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Published on: February 10, 2015
A liver-specific mouse model for MYO5B-associated cholestasis reveals a toxic gain-of-function as underlying disease
Hui-Yu She1, Yi-Ling Qiu1, Jia-Yan Feng2
1The Center for Pediatric Liver Diseases, Children's Hospital of Fudan University, Shanghai, 201102, China.
Abstract:
Myosin Vb (MYO5B) deficiency, referring to the loss of protein expression or function, causes microvillus inclusion disease (MVID) and/or progressive familial intrahepatic cholestasis-type 10 (PFIC10) in humans. MYO5B plays a role in intracellular trafficking, but the mechanisms by which it contributes to cholestasis are not understood. The aim of this study was to generate a liver-specific mouse model and investigate the mechanism of MYO5B-associated cholestasis. In this study, we generated a liver-specific Myo5b cKO mice via CRISPR/Cas9 genome editing in conjunction with albumin-cre recombinase. Cholestatic stress was induced by dietary-administration of cholic acid (CA) or 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC). To investigate the frequently recurring MYO5B variant (c.2470C > T/p.(Arg824Cys)), adenoviral vectors encoding either the missense variant or blank control sequence were delivered to wild-type and Myo5b cKO mice through tail-vein injection. Serum and liver tissues were harvested from all mice for biochemical and histological analysis. Our findings indicated that loss of Myo5b expression did not cause cholestatic liver disease and did not augment CA or DDC feeding-induced cholestatic stress. By contrast, expression of the MYO5B c.2470C > T/p. (Arg824Cys) variant induced cholestasis, evidenced by elevated levels of serum alanine aminotransferase, alkaline phosphatase and bilirubin, mild hepatocellular injury, and altered bile salt export pump (Bsep) localization, resembling that observed in human PFIC10. In summary, we have developed a mouse model of MYO5B-associated cholestasis. The expression of the MYO5B-p. (Arg824Cys) variant but not the loss of Myo5b expression caused cholestasis, indicating a toxic gain-of-function as underlying disease mechanism.
Insights
A specific MYO5B variant, not its absence, causes cholestasis by altering bile salt export. This study developed a mouse model to investigate this toxic gain-of-function mechanism in liver disease.
Area of Science:
- Hepatology
- Molecular Biology
- Genetics
Background:
- Myosin Vb (MYO5B) deficiency is linked to human liver diseases like progressive familial intrahepatic cholestasis-type 10 (PFIC10).
- The precise mechanisms by which MYO5B dysfunction leads to cholestasis remain unclear.
- Understanding MYO5B's role is crucial for deciphering cholestatic liver disease pathogenesis.
Purpose of the Study:
- To generate a liver-specific mouse model for investigating MYO5B-associated cholestasis.
- To elucidate the molecular mechanisms underlying MYO5B-related liver disease.
- To differentiate between loss-of-function and potential toxic gain-of-function of MYO5B variants.
Main Methods:
- Generated liver-specific Myo5b conditional knockout (cKO) mice using CRISPR/Cas9 and albumin-cre.
- Induced cholestatic stress via dietary cholic acid (CA) or 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) feeding.
- Utilized adenoviral vectors to express a common MYO5B variant (c.2470C>T/p.Arg824Cys) in wild-type and cKO mice.
Main Results:
- Loss of Myo5b expression in mice did not induce cholestasis or exacerbate chemically induced liver injury.
- Expression of the MYO5B p.Arg824Cys variant triggered cholestasis, characterized by elevated liver enzymes and bilirubin.
- The MYO5B variant led to altered bile salt export pump (Bsep) localization, mimicking human PFIC10 phenotypes.
Conclusions:
- A novel mouse model for MYO5B-associated cholestasis was successfully developed.
- Cholestasis resulted from the expression of the specific MYO5B variant, not MYO5B deficiency.
- Findings suggest a toxic gain-of-function mechanism for MYO5B-related cholestatic liver disease.
