Intestinal dysbiosis drives liver disease progression via NLRP3 in the Mdr2-/- model of primary sclerosing cholangitis

KM Schneider; L Liao; EJC Galvez; M Frissen; HU Marschall; H Su; M Hatting; A Wahlström; J Haybaeck; P Puchas; A Mohs; J Peng; J Jung; J Reißing; HW Zimmermann; T Longerich; T Strowig; C Liedtke; FJ Cubero; C Trautwein

doi:10.1055/s-0038-1677100

Zeitschrift für Gastroenterologie, Table of Contents

Z Gastroenterol 2019; 57(01): e23
DOI: 10.1055/s-0038-1677100

1. Basic Hepatology (Fibrogenesis, NPC, Transport)

Georg Thieme Verlag KG Stuttgart · New York

Intestinal dysbiosis drives liver disease progression via NLRP3 in the Mdr2-/- model of primary sclerosing cholangitis

KM Schneider

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

L Liao

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

²Department of Anesthesiology and Pain Management, Shanghai East Hospital, Tongji University, Shanghai, China.

,

EJC Galvez

³Helmholtz Centre for Infection Research, Braunschweig, Germany.

,

M Frissen

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

HU Marschall

⁴Wallenberg Laboratory, Department of Molecular and Clinical Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.

,

H Su

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

M Hatting

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

A Wahlström

⁴Wallenberg Laboratory, Department of Molecular and Clinical Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.

,

J Haybaeck

⁵Department of Pathology, Otto von Guericke University Magdeburg, Magdeburg, Germany.

⁶Institute of Pathology, Medical University Graz, Graz, Austria.

⁷Department of Pathology, Medical University Innsbruck, Innsbruck, Austria.

,

P Puchas

⁶Institute of Pathology, Medical University Graz, Graz, Austria.

,

A Mohs

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

J Peng

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

J Jung

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

J Reißing

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

HW Zimmermann

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

T Longerich

⁸Liver Cancer Center Heidelberg, Heidelberg University Hospital, INF 224, 69120, Heidelberg, Germany.

⁹Division Translational Gastrointestinal Pathology, Institute of Pathology, Heidelberg University Hospital, INF 224, 69120, Heidelberg, Germany.

,

T Strowig

³Helmholtz Centre for Infection Research, Braunschweig, Germany.

,

C Liedtke

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

,

FJ Cubero

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

¹⁰Department of Immunology, Complutense University School of Medicine, and 12 de Octubre Health Research Institute (imas12), Madrid 28041, Spain.

,

C Trautwein

¹Department of Medicine III, University Hospital RWTH Aachen, Germany.

› Author Affiliations

Abstract

Full Text

Background:

There is a striking association between human cholestatic liver disease and inflammatory bowel disease. However, the implications for intestinal microbiota and inflammasome mediated innate immune response in cholestatic liver disease remain elusive. Here we investigated the functional role of gut-liver crosstalk for cholestatic liver disease in the murine Mdr2 knockout (Mdr2-/-) model resembling human primary sclerosing cholangitis (PSC).

Methods:

Mdr2-/-, Mdr2-/- crossed with hepatocyte-specific deletion of capsase-8 (Mdr2-/-/Casp8Δhepa) and wildtype (WT) control mice were housed for 8w or 52w respectively to characterize the impact of Mdr2 deletion on liver and gut including comprehensive bile acid and microbiota profiling. To block caspase activation, a pan-caspase inhibitor (IDN-7314) was administered. Finally, the functional role of Mdr2-/- associated intestinal dysbiosis was studied by microbiota transfer (FMT) experiments.

Results:

Mdr2-/- mice displayed an unfavorable intestinal microbiota signature and pronounced NLRP3 inflammasome activation within the gut-liver axis, as found by immunostaining and western blot analysis in the intestine as well as in the liver. Intestinal dysbiosis in Mdr2-/- mice prompted intestinal barrier dysfunction evidenced by reduced colonic mucus layers, reduction of tight junction expression and increased permeability evidenced by an in-vivo FITC-dextran assay. Loss of intestinal barrier integrity and bacterial translocation triggered the hepatic NLRP3 mediated innate immune response and fueled liver disease progression. Strinkingly, transfer of Mdr2-/- microbiota into healthy WT control mice, urged intestinal barrier impairment and induced significant liver injury in recipient mice, highlighting the causal role of intestinal dysbiosis for disease progression in Mdr2-/- mice.

This phenotype could not be rescued by introducing Mdr2-/-/Casp8Δhepa indicating that hepatocytic caspase-8 activation is a downstream consequence and dispensable for the inflammatory response. In contrast, caspase inhibition via IDN-7314 dampened inflammasome activation, improved intestinal barrier function, ameliorated liver injury, reversed serum bile acid profile and cholestasis associated microbiota signature.

Conclusion:

Cholestatic liver disease in Mdr2-/- mice triggers intestinal dysbiosis, which is transmissible to healthy WT mice. In turn, translocation of endotoxin into the portal vein and subsequent NLRP3 inflammasome activation contribute to higher liver injury in Mdr2-/- mice. This process does not essentially depend on hepatocytic caspase-8, but can be blocked by IDN-7314, highlighting the central role of the inflammasome mediated innate immune within the gut-liver axis.