Inhibition of CUB and sushi multiple domains 1 (CSMD1) expression by miRNA-190a-3p enhances hypertrophic scar-derived fibroblast migration in vitro

Abstract

Background Hypertrophic scar (HTS) is a fibroproliferative skin disorder characterized by excessive cell proliferation, migration, and extracellular matrix (ECM) deposition. The CUB and Sushi multiple domains 1 (CSMD1) has previously been identified as the key regulatory gene of hypertrophic scar by a large sample GWAS study. However, further research has not yet been conducted to verify this finding in other HTS patients and to determine the underlying mechanism. Results In this study, we verified that CSMD1 was downregulated in both HTS tissue and HTS-derived fibroblasts. The knockdown of CSMD1 resulted in enhanced migration and fibronectin1 (FN1) secretion in fibroblasts in vitro. In addition, the upstream and downstream regulatory mechanisms of CSMD1 were also investigated through microRNA (miRNA) databases screening and RNA-sequencing (RNA-seq) respectively. The screening of four common microRNA (miRNA) databases suggested that miR-190a-3p binds to the CSMD1 and may regulate its expression. We confirmed that miR-190a-3p directly targeted the CSMD1–3′-UTR using luciferase reporter assays. Furthermore, the overexpression of miR-190a-3p showed promotion of migratory activity and FN1 secretion in fibroblasts, resembling the effect of CSMD1 knockdown; whereas the knockdown of miR-190a-3p exerted the opposite effect. Finally, transcriptomic analysis showed activation of Janus kinase-signal transducer and activator of transcription (JAK/STAT) signaling pathway in the CSMD1 knockdown fibroblasts. Conclusions This study has validated the conclusions of the previous GWAS study conducted in Chinese population. In vitro experiments have provided further evidence on the function of CSMD1 in the development of HTS, and have also revealed the underlying upstream and downstream regulating mechanisms. Additionally, the JAK/STAT signaling pathway identified using RNA-seq might provide a potential treatment approach, especially for HTS.

Exosome Derived from Mesenchymal Stem Cells Alleviates Pathological Scars by Inhibiting the Proliferation, Migration and Protein Expression of Fibroblasts via Delivering miR-138-5p to Target SIRT1

“…Moreover, therapies using miRNAs are promising means of regulating the gene expression involved in wound healing and scar formation. [56][57][58] miRNAs could also exert their work as a modification to MSCs or MSC-Exo, reducing excessive scar formation. As previous studies introduced, the MSCs or MSC-Exo modified by miR-181-5p, miR-29a, miR-146a, miR-21, miR-146a-5p, etc.…”

miR-361 prevents the formation of hypertrophic scars by inhibiting TGF-β1

“…miR-3613-3p has been shown to inhibit HS formation by targeting arginine and glutamate-rich 1, which may provide potential therapeutic targets for the management of HS [16] . miR-190a-3p in HS can bind to the CUB and sushi multiple domains 1 and may regulate its expression, thus affecting the occurrence and development of HS [19] ). miR-145-5p can prevent ber formation and reduce the formation of HS by reducing the expression of Smad2/3 [18] .…”

HSFAS mediates fibroblast proliferation, migration, trans-differentiation and apoptosis in hypertrophic scars via interacting with ADAMTS8

“…We further confirmed its expression and sequence in fibroblasts by Northern blot and RACE assays. Accumulating evidence suggests that the pathogenesis of HS is involved in the abnormally increased proliferation and migration of fibroblasts, thereby leading to apoptosis inhibition [ 29 , 30 ]. Moreover, the trans-differentiation of fibroblasts to myofibroblasts is also a critical procedure in the pathogenesis of scar formation, which is characterized by alpha smooth muscle actin-positive (α-SMA + ) fibroblasts that can stimulate collagen synthesis, particularly Col I and Col III [ 31 ‒ 33 ].…”