Bile acids and ceramide overcome the entry restriction for GII.3 human norovirus replication in human intestinal enteroids

Kosuke Murakami, Victoria R. Tenge, Umesh C. Karandikar, Shih Ching Lin, Sasirekha Ramani, Khalil Ettayebi, Sue E. Crawford, Xi Lei Zeng, Frederick H. Neill, B. Vijayalakshmi Ayyar, Kazuhiko Katayama, David Y. Graham, Erhard Bieberich, Robert L. Atmar, Mary K. Estes

Research output: Contribution to journalArticlepeer-review

78 Scopus citations

Abstract

Human noroviruses (HuNoVs) cause sporadic and epidemic outbreaks of gastroenteritis in all age groups worldwide. We previously reported that stem cell-derived human intestinal enteroid (HIE) cultures support replication of multiple HuNoV strains and that some strains (e.g., GII.3) replicate only in the presence of bile. Heatand trypsin-treatment of bile did not reduce GII.3 replication, indicating a nonproteinaceous component in bile functions as an active factor. Here we show that bile acids (BAs) are critical for GII.3 replication and replication correlates with BA hydrophobicity. Using the highly effective BA, glycochenodeoxycholic acid (GCDCA), we show BAs act during the early stage of infection, BA-dependent replication in HIEs is not mediated by detergent effects or classic farnesoid X receptor or Takeda G protein-coupled receptor 5 signaling but involves another G protein-coupled receptor, sphingosine-1- phosphate receptor 2, and BA treatment of HIEs increases particle uptake. We also demonstrate that GCDCA induces multiple cellular responses that promote GII.3 replication in HIEs, including enhancement of 1) endosomal uptake, 2) endosomal acidification and subsequent activity of endosomal/lysosomal enzyme acid sphingomyelinase (ASM), and 3) ceramide levels on the apical membrane. Inhibitors of endosomal acidification or ASM reduce GII.3 infection and exogenous addition of ceramide alone permits infection. Furthermore, inhibition of lysosomal exocytosis of ASM, which is required for ceramide production at the apical surface, decreases GII.3 infection. Together, our results support a model where GII.3 exploits rapid BA-mediated cellular endolysosomal dynamic changes and cellular ceramide to enter and replicate in jejunal HIEs.

Original languageEnglish
Pages (from-to)1700-1710
Number of pages11
JournalProceedings of the National Academy of Sciences of the United States of America
Volume117
Issue number3
DOIs
StatePublished - Jan 21 2020

Bibliographical note

Publisher Copyright:
© 2020 National Academy of Sciences. All rights reserved.

Keywords

  • Acid sphingomyelinase
  • Bile acid
  • Ceramide
  • Enteroid/organoid
  • Norovirus

ASJC Scopus subject areas

  • General

Fingerprint

Dive into the research topics of 'Bile acids and ceramide overcome the entry restriction for GII.3 human norovirus replication in human intestinal enteroids'. Together they form a unique fingerprint.

Cite this