Long non-coding RNA crnde promotes deep vein thrombosis by sequestering miR-181a-5p away from thrombogenic Pcyox1l

Abstract

Background Deep vein thrombosis (DVT) is an interplay of genetic and acquired risk factors, where functional interactions in lncRNA-miRNA-mRNA ceRNA networks contribute to disease pathogenesis. Based on the high-throughput transcriptome sequencing prediction, we have assessed the contribution of lncRNA Crnde/miR-181a-5p/Pcyox1l axis to thrombus formation. Methods DVT was modeled in mice by inferior vena cava stenosis, and inferior vena cava tissues were harvested for high-throughput transcriptome sequencing to screen differentially expressed lncRNAs and mRNAs. The key miRNA binding to Crnde and Pcyox1l was obtained through searching the RNAInter and mirWalk databases. The binding affinity between Crnde, miR-181a-5p, and Pcyox1l was examined by FISH, dual luciferase reporter gene, RNA pull-down, and RIP assays. Functional experiments were conducted in DVT mouse models to assess thrombus formation and inflammatory injury in inferior vena cava. Results It was noted that Crnde and Pcyox1l were upregulated in the blood of DVT mice. Crnde competitively bound to miR-181a-5p and inhibited miR-181a-5p expression, and Pcyox1l was the downstream target gene of miR-181a-5p. Silencing of Crnde or restoration of miR-181a-5p reduced inflammatory injury in the inferior vena cava, thus curtailing thrombus formation in mice. Ectopic expression of Pcyox1l counterweighed the inhibitory effect of Crnde silencing. Conclusions Therefore, Crnde sequesters miR-181a-5p to release Pcyox1l expression via ceRNA mechanism, thus aggravating thrombus formation in DVT.

Long Non-Coding RNAs in Venous Thromboembolism: Where Do We Stand?

“…More recent studies have indicated that this lncRNA is also implicated in other malignancies, in addition to regulating vascular smooth muscle cell behaviour and inflammation and apoptosis in alcoholic liver disease [125][126][127][128]. A recent study found that CRNDE and the prenylcysteine oxidase 1 (PCYOX1) were upregulated in the blood of DVT mice [129]. The protein PCYOX1 is an enzyme involved in the breakdown of prenylated proteins found in vascular and blood cells.…”

“…Deficiency of PCYOX1 is suggested to cause platelet hypo-reactivity [130,131]. Thus, acting as a decoy and through the sequestering of miR-181a-5p (targets PCYOX1), CRNDE might lead to PCYOX1 upregulation, exacerbating thrombus formation and vascular inflammatory injury in DVT [129].…”

MiRNAs and Their Role in Venous Thromboembolic Complications

“…In other words, the aberrant expression of miR-195 and miR-582 can cause endothelial dysfunction by regulating vascular homeostasis and can ultimately lead to thrombus formation in both arteries and veins. Table 2 summarizes the results of other studies that assessed the role of some representative miRNAs in DVT pathogenesis [46][47][48][49][50][51][52][53][54][55][56][57][58][59][60][61].…”