title: Chondroitin sulfate proteoglycan 4 functions as the cellular receptor for Clostridium difficile toxin B.
abstract: As a gram-positive, spore-forming anaerobic bacillus, Clostridium difficile (C. difficile) is responsible for severe and fatal pseudomembranous colitis, and poses the most urgent antibiotic resistance threat worldwide. Epidemic C. difficile is the leading cause of antibiotic-associated diarrhoea globally, especially diarrhoea due to the emergence of hypervirulent strains associated with high mortality and morbidity. TcdB, one of the key virulence factors secreted by this bacterium, enters host cells through a poorly understood mechanism to elicit its pathogenic effect. Here we report the first identification of the TcdB cellular receptor, chondroitin sulfate proteoglycan 4 (CSPG4). CSPG4 was initially isolated from a whole-genome human shRNAmir library screening, and its role was confirmed by both TALEN- and CRISPR/Cas9-mediated gene knockout in human cells. CSPG4 is critical for TcdB binding to the cell surface, inducing cytoskeleton disruption and cell death. A direct interaction between the N-terminus of CSPG4 and the C-terminus of TcdB was confirmed, and the soluble peptide of the toxin-binding domain of CSPG4 could protect cells from the action of TcdB. Notably, the complete loss of CSPG4/NG2 decreased TcdB-triggered interleukin-8 induction in mice without significantly affecting animal mortality. Based on both the in vitro and in vivo studies, we propose a dual-receptor model for TcdB endocytosis. The discovery of the first TcdB receptor reveals a previously unsuspected role for CSPG4 and provides a new therapeutic target for the treatment of C. difficile infection.
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Yuan
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Hongmin
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Changzu
authors.initials: C
authors.lastname: Cai
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Shiyou
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Yuexin
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Xiaozhou
authors.initials: X
authors.lastname: Yang
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Ruina
authors.initials: R
authors.lastname: He
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Chan
authors.initials: C
authors.lastname: Li
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Shengjie
authors.initials: S
authors.lastname: Guo
authors.affiliation: School of Bioscience and Biotechnology, South China University of Technology, Guangzhou, Guangdong 510006, China.
authors.forename: Shan
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Microbial Pathogenesis, University of Maryland Dental School, Baltimore, Maryland 21201, USA.
authors.forename: Tuxiong
authors.initials: T
authors.lastname: Huang
authors.affiliation: Department of Microbial Pathogenesis, University of Maryland Dental School, Baltimore, Maryland 21201, USA.
authors.forename: Gregorio
authors.initials: G
authors.lastname: Perez-Cordon
authors.affiliation: Department of Microbial Pathogenesis, University of Maryland Dental School, Baltimore, Maryland 21201, USA.
authors.forename: Hanping
authors.initials: H
authors.lastname: Feng
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Wensheng
authors.initials: W
authors.lastname: Wei
mesh_terms: D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001427:Bacterial Toxins / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D016360:Clostridioides difficile / Q000378:metabolism*; D003015:Clostridium Infections / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D003599:Cytoskeleton / Q000378:metabolism; D004705:Endocytosis; D055786:Gene Knockout Techniques; D057809:HEK293 Cells; D019073:HT29 Cells; D006367:HeLa Cells; D006801:Humans; D016209:Interleukin-8 / Q000097:blood; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011509:Proteoglycans / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D000099216:Chondroitin Sulfate Proteoglycan 4
chemical_list: D000941:Antigens; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D016209:Interleukin-8; D011509:Proteoglycans; D034741:RNA, Small Interfering; D000099216:Chondroitin Sulfate Proteoglycan 4; C031815:toxB protein, Clostridium difficile