Screening of Acute Myocardial Infarction Related piRNA in Plasma Exosomal Based on Massively Parallel Sequencing

Abstract:
Objective: This study aims to explore plasma exosomal piRNAs (Piwi-interacting RNAs) associated with acute myocardial infarction (AMI) progression, particularly those capable of interacting with PIWI proteins. Furthermore, we preliminarily investigate the correlation between these piRNAs and AMI to evaluate their potential as diagnostic biomarkers for AMI. Methods: A total of 12 male Wistar rats aged 6-8 weeks, weighing 210-310 g, were selected to establish AMI models with varying infarction time points by ligating the left anterior descending branch of the coronary artery (LAD). The rats were divided into two groups: Sham group and AMI group, with 6 rats in each group. Blood samples were subsequently collected from each group, and plasma was isolated. Exosomes and their contained RNA were further extracted from the plasma. We employed massively parallel sequencing (MPS) technology to compare the two groups and validated the differentially expressed piRNAs in rat plasma exosomes by real-time quantitative PCR (qPCR). Based on the piRBase database, further target gene analysis of the identified piRNAs was conducted. Results: MPS detected a total of 39,930 piRNAs. Compared to the Sham group, the expression profiles of piRNAs in the AMI group showed significant changes, with 15,777 piRNAs upregulated and 9,695 piRNAs downregulated. Validation by qPCR confirmed that the expression changes of piR-rno-1287165, piR-rno-568508, piR-rno-2058103, piR-rno-2375534, piR-rno-863759, and piR-rno-2096428 were consistent with the sequencing results, showing increased expression levels following AMI. Target gene analysis of the differentially expressed piRNAs was performed. The results revealed that piR-rno-568508 has 2 target genes (Wwp, Mir140), piR-rno-2096428 has 2 target genes (Gene symbol: AABR07015057.1, AABR07015081.2), and piR-rno-2058103 has 3 target genes (Gene symbol: AABR07015067.1, AABR07015081.2, AABR07063425.3). Conclusion: During AMI progression, six plasma exosomal piRNAs (piR-rno-1287165, piR-rno-568508, piR-rno- 2058103, piR-rno-2375534, piR-rno-863759, and piR-rno-2096428) showed significant expression changes, suggesting their potential as biomarkers for early differential diagnosis of AMI. The identification and functional analysis of these piRNA target genes offer valuable insights for the clinical diagnosis, treatment, and prognosis of AMI. Moreover, this finding provides a novel approach for early differential diagnosis of AMI in forensic medicine, advancing diagnostic technologies in this field.