Plasma-Derived Exosomal hsa-miR-4488 and hsa-miR-1228-5p: Novel Biomarkers for Dermatomyositis-Associated Interstitial Lung Disease with Anti-Melanoma Differentiation-Associated Protein 5 Antibody-Positive Subset

Abstract

The present study is aimed at profiling circulating exosome-derived microRNAs (miRNAs/miRs) from patients with dermatomyositis (DM), in particular those complicated with interstitial lung disease (ILD) with anti-melanoma differentiation-associated protein 5 (MDA5) antibody-positive. Fifteen participants were enrolled, including five patients with DM complicated with ILDs prior to treatment with circulating anti-MDA5 antibody-positive status [DM-ILD-MDA5 Ab(+)], five DM patients without ILDs who were negative for 16 detectable myositis-specific antibodies [DM-nonILD-MSA16(-)], and five age- and gender-matched healthy donor controls (HCs). The characteristics of the exosomes extracted by Ribo™ Exosome Isolation Reagent were identified using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and flow cytometry. Differentially expressed miRNAs, determined by next-generation deep sequencing, were identified through the criteria of ∣ log 2 fold change ∣ ≥ 1 and P < 0.01 . A total of 38 miRNAs were significantly upregulated in exosomes from patients with DM-ILD-MDA5 Ab(+) compared to those from HC, while 21 miRNAs were significantly downregulated. Compared to exosomes derived from patients with DM-nonILD-MSA16(-), 51 miRNAs were significantly upregulated and 33 miRNAs were significantly downregulated from patients with DM-ILD-MDA5 Ab(+). A total of 73 exosomal miRNAs were significantly differentially expressed between DM-nonILD-MSA16(-) and HC. In particular, two miRNAs, Homo sapiens- (hsa-) miR-4488 and hsa-miR-1228-5p, were common differentially expressed miRNAs among three comparisons. GO and KEGG analyses suggested that several pathways may contribute the pathogenesis of DM-ILD-MDA5 Ab(+) and DM-nonILD-MSA16(-), while PPI network analysis of hsa-miR-4488 and hsa-miR-1228-5p indicated that their predicted target genes, DExD-box helicase 39B and MDM2, may be involved in the mechanisms of DM-ILD-MDA5 Ab(+).

The Clinical Significance of Transfer RNAs Present in Extracellular Vesicles

“…However, tRF-28-QSZ34KRQ590K did not show any significant correlation with the disease severity of atopic dermatitis. Another autoimmune condition was examined by Zhong et al [ 74 ], where the exosomal RNA profile from the blood samples of 10 dermatomyositis patients, with half of the patients having simultaneous interstitial lung disease (ILD) involvement, was investigated. In all three patient groups, namely dermatomyositis patients with ILD and without ILD and control patients, exosomal tRNA represented one of the smallest proportions of sRNA, with 0.42%, 0.49% and 0.64%, respectively.…”

Hyperoxia Induced Bronchopulmonary Dysplasia-Like Inflammation via miR34a-TNIP2-IL-1β Pathway

“…Therefore, biomarkers or therapeutic targets in BPD diagnostics and prediction appear to be particularly important ( 40 ). MiRNAs have been extremely studied in many diseases due to their utility in diagnostics of oncology ( 41 ), infectious diseases ( 42 ), and lung disease ( 43 ). Previous research found that miR34a was considerably increased in the lungs of mice subjected to hyperoxia compared to normal controls, but decreased when exposed to hypoxia ( 35 , 44 ).…”

Roles of biomarkers in anti‐MDA5‐positive dermatomyositis, associated interstitial lung disease, and rapidly progressive interstitial lung disease

“…Exosomal microRNAs (miRNAs) stably circulate in the blood and are biomarkers for various diseases. A study showed that upregulated plasma‐derived exosomal miR‐4488 and miR‐1228‐5p might help diagnose, prevent, and treat MDA5 + DM‐ILD based on next‐generation sequencing 96 . Besides miRNA, high serum levels of cell‐free DNA (cfDNA), mainly produced by neutrophil extracellular traps (NETs), may reflect NETs involved in MDA5 + DM‐ILD pathogenesis 97 .…”

“…A study showed that upregulated plasma‐derived exosomal miR‐4488 and miR‐1228‐5p might help diagnose, prevent, and treat MDA5 + DM‐ILD based on next‐generation sequencing. 96 Besides miRNA, high serum levels of cell‐free DNA (cfDNA), mainly produced by neutrophil extracellular traps (NETs), may reflect NETs involved in MDA5 + DM‐ILD pathogenesis. 97 Circulating NET [both human neutrophil elastase (HNE)‐DNA and myeloperoxidase (MPO)‐DNA complexes] are elevated in MDA5 + DM patients, suggesting that anti‐MDA5 antibodies may enhance NET formation, reflecting neutrophil dysregulation.…”