MicroRNA-223 Suppresses Human Hepatic Stellate Cell Activation Partly via Regulating the Actin Cytoskeleton and Alleviates Fibrosis in Organoid Models of Liver Injury

Abstract

MicroRNAs (miRNAs) are small, non-coding RNAs that negatively regulate target mRNA expression, and altered expression of miRNAs is associated with liver pathological conditions. Recent studies in animal models have shown neutrophil/myeloid-specific microRNA-223 (miR-223) as a key regulator in the development of various liver diseases including fibrosis, where hepatic stellate cells (HSCs) are the key player in pathogenesis. However, the precise roles of miR-223 in human HSCs and its therapeutic potential to control fibrosis remain largely unexplored. Using primary human HSCs, we demonstrated that miR-223 suppressed the fibrogenic program and cellular proliferation while promoting features of quiescent HSCs including lipid re-accumulation and retinol storage. Furthermore, induction of miR-223 in HSCs decreased cellular motility and contraction. Mechanistically, miR-223 negatively regulated expression of smooth muscle α-actin (α-SMA) and thus reduced cytoskeletal activity, which is known to promote amplification of fibrogenic signals. Restoration of α-SMA in miR-223-overexpressing HSCs alleviated the antifibrotic effects of miR-223. Finally, to explore the therapeutic potential of miR-233 in liver fibrosis, we generated co-cultured organoids of HSCs with Huh7 hepatoma cells and challenged them with acetaminophen (APAP) or palmitic acid (PA) to induce hepatotoxicity. We showed that ectopic expression of miR-223 in HSCs attenuated fibrogenesis in the two human organoid models of liver injury, suggesting its potential application in antifibrotic therapy.

Long non-coding RNA H19 promotes proliferation in hepatocellular carcinoma cells via H19/miR-107/CDK6 axis

“…Annealed oligonucleotides of H19-specific and control shRNA sequences were cloned into a lentiviral vector, Tet-LKO-Puro (AddGene Plasmid #21915), and confirmed for proper insertion by Sanger sequencing. Lentivirus was produced in HEK293FT cells and used to establish Doxinducible shRNA cell lines as previously described [23]. To suppress the activity of microRNA-107 (miR-107), we cloned a sequence of antagomiR against miR-107 into Tet-LKO-Puro and established transgenic HCC lines with Doxinducible expression of antagomiR.…”

“…Transfected cells were employed for subsequent analysis after 48 h posttransfection. For overexpression of miR-107, the pre-miRNA sequence of miR-107 was cloned into a lentiviral vector as previously described [23]. Primers used for cloning can be found in Suppl.…”

The diagnostic performance of microRNAs expression levels in cellular aspirate of solitary thyroid nodule overrides that of cytology

“…1 PTC is the most common histological subtype among the recently diagnosed TC patients, accounts for 90% of all cases, and is microRNAs expression is associated with deregulation of the target RNA expression with subsequent altered function of the resultant protein and development of pathological conditions of the affected organs. 4 The role of multiple microRNAs in pathogenesis of TC was recently declared; lower expression levels of miR-30b was found to promote TC via induction of high expression levels of ubiquitin-specific protease-22 in thyroid cells and downregulation of miR-27a could facilitate the development of PTC through promoting the expression of collagen, and calcium-binding EGF domain-containing protein-1 in thyroid tissue. 5,6 On contrary, upregulated Mir-449a suppresses PTC development through binding to the 3' un-translated region of Metadherin and downregulating its expression.…”

Prophylactic Resveratrol Ameliorates Thioacetamide Hepatotoxicity in A Dose-Dependent Fashion Through the Regulation of Gene Expression Levels of MiR-155 and MiR-21

“…MicroRNAs (MiR-) are small, non-coding RNAs, which regulate the expression of the target mRNA, and alteration of MiR expressions is associated with pathological conditions of the affected organs (Ariyachet et al,2022). Specific microRNAs was suggested to affect the HSCs functions; MiR-378 and MiR-17 play a role in the pathogenesis of LF through activation of HSCs via altering TGF-β/smads and Wnt/β-catenin pathways (Zaafan and Abdelhamid.,2022).…”

“…Specific microRNAs was suggested to affect the HSCs functions; MiR-378 and MiR-17 play a role in the pathogenesis of LF through activation of HSCs via altering TGF-β/smads and Wnt/β-catenin pathways (Zaafan and Abdelhamid.,2022). Also, neutrophil/myeloid-specific miR-223 has a key regulator role in the development of LF through the activation of HSCs (Ariyachet et al,2022). However, miR-122 suppresses liver cirrhosis through its inhibitory effect on HSCs by targeting the EphB2 expression (Ma et al,2022).…”