Spontaneous episodic inflammation in the intestines of mice lacking HNF4A is driven by microbiota and associated with early life microbiota alterations
Authors: Kelly Cecelia , Jawahar Jayanth , Davey Lauren , Everitt Jeffrey I. , Galanko Joseph A. , Anderson Chelsea , Avendano Jonathan E. , McCann Jessica R. , Sartor R. Balfour , Valdivia Raphael H. , Rawls John F. 
Published: August 2023

ABSTRACT

The inflammatory bowel diseases (IBD) occur in genetically susceptible individuals who mount inappropriate immune responses to their microbiota leading to chronic intestinal inflammation. Whereas IBD clinical presentation is well described, how interactions between microbiota and host genotype impact early subclinical stages of the disease remains unclear. The transcription factor hepatocyte nuclear factor 4 alpha (HNF4A) has been associated with human IBD, and deletion of
Hnf4a
in intestinal epithelial cells (IECs) in mice (
Hnf4a
ΔIEC
) leads to spontaneous colonic inflammation by 6–12 mo of age. Here, we tested if pathology in
Hnf4a
ΔIEC
mice begins earlier in life and if microbiota contribute to that process. Longitudinal analysis revealed that
Hnf4a
ΔIEC
mice reared in specific pathogen-free (SPF) conditions develop episodic elevated fecal lipocalin 2 (Lcn2) and loose stools beginning by 4–5 wk of age. Lifetime cumulative Lcn2 levels correlated with histopathological features of colitis at 12 mo. Antibiotic and gnotobiotic tests showed that these phenotypes in
Hnf4a
ΔIEC
mice were dependent on microbiota. Fecal 16S rRNA gene sequencing in SPF
Hnf4a
ΔIEC
and control mice disclosed that genotype significantly contributed to differences in microbiota composition by 12 mo, and longitudinal analysis of the
Hnf4a
ΔIEC
mice with the highest lifetime cumulative Lcn2 revealed that microbial community differences emerged early in life when elevated fecal Lcn2 was first detected. These microbiota differences included enrichment of a novel phylogroup of
Akkermansia muciniphila
in
Hnf4a
ΔIEC
mice. We conclude that HNF4A functions in IEC to shape composition of the gut microbiota and protect against episodic inflammation induced by microbiota throughout the lifespan.

IMPORTANCE
The inflammatory bowel diseases (IBD), characterized by chronic inflammation of the intestine, affect millions of people around the world. Although significant advances have been made in the clinical management of IBD, the early subclinical stages of IBD are not well defined and are difficult to study in humans. This work explores the subclinical stages of disease in mice lacking the IBD-associated transcription factor HNF4A in the intestinal epithelium. Whereas these mice do not develop overt disease until late in adulthood, we find that they display episodic intestinal inflammation, loose stools, and microbiota changes beginning in very early life stages. Using germ-free and antibiotic-treatment experiments, we reveal that intestinal inflammation in these mice was dependent on the presence of microbiota. These results suggest that interactions between host genotype and microbiota can drive early subclinical pathologies that precede the overt onset of IBD and describe a mouse model to explore those important processes.