Effective delivery of osteopontin small interference RNA using exosomes suppresses liver fibrosis via TGF-β1 signaling

Abstract

Objective and aims: Osteopontin (OPN), an oxidant stress sensitive cytokine, plays a central role in liver fibrosis. While OPN expression can be reduced by small interfering RNA (siRNA), the challenge to deliver siRNA safely and effectively into liver remains unresolved. Exosomes are promising natural nanocarriers for drug delivery that are able to enter cells with different biological barriers efficiently. In this study, we used exosomes as a delivery vehicle to target OPN in liver fibrosis.Methods: Exosomes selectively home to fibrotic liver according to small animal imaging system. Electroporation technique was used to engineer exosomes to carry siRNA targeting OPN (ExosiRNA−OPN). Primary hepatic stellate cells (HSCs) were isolated and treated with ExosiRNA−OPN to assess the effect on activated HSCs (aHSCs). Immunofluorescence for α−SMA, an aHSCs marker, and sirius red staining were performed to assess ECM deposition. Finally, plasma OPN from patients with liver fibrosis was identified by ELISA assay.Results: Exosome-mediated siRNA delivery systems show high uptake and low toxicity. Besides, ExosiRNA−OPN suppressed HSCs activation and ECM deposition and more efficiently improved liver function when compared to naked siRNA-OPN. Moreover, ExosiRNA−OPN was assumed inhibiting TGF-β1 signaling activation, along with other fibrotic-related genes based on a GEO datasheet of liver fibrosis samples for correlation analyzes. ExosiRNA−OPN inhibited TGF-β1 signaling by decreasing high-mobility group box-1 (HMGB1). Plasma proteins from chronic HBV-induced fibrosis patients were identified that patients with high OPN expression correlates with more advanced fibrosis progression.Discussion: This study shows that exosome-mediated siRNA-OPN delivery may be an effective option for the treatment of liver fibrosis.

Potential of PAQosome as a therapeutic target for hepatic fibrosis

“…This indicated that ASDURF may be a regulator of TGF-β1/Smad3 and NF-κB signaling pathways, and TGF-β1/Smad3 and NF-κB are important pathways in hepatic fibrosis. [3][4][5] This further supports the hypothesis that ASDURF is involved in managing hepatic fibrosis. The dual-luciferase test revealed that ASDURF enhanced ASNSD1 promoter activity, and Co-IP demonstrated that ASDURF could attach to ASNSD1 (Fig.…”

“…1 The most traditional method for activating HSCs is through the transforming growth factor-β1 (TGFβ1)/ Sekelsky mothers against decapentaplegic homologue 3 (Smad3) signaling pathway. 2,3 Additionally, NF-κB controls hepatic fibrogenesis. 4,5 Furthermore, the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway is linked to HSC activation, proliferation, and extracellular matrix production according to a number of studies.…”

Liver fibrosis: pathological features, clinical treatment and application of therapeutic nanoagents

“…The Exo siRNA-OPN selectively homed to the fibrotic liver and inhibited the TGF-b1 signaling pathway by reducing HMGB1, inhibited HSC activation and ECM deposition, and more effectively ameliorated liver function. 158 Although EVs-based antifibrotic therapy is a promising option, its application still has certain limitations. The heterogeneity of EV types and the presence of other bionanoparticles with overlapping properties make their yields low, formulations inconsistent, and isolation and acquisition challenging.…”

“…The Exo siRNA-OPN selectively homed to the fibrotic liver and inhibited the TGF-β1 signaling pathway by reducing HMGB1, inhibited HSC activation and ECM deposition, and more effectively ameliorated liver function. 158…”

Exosomes in liver fibrosis: The role of modulating hepatic stellate cells and immune cells, and prospects for clinical applications

“…used electroporation to enable exosomes to carry siRNA targeting osteopontin to inhibit LF. This exosome delivery system showed high uptake and low toxicity, effectively improving the low delivery efficiency of siRNA to target organs ( 103 ). In addition, He et al.…”