RSPO4-CRISPR alleviates liver injury and restores gut microbiota in a rat model of liver fibrosis

Linghua Yu,Huixing Yi,Linlin Wang,Xiaojun Wu

doi:10.1038/s42003-021-01747-5

Abstract

Wnt signaling dysfunction and gut dysbiosis may lead to liver fibrosis, yet the underlying mechanisms are not well elucidated. This study demonstrated the role of RSPO4, a Wnt signaling agonist, in liver fibrogenesis and its impact on the gut microbiome. RSPO4 gene in CCl4-induced fibrotic-liver rats was knockout by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system, with healthy rats served as the control. Tissue samples and hepatic stellate cells (HSCs) isolated from rats were examined for curative effect of RSPO4-CRISPR treatment. Fecal sample were collected and analyzed with 16 S rRNA sequencing. We found RSPO4-CRISPR relieved liver fibrosis in rats and reversed HSC activation. Further, results showed RSPO4-CRISPR tended to restore the microflora composition. Significance species between groups were identified. Bacteroides and Escherichia-Shigella were the key microbes in the model and negative group, whereas Lactobacillus, Romboutsia, and Lachnospiraceae NK4A136 group were abundant in the control. Notably, Bacteroidales S24-7 group and Ruminococcaceae UCG-005 were the significantly enriched in CRISPR group. We show that the microbiome of rats treated with RSPO4-CRISPR presents a trend towards the restoration of the original condition. Our findings pave a new way to evaluate the curative effect of liver fibrosis treatment.

Highlights

Wnt signaling dysfunction and gut dysbiosis may lead to liver fibrosis, yet the underlying mechanisms are not well elucidated
Masson staining of the rat liver tissues revealed that Roof platespecific spondin 4 (RSPO4)-Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) may slow down liver fibrogenesis (Fig. 1A)
These findings suggest that RSPO4-CRISPR may relieve liver fibrogenesis in rats

Summary

Introduction

Wnt signaling dysfunction and gut dysbiosis may lead to liver fibrosis, yet the underlying mechanisms are not well elucidated. This study demonstrated the role of RSPO4, a Wnt signaling agonist, in liver fibrogenesis and its impact on the gut microbiome. RSPO4 gene in CCl4-induced fibrotic-liver rats was knockout by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system, with healthy rats served as the control. Tissue samples and hepatic stellate cells (HSCs) isolated from rats were examined for curative effect of RSPO4-CRISPR treatment. We knocked out the RSPO4 gene in rats with CCl4-induced liver fibrosis using the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system. This study aimed to identify a panel of significant gut microbial species associated with liver fibrosis and its regression to normal status by controlling the underlying causes, which could be used to evaluate the therapeutic effectiveness of liver fibrosis treatments

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