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Updated: Feb 26, 2026
Digital PCR for Quantifying Circulating MicroRNAs in Acute Myocardial Infarction and Cardiovascular Disease
Published on: July 3, 2018
RNA-Seq Identifies Circulating miR-125a-5p, miR-125b-5p, and miR-143-3p as Potential Biomarkers for Acute Ischemic
Steffen Tiedt1, Matthias Prestel1, Rainer Malik1
1From the Institute for Stroke and Dementia Research, Klinikum der Universität München (S.T., M.P., R.M., N.S., M.D., V.K., I.S., J.B., A.L., N.P., M.D.), Graduate School of Systemic Neurosciences (S.T.), Institute of Laboratory Medicine, Klinikum der Universität München (B.H.N., D.T., L.M.H.), Department of Neurology, Klinikum der Universität München (M.K.), and Department of Neuroradiology, Klinikum der Universität München (F.D.), Ludwig-Maximilians-Universität LMU, Germany; Munich Cluster for Systems Neurology (SyNergy), Germany (S.T., A.L., N.P., M.D.); and Interdisciplinary Center for Clinical Research (IZKF), University of Leipzig, Germany (K.K.).
Rationale:
Currently, there are no blood-based biomarkers with clinical utility for acute ischemic stroke (IS). MicroRNAs show promise as disease markers because of their cell type-specific expression patterns and stability in peripheral blood.
Objective:
To identify circulating microRNAs associated with acute IS, determine their temporal course up to 90 days post-stroke, and explore their utility as an early diagnostic marker.
Methods And Results:
We used RNA sequencing to study expression changes of circulating microRNAs in a discovery sample of 20 patients with IS and 20 matched healthy control subjects. We further applied quantitative real-time polymerase chain reaction in independent samples for validation (40 patients with IS and 40 matched controls), replication (200 patients with IS, 100 healthy control subjects), and in 72 patients with transient ischemic attacks. Sampling of patient plasma was done immediately upon hospital arrival. We identified, validated, and replicated 3 differentially expressed microRNAs, which were upregulated in patients with IS compared with both healthy control subjects (miR-125a-5p [1.8-fold; P=1.5×10-6], miR-125b-5p [2.5-fold; P=5.6×10-6], and miR-143-3p [4.8-fold; P=7.8×10-9]) and patients with transient ischemic attack (miR-125a-5p: P=0.003; miR-125b-5p: P=0.003; miR-143-3p: P=0.005). Longitudinal analysis of expression levels up to 90 days after stroke revealed a normalization to control levels for miR-125b-5p and miR-143-3p starting at day 2 while miR-125a-5p remained elevated. Levels of all 3 microRNAs depended on platelet numbers in a platelet spike-in experiment but were unaffected by chemical hypoxia in Neuro2a cells and in experimental stroke models. In a random forest classification, miR-125a-5p, miR-125b-5p, and miR-143-3p differentiated between healthy control subjects and patients with IS with an area under the curve of 0.90 (sensitivity: 85.6%; specificity: 76.3%), which was superior to multimodal cranial computed tomography obtained for routine diagnostics (sensitivity: 72.5%) and previously reported biomarkers of acute IS (neuron-specific enolase: area under the curve=0.69; interleukin 6: area under the curve=0.82).
Conclusions:
A set of circulating microRNAs (miR-125a-5p, miR-125b-5p, and miR-143-3p) associates with acute IS and might have clinical utility as an early diagnostic marker.
Insights
Three circulating microRNAs (miRNAs) show potential as early diagnostic biomarkers for acute ischemic stroke (IS). These miRNAs, identified in patient blood, demonstrated high accuracy in distinguishing IS from healthy individuals and transient ischemic attacks.
Area of Science:
- Biochemistry
- Molecular Biology
- Neurology
Background:
- Acute ischemic stroke (IS) lacks clinically useful blood-based biomarkers.
- MicroRNAs (miRNAs) are promising disease markers due to their cell-specific expression and stability in blood.
Purpose of the Study:
- Identify circulating miRNAs associated with acute IS.
- Determine the temporal expression of these miRNAs post-stroke.
- Explore their utility as early diagnostic markers for IS.
Main Methods:
- RNA sequencing and quantitative real-time PCR were used to analyze miRNA expression in discovery, validation, and replication cohorts.
- Plasma samples were collected upon hospital arrival.
- Random forest classification was employed to assess diagnostic performance.
Main Results:
- Three upregulated miRNAs (miR-125a-5p, miR-125b-5p, miR-143-3p) were identified, validated, and replicated in IS patients compared to controls and transient ischemic attack patients.
- These miRNAs showed a diagnostic area under the curve of 0.90, outperforming current methods.
- Expression levels normalized for two miRNAs post-stroke, while one remained elevated.
Conclusions:
- Circulating miR-125a-5p, miR-125b-5p, and miR-143-3p are associated with acute IS.
- These miRNAs hold potential as early diagnostic biomarkers for clinical use in acute ischemic stroke.
