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Bile Duct Ligation in Mice: Induction of Inflammatory Liver Injury and Fibrosis by Obstructive Cholestasis
Published on: February 10, 2015
Targeted Apoptosis of Ductular Reactive Cells Reduces Hepatic Fibrosis in a Mouse Model of Cholestasis
Adiba I Azad1, Anuradha Krishnan1, Leia Troop1
1Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN.
Background And Aims:
In cholestatic liver diseases, ductular reactive (DR) cells extend into the hepatic parenchyma and promote inflammation and fibrosis. We have previously observed that multidrug-resistant 2 (Mdr2-/- ) double knockout (DKO) mice lacking tumor necrosis factor-related apoptosis-inducing ligand receptor (Tr-/- ) display a more extensive ductular reaction and hepatic fibrosis compared to Mdr2-/- mice. This observation suggests that the magnitude of the DR-cell population may be regulated by apoptosis.
Approach And Results:
To examine this concept, we cultured epithelial cell adhesion molecule-positive reactive cholangioids (ERCs) obtained from wild-type (WT), Tr-/- , Mdr2-/- and DKO mice. Single-cell transcriptomics and immunostaining of both WT and DKO ERCs confirmed their DR-cell phenotype. Moreover, DKO ERCs displayed a unique translational cluster with expression of chemokines, indicating a reactive state. Incubation with the myeloid cell leukemia 1 (MCL1) inhibitor S63845, a proapoptotic BH3-mimetic therapy, significantly decreased DKO and Mdr2-/- ERC viability compared to WT. Intravenous administration of S63845 significantly reduced the DR-cell population and markers of inflammation and liver fibrosis in Mdr2-/- and DKO mice. Furthermore, DKO mice treated with S63845 displayed a significant decrease in hepatic B lymphocytes compared to untreated mice as assessed by high-definition mass cytometry by time-of-flight. Coculture of bone marrow-derived macrophages with ERCs from DKO mouse livers up-regulated expression of the B cell-directed chemokine (C-C motif) ligand 5. Finally, DR cells were noted to be primed for apoptosis with Bcl-2 homologous antagonist/killer activation in vitro and in vivo in primary sclerosing cholangitis liver specimens.
Conclusions:
DR cells appear to play a key role in recruiting immune cells to the liver to actively create an inflammatory and profibrogenic microenvironment. Pharmacologic targeting of MCL1 in a mouse model of chronic cholestasis reduces DR-cell and B-cell populations and hepatic fibrosis.
Insights
Targeting myeloid cell leukemia 1 (MCL1) with S63845 reduces ductular reactive (DR) cells and liver fibrosis in cholestatic liver disease models. This proapoptotic therapy decreases DR-cell populations and associated inflammation and B-cell recruitment.
Area of Science:
- Hepatology
- Cell Biology
- Immunology
Background:
- Ductular reactive (DR) cells in cholestatic liver diseases promote inflammation and fibrosis.
- The magnitude of DR-cell populations may be regulated by apoptosis.
- Previous observations in Mdr2 knockout mice suggest a link between apoptosis and DR-cell extent.
Purpose of the Study:
- To investigate the role of apoptosis in regulating DR-cell populations.
- To examine the therapeutic potential of targeting MCL1 (myeloid cell leukemia 1) in a mouse model of chronic cholestasis.
Main Methods:
- Primary reactive cholangioid (ERC) culture from wild-type and knockout mice.
- Single-cell transcriptomics and immunostaining to characterize ERCs.
- In vitro and in vivo treatment with MCL1 inhibitor S63845.
- Assessment of DR-cell populations, inflammation, fibrosis, and immune cell infiltration.
Main Results:
- DKO ERCs showed a unique reactive state with chemokine expression.
- S63845 significantly decreased ERC viability and DR-cell populations in Mdr2-/- and DKO mice.
- S63845 treatment reduced liver inflammation, fibrosis, and hepatic B lymphocytes.
- DR cells were primed for apoptosis in vitro and in vivo, including in primary sclerosing cholangitis specimens.
Conclusions:
- DR cells contribute to an inflammatory and profibrogenic liver microenvironment by recruiting immune cells.
- Pharmacologic targeting of MCL1 effectively reduces DR-cell and B-cell populations in a cholestasis model.
- MCL1 inhibition represents a potential therapeutic strategy for chronic cholestatic liver diseases.
