Frizzled proteins are colonic epithelial receptors for C. difficile toxin B.
Clostridium difficile toxin B (TcdB) is a critical virulence factor that causes diseases associated with C. difficile infection. Here we carried out CRISPR-Cas9-mediated genome-wide screens and identified the members of the Wnt receptor frizzled family (FZDs) as TcdB receptors. TcdB binds to the conserved Wnt-binding site known as the cysteine-rich domain (CRD), with the highest affinity towards FZD1, 2 and 7. TcdB competes with Wnt for binding to FZDs, and its binding blocks Wnt signalling. FZD1/2/7 triple-knockout cells are highly resistant to TcdB, and recombinant FZD2-CRD prevented TcdB binding to the colonic epithelium. Colonic organoids cultured from FZD7-knockout mice, combined with knockdown of FZD1 and 2, showed increased resistance to TcdB. The colonic epithelium in FZD7-knockout mice was less susceptible to TcdB-induced tissue damage in vivo. These findings establish FZDs as physiologically relevant receptors for TcdB in the colonic epithelium.
Department of Urology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.|Department of Microbiology and Immunobiology and Department of Surgery, Harvard Medical School, Boston, Massachusetts 02115, USA.
Liang
L
Tao
Department of Urology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.|Department of Microbiology and Immunobiology and Department of Surgery, Harvard Medical School, Boston, Massachusetts 02115, USA.
Jie
J
Zhang
Department of Microbiology and Physiological Systems (MaPS), University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
Paul
P
Meraner
Division of Endocrinology, Boston Children's Hospital, Boston, Massachusetts 02115, USA.
Alessio
A
Tovaglieri
Center for Infectious and Inflammatory Diseases, Texas A &M Health Science Center, Houston, Texas 77030, USA.
Xiaoqian
X
Wu
Institute of Toxicology, Hannover Medical School, 30625 Hannover, Germany.
Ralf
R
Gerhard
The F. M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.|Department of Neurology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Xinjun
X
Zhang
Tumor Microenvironment and Cancer Immunology Program, Sanford-Burnham Prebys Medical Discovery Institute, Cancer Center, La Jolla, California 92037, USA.
William B
WB
Stallcup
Division of Endocrinology, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Ji
J
Miao
The F. M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.|Department of Neurology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Xi
X
He
Center for Infectious and Inflammatory Diseases, Texas A &M Health Science Center, Houston, Texas 77030, USA.
Julian G
JG
Hurdle
Division of Endocrinology, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts 02138, USA.
David T
DT
Breault
Department of Microbiology and Physiological Systems (MaPS), University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.|Gastroenterology Division, Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
Abraham L
AL
Brass
Department of Urology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.|Department of Microbiology and Immunobiology and Department of Surgery, Harvard Medical School, Boston, Massachusetts 02115, USA.
Min
M
Dong
D000818:Animals; D000941:Antigens / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001427:Bacterial Toxins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D016466:CHO Cells; D064113:CRISPR-Cas Systems; D002460:Cell Line; D016360:Clostridioides difficile / Q000472:pathogenicity; D003106:Colon / Q000378:metabolism*; D003412:Cricetulus; D004848:Epithelium / Q000378:metabolism*; D005260:Female; D051157:Frizzled Receptors / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D055786:Gene Knockout Techniques; D006801:Humans; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D009940:Organoids / Q000378:metabolism; D000072417:Protein Domains; D011509:Proteoglycans / Q000378:metabolism; D037521:Virulence Factors / Q000378:metabolism; D051153:Wnt Proteins / Q000378:metabolism; D000099216:Chondroitin Sulfate Proteoglycan 4
D000941:Antigens; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D051157:Frizzled Receptors; D009842:Oligopeptides; D011509:Proteoglycans; D037521:Virulence Factors; D051153:Wnt Proteins; D000099216:Chondroitin Sulfate Proteoglycan 4; C031815:toxB protein, Clostridium difficile
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