Abstract
Objective Conflicting microbiota data exist for primary sclerosing cholangitis (PSC) and experimental models. Goal: Define complex interactions between resident microbes and their association in PSC patients by studying antibiotic-treated specific pathogen-free (SPF) and germ-free (GF) multi-drug-resistant 2 deficient (mdr2-/-) mice.
Design We measured weights, liver enzymes, RNA expression, histological, immunohistochemical and fibrotic biochemical parameters, fecal 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 exaggerated hepatic inflammation and fibrosis with 100% mortality by 8 weeks; early SPF autologous stool transplantation rescued liver-related mortality. Broad-spectrum antibiotics and vancomycin alone accelerated disease in weanling SPF mdr2-/- mice, indicating that vancomycin-sensitive resident microbiota protect against hepatobiliary disease. Vancomycin treatment selectively decreased Lachnospiraceae and short-chain fatty acids (SCFAs) but expanded Enterococcus and Enterobacteriaceae. Antibiotics increased cytolysin-expressing E. faecalis and E. coli liver translocation; colonization of gnotobiotic mdr2-/- mice with translocated E. faecalis and E. coli strains accelerated liver inflammation and mortality. Lachnospiraceae colonization of antibiotic pre-treated mdr2-/- mice reduced liver fibrosis, inflammation and translocation of pathobionts, while Lachnospiraceae-produced SCFA decreased fibrosis. Fecal E. faecalis/ Enterobacteriaceae was positively and Lachnospiraceae was negatively associated with PSC patients’ clinical severity Mayo risk scores.
Conclusions We identified specific functionally protective and detrimental resident bacterial species in mdr2-/- mice and PSC patients with associated clinical outcomes. These insights may guide personalized targeted therapeutic interventions in PSC patients.
Competing Interest Statement
The authors have declared no competing interest.
Abbreviations
- Abx
- broad-spectrum antibiotics
- ALP
- alkaline phosphatase
- ALT
- alanine aminotransferase
- BA
- bile acids
- BSH
- bile salt hydrolase
- CA
- Cholic Acid
- CDCA
- chenodeoxycholic acid
- col1a1
- collagen 1a1
- cyto
- cytolysin
- DCA
- Deoxycholic Acid
- FMT
- fecal microbiota transplant
- GF
- germ-free
- HYP
- hepatic hydroxyproline
- Lachno
- Lachnospiraceae
- LC
- Lithocholic Acid
- Lcn-2
- lipocalin-2
- M
- metronidazole
- MCP
- monocyte chemoattractant protein
- mdr2-/-
- multi-drug resistant 2 gene knockout
- MCA
- muricholic acid
- N
- neomycin
- OTU
- operational taxonomic unit
- PSC
- Primary Sclerosing Cholangitis
- SCFA
- short-chain fatty acids
- SPF
- specific pathogen-free
- TIMP-1
- tissue inhibitor of metalloproteinase-1
- TB
- Total bilirubin
- V
- vancomycin
- WT
- wild-type