Sulfated glycosaminoglycans and low-density lipoprotein receptor contribute to Clostridium difficile toxin A entry into cells

Liang Tao, Songhai Tian, Jie Zhang, Zhuoming Liu, Lindsey Robinson-McCarthy, Shin Ichiro Miyashita, David T. Breault, Ralf Gerhard, Siam Oottamasathien, Sean P.J. Whelan, Min Dong

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Clostridium difficile toxin A (TcdA) is a major exotoxin contributing to disruption of the colonic epithelium during C. difficile infection. TcdA contains a carbohydrate-binding combined repetitive oligopeptides (CROPs) domain that mediates its attachment to cell surfaces, but recent data suggest the existence of CROPs-independent receptors. Here, we carried out genome-wide clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated protein 9 (Cas9)-mediated screens using a truncated TcdA lacking the CROPs, and identified sulfated glycosaminoglycans (sGAGs) and low-density lipoprotein receptor (LDLR) as host factors contributing to binding and entry of TcdA. TcdA recognizes the sulfation group in sGAGs. Blocking sulfation and glycosaminoglycan synthesis reduces TcdA binding and entry into cells. Binding of TcdA to the colonic epithelium can be reduced by surfen, a small molecule that masks sGAGs, by GM-1111, a sulfated heparan sulfate analogue, and by sulfated cyclodextrin, a sulfated small molecule. Cells lacking LDLR also show reduced sensitivity to TcdA, although binding between LDLR and TcdA are not detected, suggesting that LDLR may facilitate endocytosis of TcdA. Finally, GM-1111 reduces TcdA-induced fluid accumulation and tissue damage in the colon in a mouse model in which TcdA is injected into the caecum. These data demonstrate in vivo and pathological relevance of TcdA–sGAGs interactions, and reveal a potential therapeutic approach of protecting colonic tissues by blocking these interactions.

Original languageEnglish
Pages (from-to)1760-1769
Number of pages10
JournalNature microbiology
Volume4
Issue number10
DOIs
StatePublished - Oct 1 2019
Externally publishedYes

Fingerprint Dive into the research topics of 'Sulfated glycosaminoglycans and low-density lipoprotein receptor contribute to Clostridium difficile toxin A entry into cells'. Together they form a unique fingerprint.

  • Cite this

    Tao, L., Tian, S., Zhang, J., Liu, Z., Robinson-McCarthy, L., Miyashita, S. I., Breault, D. T., Gerhard, R., Oottamasathien, S., Whelan, S. P. J., & Dong, M. (2019). Sulfated glycosaminoglycans and low-density lipoprotein receptor contribute to Clostridium difficile toxin A entry into cells. Nature microbiology, 4(10), 1760-1769. https://doi.org/10.1038/s41564-019-0464-z