Related Experiment Video
Updated: May 21, 2026
A Mouse Model of Chronic Liver Fibrosis for the Study of Biliary Atresia
Published on: February 3, 2023
Single-cell transcriptomics reveals anti-inflammatory effects of prednisolone on the hepatic niche in biliary atresia
Paul D Blakeley1, Fangran Liu1, Zhongluan Wu1
1Department of Surgery, Li Ka Shing Faculty of Medicine, School of Clinical Medicine, Queen Mary Hospital, The University of Hong Kong, 102 Pokfulam Road, Hong Kong SAR, China.
Insights
Prednisolone treatment ameliorates biliary atresia (BA) by modulating liver cells. It suppresses inflammation and promotes cell quiescence, offering a potential therapeutic strategy for this pediatric liver disease.
Area of Science:
- Hepatology
- Immunology
- Molecular Biology
Background:
- Biliary atresia (BA) is a severe pediatric liver disease causing bile duct obstruction and fibrosis.
- Current treatments for BA are limited, often requiring surgery or transplantation.
- The effectiveness and mechanism of prednisolone in BA treatment are not well-established.
Purpose of the Study:
- To investigate the therapeutic effects and underlying mechanisms of prednisolone in a Rhesus Rotavirus (RRV)-induced mouse model of biliary atresia.
- To identify specific cell populations and gene regulatory networks (GRNs) involved in prednisolone's action.
Main Methods:
- Induction of BA in mice using Rhesus Rotavirus (RRV).
- Administration of prednisolone or placebo (PBS) to infected mice.
- Single-cell RNA sequencing (scRNA-seq) and immunohistochemistry on liver tissues.
- Computational reconstruction of gene regulatory networks.
Main Results:
- Prednisolone treatment reduced pro-inflammatory macrophages and increased immunosuppressive Kupffer cells.
- Prednisolone promoted the deactivation of hepatic stellate cells (HSCs) into a quiescent state.
- A pro-inflammatory myofibroblast subpopulation (MFB-2), driven by the Fosl1-Hnf1b GRN, was identified and showed resistance to prednisolone.
- HSC and MyoFb subtypes were conserved in BA patient livers.
Conclusions:
- Prednisolone transforms the liver immune-mesenchymal niche from inflammatory to regenerative in BA.
- Modulation of GRNs in HSCs is crucial for prednisolone's therapeutic effect in BA.
- Targeting specific HSC and myofibroblast subpopulations may offer novel therapeutic strategies for biliary atresia.
Abstract:
Biliary atresia (BA) is a life-threatening paediatric liver disease, characterized by bile duct obstruction, progressive inflammation and fibrosis. Treatment options are limited, often necessitating surgery or liver transplantation to prevent irreversible liver damage. Prednisolone has shown promise in treating BA; however, there is currently no consensus on its effectiveness, and the mechanism of action remains unknown. We investigated the effects of prednisolone in a BA model induced by Rhesus Rotavirus (RRV). Liver samples were collected from day-21 and day-28 post-RRV infected mice injected with PBS or steroid, and subjected to single-cell RNA sequencing (scRNA-seq) and immunohistochemistry. We identified subpopulations of hepatic stellate cells (HSCs) and macrophages that serve as key effectors of prednisolone-mediated disease amelioration. Prednisolone treatment at 28-days post RRV-infection suppressed pro-inflammatory Ly6c-high macrophages, increased immunosuppressive M2 Kupffer cells, and promoted deactivation of HSCs into the quiescent state. Computational reconstruction of HSC differentiation pathways revealed that the Nr3c1-Klf7 gene regulatory network (GRN) modulates the transition of HSCs between quiescence and activation, and we show that HSCs differentiate into three myofibroblast (MyoFbs) subpopulations, each exhibiting different pathogenic potential. We identified a pro-inflammatory MyoFB subpopulation (MFB-2) characterized by the Fosl1-Hnf1b GRN. MFB-2 secretes elevated levels of macrophage-activating cytokines, exhibits resistance to apoptotic signals, and displayed impaired responsiveness to prednisolone. The HSC and MyoFb subtypes were also identified in BA patient livers with highly conserved gene expression and functional profiles, supporting their importance in BA progression. Prednisolone treatment transforms the liver immune-mesenchymal niche from an inflammatory to regenerative state in BA and modulation of GRNs in HSCs plays a pivotal role.
