-28212, piR-1043, piR-619). The ROC curve from a Random Forest model showed a higher discrimination ability of piR-619 and piR-23,533 (AUC = 0.72) compared to the use of traditional risk factors alone (AUC = 0.68). To enhance CAD risk assessment, we integrated genetic data by stratifying the cohort into two groups based on the 80th percentile of the PGS000329. We observed an odds ratio (OR) of 2.8 (95% CI: 1.3–6.4, p = 0.01) using PGS000329 alone. When the model was adjusted to include two piRNAs and smoking status, the OR increased to 3.26 (95% CI: 1.2–9.5, p = 0.02). Even though this study is limited by the absence of an independent replication cohort, these findings suggest that the two piRNAs pattern could contribute to predict the risk of CAD and may provide valuable insights into the underlying pathogenesis of the disease, in particular integrating individual CAD-PRS.
Data availability
The genetic and sequencing samples data that support the findings of this study are available from the European Prospective Investigation into Cancer and Nutrition (EPIC) project, but restrictions apply to the availability of these data, which were used under license for the current study, and so are not publicly available. The produced data are available from Prof. Giuseppe Matullo (giuseppe.matullo@unito.it) upon reasonable request and with permission of the EPIC committee. Raw count matrix of the aligned small RNAs across all sequenced samples is provided in Supplementary material.
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Acknowledgements
The authors acknowledge Giovanni Camussi (Dep of Medical Sciences, University of Turin) for his contribution in characterizing extracellular vesicles.
Funding
EPIC, EPICOR and EPICOR2 projects were supported by the Compagnia di San Paolo for the EPIC, EPICOR and EPICOR2 projects (SP, SS, RT, PV, LI, CS, GM) and from the Ministero dell’Istruzione, dell’Università e della Ricerca—c (n° D15D18000410001, to G.M.) to the Department of Medical Sciences, University of Torino. This work was also supported by the “Genoma mEdiciNa pERsonalizzatA – GENERA”, funded from the Ministero dell’Istruzione, dell’Università e della Ricerca 2021 (n° D73C22000960001) and by CARdiomyopathy in type 2 DIAbetes mellitus (Cardiateam) funded from the Innovative Medicines Initiative 2 Joint Undertaking (JU) under grant agreement No 821508.
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E.C. G.M. A.A. C.S. conceptualised the study. E.C. G.M. A.A. and C.C initiated and designed the experiments. G.M. P.V. and C.S. help in the selection of the samples. E.C. M.R. and C.D. wrote the main manuscript text. G.B. S.A. C.D. C.D.P. E.C. performed the bioinformatic and statistical analyses. A.P. C.G. and G.A. revised the manuscript.All authors gave final approval of the version to be published. All authors made a significant contribution to the work reported.
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The European Prospective Investigation into Cancer and Nutrition (EPIC) study protocol was approved by the ethics committees centralized at the International Agency for Research on Cancer (Lyon, France) (reference number: IEC 24 − 08). The EPICOR Study, a case–cohort study nested within the EPIC–Italy prospective cohort, was approved by the HuGeF Ethics Committee in Turin on 15 December 2010.
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Casalone, E., Rosselli, M., Birolo, G. et al. Integration of short non coding RNA and genetic factors for coronary artery disease risk prediction in a prospective study. Sci Rep (2026). https://doi.org/10.1038/s41598-026-38355-4
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DOI: https://doi.org/10.1038/s41598-026-38355-4
