Protective and aggressive bacterial subsets and metabolites modify hepatobiliary inflammation and fibrosis in a murine model of PSC

Muyiwa Awoniyi; Jeremy Wang; Billy Ngo; Vik Meadows; Jason Tam; Amba Viswanathan; Yunjia Lai; Stephanie Montgomery; Morgan Farmer; Martin Kummen; Louise Thingholm; Christoph Schramm; Corinna Bang; Andre Franke; Kun Lu; Huiping Zhou; Jasmohan S Bajaj; Phillip B Hylemon; Jenny Ting; Yury V Popov; Johannes Roksund Hov; Heather L Francis; Ryan Balfour Sartor

doi:10.1136/gutjnl-2021-326500

Protective and aggressive bacterial subsets and metabolites modify hepatobiliary inflammation and fibrosis in a murine model of PSC

Gut. 2023 Apr;72(4):671-685. doi: 10.1136/gutjnl-2021-326500. Epub 2022 Jun 15.

Authors

Muyiwa Awoniyi^{1

2}, Jeremy Wang^{2

3}, Billy Ngo², Vik Meadows⁴, Jason Tam⁵, Amba Viswanathan², Yunjia Lai⁶, Stephanie Montgomery⁷, Morgan Farmer², Martin Kummen^{8

9}, Louise Thingholm¹⁰, Christoph Schramm¹¹, Corinna Bang¹², Andre Franke¹², Kun Lu^{6

13}, Huiping Zhou^{14

15

16}, Jasmohan S Bajaj^{14

15

16}, Phillip B Hylemon^{14

15

16}, Jenny Ting^{5

17}, Yury V Popov¹⁸, Johannes Roksund Hov^{9

19}, Heather L Francis^{20

21}, Ryan Balfour Sartor^{22

2

5}

Affiliations

¹ Division of Gastroenterology and Hepatology, University of North Carolina System, Chapel Hill, North Carolina, USA.
² Center for Gastrointestinal Biology and Disease, University of North Carolina System, Chapel Hill, North Carolina, USA.
³ Department of Genetics, University of North Carolina System, Chapel Hill, North Carolina, USA.
⁴ Department of Gastroenterology, Indiana University School of Medicine, Indianapolis, Indiana, USA.
⁵ Department of Microbiology and Immunology, University of North Carolina System, Chapel Hill, North Carolina, USA.
⁶ Department of Environmental Sciences and Engineering, Gillings School of Global School of Public Health, University of North Carolina System, Chapel Hill, North Carolina, USA.
⁷ Department of Pathology, Division of Comparative Medicine, and Lineberger Comprehensive Cancer Center, University of North Carolina System, Chapel Hill, North Carolina, USA.
⁸ Norwegian PSC Research Center, Oslo University Hospital, Rikshospitalet, Oslo, Norway.
⁹ Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
¹⁰ Institute of Clinical Molecular Biology, Zentrums für Molekulare Biowissenschaften, Kiel, Schleswig-Holstein, Germany.
¹¹ University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.
¹² Institute of Clinical Molecular Biology, Christian-Albrechts-University of Kiel, Kiel, Germany.
¹³ Department of Environmental Sciences and Engineering, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
¹⁴ Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.
¹⁵ Department of Research, McGuire Veterans Affairs Medical Cente, Richmond, Virginia, USA.
¹⁶ Virginia Commonwealth University Medical Center, Richmond, Virginia, USA.
¹⁷ UNC Lineberger Comprehensive Cancer Center, Center for Translational Immunology, University of North Carolina, Chapel Hill, Chapel Hill, North Carolina, USA.
¹⁸ Department of Gastroenterology, Beth Israel Deaconess Medical Center/Harvard Medical School, Boston, Massachusetts, USA.
¹⁹ Norwegian PSC Research Center, Department of Transplantation Medicine, Oslo University Hospital, Oslo, Norway.
²⁰ Indiana University School of Medicine, Indianapolis, Indiana, USA.
²¹ Richard L. Roudebush VA Medical Center, Indianapolis, Indiana, USA.
²² Division of Gastroenterology and Hepatology, University of North Carolina System, Chapel Hill, North Carolina, USA ryan_balfour_sartor@med.unc.edu.

Abstract

Objective: Conflicting microbiota data exist for primary sclerosing cholangitis (PSC) and experimental models.

Goal: define the function of complex resident microbes and their association relevant to PSC patients by studying germ-free (GF) and antibiotic-treated specific pathogen-free (SPF) multidrug-resistant 2 deficient (mdr2^-/- ) mice and microbial profiles in PSC patient cohorts.

Design: We measured weights, liver enzymes, RNA expression, histological, immunohistochemical and fibrotic biochemical parameters, faecal 16S rRNA gene profiling and metabolomic endpoints in gnotobiotic and antibiotic-treated SPF mdr2^-/- mice and targeted metagenomic analysis in PSC patients.

Results: GF mdr2^-/- mice had 100% mortality by 8 weeks with increasing hepatic bile acid (BA) accumulation and cholestasis. Early SPF autologous stool transplantation rescued liver-related mortality. Inhibition of ileal BA transport attenuated antibiotic-accelerated liver disease and decreased total serum and hepatic BAs. Depletion of vancomycin-sensitive microbiota exaggerated hepatobiliary disease. Vancomycin selectively decreased Lachnospiraceae and short-chain fatty acids (SCFAs) but expanded Enterococcus and Enterobacteriaceae. Antibiotics increased Enterococcus faecalis and Escherichia coli liver translocation. Colonisation of GF mdr2^-/- mice with translocated E. faecalis and E. coli strains accelerated hepatobiliary inflammation and mortality. Lachnospiraceae colonisation of antibiotic pretreated mdr2^-/- mice reduced liver fibrosis, inflammation and translocation of pathobionts, and SCFA-producing Lachnospiraceae and purified SCFA decreased fibrosis. Faecal Lachnospiraceae negatively associated, and E. faecalis/ Enterobacteriaceae positively associated, with PSC patients' clinical severity by Mayo risk scores.

Conclusions: We identified novel functionally protective and detrimental resident bacterial species in mdr2^-/- mice and PSC patients with associated clinical risk score. These insights may guide personalised targeted therapeutic interventions in PSC patients.

Keywords: antibiotics; cholestatic liver diseases; intestinal microbiology; primary sclerosing cholangitis; short chain fatty acids.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Anti-Bacterial Agents / pharmacology
Anti-Bacterial Agents / therapeutic use
Clostridiales
Disease Models, Animal
Escherichia coli*
Inflammation
Liver Cirrhosis
Mice
RNA, Ribosomal, 16S / genetics
Vancomycin*

Substances

Vancomycin
RNA, Ribosomal, 16S
Anti-Bacterial Agents

Abstract

Publication types

MeSH terms

Substances

Grants and funding