A novel miRNA-4484 is up-regulated on microarray and associated with increased MMP-21 expression in serum of systemic sclerosis patients

Abstract

Systemic sclerosis (SSc) is a complex, heterogeneous connective tissue disease, characterized by fibrosis and ECM deposition in skin and internal organs, autoimmunity, and changes in the microvasculature. Profiling of circulating miRNAs in serum has been found to be changed in pathological states, creating new possibilities for molecular diagnostics as blood-based biomarkers. This study was designed to identify miRNAs that are differentially expressed in SSc and might be potentially contributing to the disease etiopathogenesis or be used for diagnostic purposes. Thus, we compared the expression pattern of multiple miRNAs in serum of 10 SSc patients to 6 healthy controls using microarray analysis, and RT-qPCR to confirm the obtained results. In addition, bioinformatics analysis was performed to explore miRNAs target genes and the signaling pathways that may be potentially involved in SSc pathogenesis. Our study shows a different expression of 15 miRNAs in SSc patients. We identified that miR-4484, located on chromosome 10q26.2, was an 18-fold up-regulated in SSc patients compared to a control group. Bioinformatics analysis of the miR-4484 target genes and the signaling pathways showed that it might be potentially involved in the TGF-β signaling pathway, ECM-receptor interaction, and metalloproteinases expression. Based on the chromosomal location, the most interesting target gene of miR-4484 may be MMP-21. We found that the expression of MMP-21 significantly increased in SSc patients compared to healthy subjects (P < 0.05). Our results suggest that miR-4484, and MMP-21 might be novel serum biomarkers that may correspond to pathological fibrosis in SSc, but it needs to be validated in further studies.

Novel Concepts in Systemic Sclerosis Pathogenesis: Role for miRNAs

“…Matrix metalloproteinase-21 (MMP-21), even though not a direct target gene according to computational analysis, was hypothesized to be up-regulated by miR-4484 due to their close chromosomal vicinity and the increased MMP-21 serum levels. These findings further enabled the authors to suggest that miR-4484 and MMP-21 might play a role in SSc pathogenesis and proposed them as serum biomarkers [45].…”

“…A different miRNA circulating profile was demonstrated by means of microarray analysis in the serum of 10 SSc patients compared to six HC in a study by Rusek et al (2019). Out of the 15 miRNAs differentially expressed, miR-4484 was remarkably increased (18-fold).…”

“…Out of the 15 miRNAs differentially expressed, miR-4484 was remarkably increased (18-fold). Bioinformatics analysis suggested miR-4484 as a potential regulator of fibrosis through the identification of a wide range of target genes involved in the TGF-β/SMAD and Wnt/β-catenin signaling pathways, as well as collagen synthesis and extracellular matrix (ECM) homeostasis [45]. Matrix metalloproteinase-21 (MMP-21), even though not a direct target gene according to computational analysis, was hypothesized to be up-regulated by miR-4484 due to their close chromosomal vicinity and the increased MMP-21 serum levels.…”

“…The subsequent conformational changes in the chromatin architecture make the DNA more or less accessible to transcriptional factors, resulting in activation or repression of gene transcription [42,43]. Non-coding RNAs, miRNAs (<30 nucleotides) [44,45], and lncRNAs (>200 nucleotides) [46,47] are functional regulators of gene expression at the transcriptional and post-transcriptional level. These RNA fragments are transcribed from the DNA but are not translated into proteins [22,23,26,48].…”

Non-Coding RNA in Systemic Sclerosis: A Valuable Tool for Translational and Personalized Medicine

“…Clinical studies show that miRNAs expression profiles in cell-free serum from patients with different autoimmune diseases, including SSc, systemic lupus erythematosus, rheumatoid arthritis, and mixed connective disease [ 140 ]. For instance, Rusek et al have reported an upregulation of novel serum miRNA-4484 associated with increased MMP-21 expression in SSc [ 141 ]. Furthermore, Chouri et al have reported that miRNA-483-5p in serum is a potential driver of fibrosis in SSc [ 142 ].…”