ASSOCIATION OF MICRO RIBONUCLEIC ACID (miRNA) 654-5P EXPRESSION WITH COLLATERAL DEGREE IN CHRONIC LIMB THREATENING ISCHEMIA

Abstract

Background: Chronic Limb-Threatening Ischemia (CLTI) represents most severe form of chronic peripheral arterial disease, which is associated with poor clinical outcomes if left untreated. Despite advances in interventional and pharmacological therapies, peripheral arterial disease remains a major public health issue. Several studies have reported that miRNA play a role in arteriogenesis through multiple signaling pathways. One of the novel miRNAs identified through profiling studies is miRNA 654-5p. However, literature specifically analysing miRNA 654-5p as a biomarker and its association with CLTI remains very limited, and no previous studies have linked its expression levels with the degree of collateral formation in CLTI.

Objectives: The primary objective of this study was to analyze the difference in miRNA 654-5p expression between patients with and without CLTI, as well as according to the degree of collateral formation among patients treated at Dr. Sardjito General Hospital. The secondary objective was to determine the differences in collateral degree based on age, sex, hypertension, diabetes mellitus, smoking status, dyslipidemia, and chronic kidney disease.

Methods: This was an analytical observational study with a cross-sectional design using secondary data from a previous study by Ismail et al. (2025), conducted from February to October 2023. The study included adult patients diagnosed with CLTI according to the 2019 ESVS Guidelines on the Management of Chronic Limb-Threatening Ischemia. These subjects had undergone arteriography and blood sample was taken for miR-654-5p expression analysis.

Results: A total of 33 subjects were included, consisting of two groups: CLTI (n = 15) and non-CLTI (n = 18). miRNA 654-5p expression was found to be upregulated with a fold change value of 3.45. The ROC curve analysis yielded an AUC of 57%, with an optimal cut-off point based on the Youden index was 28.56 with 80% sensitivity and 50% specificity (p = 0.492, 95% CI 0.370–0.771). Based on this cut-off, there was no statistically significant association between miRNA 654-5p expression and the presence of CLTI (OR 4; 95% CI 0.835–19.162; p = 0.74). However, a significant difference was found between miRNA 654-5p expression and collateral grade (p = 0.001). On bivariate analysis, the association between collateral grade and confounders were not statistically significant.

Conclusion: miRNA 654-5p expressions were higher in subjects with CLTI, although with weak statistical power. Higher miRNA 654-5p expression were higher in subjects with a worse collateral grade.