| Summary: | Chronic limb-threatening ischemia (CLTI) results in devastating complications such as lower-limb amputations. In this study, a genome-wide plasma microRNAs (miRNA) sequencing was performed to identify miRNA(s) associated with CLTI. Blood samples were collected from early stage CLTI patients (ABI < 0.9) and non-PAD controls (ABI ≥ 0.9) for 3 experiments: discovery phase (<i>n</i> = 23), confirmatory phase (<i>n</i> = 52) and validation phase (<i>n</i> = 20). In the discovery phase, next generation sequencing (NGS) was used to identify miRNA circulating in the plasma CLTI (<i>n</i> = 13) patients, compared to non-PAD controls (<i>n</i> = 10). Two down-regulated miRNAs (miRNA-6843-3p and miRNA-6766-5p) and three upregulated miRNAs (miRNA-1827, miRNA-320 and miRNA-98-3p) were identified (≥2-fold change). In the confirmatory phase, these 5 deregulated miRNAs were further investigated in non-PAD (<i>n</i> = 21) and CTLI (<i>n</i> = 31) patients using qRT-PCR. Only miRNA-1827 was found to be significantly upregulated (≥3-fold, <i>p</i>-value < 0. 001) in the CLTI group. Lastly, to minimize the influence of confounding factors, miRNA-1827 plasma levels were validated in a third cohort of CLTI patients (<i>n</i> = 10) matched to non-PAD controls (<i>n</i> = 10). Our analysis demonstrated that miRNA-1827 expression was increased in the CLTI cohort (≥2-folds, <i>p</i>-value < 0.001). In summary, circulating miRNA-1827 is significantly elevated in patients with CLTI.
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