Bile acids promote the caveolae-associated entry of swine acute diarrhea syndrome coronavirus in porcine intestinal enteroids

Author summaryBile acids (BAs), a commonly studied category of microbial metabolites, have long been acknowledged to have proviral or antiviral activities. Recent studies using different swine enteric coronaviruses (SECoVs) showed that BA play an important role in regulating viral replication in vitro. A mechanistic understanding of how BA regulates SECoV replication in small intestinal enterocytes is lacking. Herein, we utilized an emerging highly pathogenic SECoV, swine acute diarrhea syndrome (SADS)-CoV, which possesses the potential for zoonotic transmission, to investigate the crucial role of BA in modulating viral replication in porcine intestinal enteroids (PIEs). Our observations explain how BAs acts on epithelial cells to enhance SADS-CoV replication by inducing caveolae-mediated endocytosis and endosomal acidification, altering the dynamics of viral trafficking through the cellular endosomal/lysosomal system. Our results shed light on the role of BAs in the rapid establishment of SECoV infection within the intestinal microenvironment. Intestinal microbial metabolites have been increasingly recognized as important regulators of enteric viral infection. However, very little information is available about which specific microbiota-derived metabolites are crucial for swine enteric coronavirus (SECoV) infection in vivo. Using swine acute diarrhea syndrome (SADS)-CoV as a model, we were able to identify a greatly altered bile acid (BA) profile in the small intestine of infected piglets by untargeted metabolomic analysis. Using a newly established ex vivo model-the stem cell-derived porcine intestinal enteroid (PIE) culture-we demonstrated that certain BAs, cholic acid (CA) in particular, enhance SADS-CoV replication by acting on PIEs at the early phase of infection. We ruled out the possibility that CA exerts an augmenting effect on viral replication through classic farnesoid X receptor or Takeda G protein-coupled receptor 5 signaling, innate immune suppression or viral attachment. BA induced multiple cellular responses including rapid changes in caveolae-mediated endocytosis, endosomal acidification and dynamics of the endosomal/lysosomal system that are critical for SADS-CoV replication. Thus, our findings shed light on how SECoVs exploit microbiome-derived metabolite BAs to swiftly establish viral infection and accelerate replication within the intestinal microenvironment.