Zranb1-mutant mice display abnormal colonic mucus production and exacerbation of DSS-induced colitis

Akiko Tamura; Go Ito; Hiroki Matsuda; Yoichi Nibe-Shirakihara; Yuichi Hiraoka; Sayuki Kitagawa; Yui Hiraguri; Sayaka Nagata; Emi Aonuma; Kana Otsubo; Yasuhiro Nemoto; Takashi Nagaishi; Mamoru Watanabe; Ryuichi Okamoto; Shigeru Oshima

doi:10.1016/j.bbrc.2022.08.046

Zranb1-mutant mice display abnormal colonic mucus production and exacerbation of DSS-induced colitis

Biochem Biophys Res Commun. 2022 Nov 5:628:147-154. doi: 10.1016/j.bbrc.2022.08.046. Epub 2022 Aug 28.

Authors

Affiliations

¹ Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
² Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan; Advanced Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
³ Department of Pathological Cell Biology, Medical Research Institute, Tokyo Medical and Dental University, Japan.
⁴ Department of Molecular Neuroscience, Medical Research Institute, Tokyo Medical and Dental University, Japan; Laboratory of Genome Editing for Biomedical Research, Medical Research Institute, Tokyo Medical and Dental University, Japan.
⁵ Department of Advanced Therapeutics for G.I. Diseases, Tokyo Medical and Dental University, Tokyo, Japan.
⁶ Advanced Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
⁷ Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan; Institute of Research, Tokyo Medical and Dental University, Tokyo, Japan. Electronic address: soshima.gast@tmd.ac.jp.

PMID: 36087511
DOI: 10.1016/j.bbrc.2022.08.046

Abstract

Expression of mucin MUC2, a component of the colonic mucus layer, plays a crucial role in intestinal homeostasis. Here, we describe a new regulator of MUC2 expression, the deubiquitinase ZRANB1 (Trabid). A ZRANB1 mutation changing cysteine to serine in amino acid position 443, affects ubiquitination. To analyze ZRANB1 function in the intestine, we generated Zranb1 C443S mutant knock-in (Zranb1^C443S/C443S) mice using the CRISPR/Cas9 system. Zranb1^C443S/C443S mice exhibited decreased mRNA expression and MUC2 production. Colonic organoids from Zranb1^C443S/C443S mice displayed decreased Muc2 mRNA expression following differentiation into goblet cells. Finally, we analyzed dextran sulfate sodium-induced colitis to understand ZRANB1's role in intestinal inflammation. Zranb1^C443S/C443S mice with colitis exhibited significant weight loss, reduced colon length, and worsening clinical and pathological scores, indicating that ZRANB1 contributes to intestinal homeostasis. Together, these results suggest that ZRANB1 regulates MUC2 expression and intestinal inflammation, which may help elucidating the pathogenesis of inflammatory bowel disease and developing new therapeutics targeting ZRANB1.

Keywords: Colitis; IBD; Muc2; Organoid; Ubiquitin; ZRANB1.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Colitis* / chemically induced
Colitis* / genetics
Colitis* / metabolism
Cysteine / metabolism
Deubiquitinating Enzymes / metabolism
Dextran Sulfate / toxicity
Inflammation / pathology
Intestinal Mucosa* / metabolism
Intestinal Mucosa* / pathology
Mice
Mucins / metabolism
Mucus / metabolism
RNA, Messenger / genetics
Serine / metabolism
Ubiquitin-Specific Proteases* / genetics
Ubiquitin-Specific Proteases* / metabolism

Substances

Mucins
RNA, Messenger
Serine
Dextran Sulfate
Deubiquitinating Enzymes
Ubiquitin-Specific Proteases
Zranb1 protein, mouse
Cysteine