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Published in: Digestive Diseases and Sciences 12/2022

21-03-2022 | Original Article

UBE2W Improves the Experimental Colitis by Inhibiting the NF-κB Signaling Pathway

Authors: Shaoxin Wang, Jiang Pu, Xiaowei Li, Zhihui Yan, Chao Li, Yan Zheng, Zhe Luo, Lihong Cui

Published in: Digestive Diseases and Sciences | Issue 12/2022

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Abstract

Background

The NF-κB signaling cascade regulates immune response and is often dysregulated in tumor development. UBE2W is a novel type I ubiquitin-conjugating enzyme (E2) whose biological function is still unclear.

Aims

This study was designed to investigate whether UBE2W regulates NF-κB signaling pathway and is involved in the progression of experimental colitis.

Methods

At the cellular level, the effect of UBE2W on NF-κB transcriptional activity was measured using a dual-luciferase reporter assay. The influence of UBE2W on NF-κB pathway activation and the entry of p65 into the nucleus were determined by Western blot and immunofluorescence analyses, respectively. Moreover, the colitis model was established by administering 2.5% dextran sulfate sodium (DSS)/water to UBE2W overexpression, UBE2W-knockdown and control mice. Body weight, stool consistency, colon length and clinical severity were examined. Expression of pro-inflammatory cytokines and phosphorylation of p65 and IκB in the colon tissue were measured by qRT-PCR and Western blot, respectively.

Results

UBE2W inhibited TNFα-induced NF-κB transcription activity, attenuated IκB and p65 phosphorylation, downregulated TNFα and IL-8 expression and blocked the entry of p65 into the nucleus. In the DSS-induced colitis model, UBE2W-knockdown mice had increased weight loss, more serious diarrhea and mucosal injures compared with the control mice. Moreover, phosphorylation of IκB and p65 and the expression of pro-inflammatory mediators such as TNFα, IL-6 were significantly increased in UBE2W knockdown mice. However, these changes were completely reversed in UBE2W overexpression mice.

Conclusions

The overexpression of UBE2W ameliorates the severity of DSS-induced colitis, which may be mediated by inhibiting the expression of pro-inflammatory mediators and activation of the NF-κB signaling pathway. These findings provide evidence that UBE2W might have potential therapeutic implications in IBD.
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Literature
2.
go back to reference Tilborghs S, Corthouts J, Verhoeven Y et al. The role of nuclear factor-kappa B signaling in human cervical cancer. Crit Rev Oncol Hematol. 2017;120:141–150.CrossRefPubMed Tilborghs S, Corthouts J, Verhoeven Y et al. The role of nuclear factor-kappa B signaling in human cervical cancer. Crit Rev Oncol Hematol. 2017;120:141–150.CrossRefPubMed
3.
go back to reference Ghosh S, Dass JFP. Study of pathway cross-talk interactions with NF-kappaB leading to its activation via ubiquitination or phosphorylation: a brief review. Gene. 2016;584:97–109.CrossRefPubMed Ghosh S, Dass JFP. Study of pathway cross-talk interactions with NF-kappaB leading to its activation via ubiquitination or phosphorylation: a brief review. Gene. 2016;584:97–109.CrossRefPubMed
4.
go back to reference Aksentijevich I, Zhou Q. NF-kappaB pathway in autoinflammatory diseases: dysregulation of protein modifications by ubiquitin defines a new category of autoinflammatory diseases. Front Immunol. 2017;8:399.CrossRefPubMedPubMedCentral Aksentijevich I, Zhou Q. NF-kappaB pathway in autoinflammatory diseases: dysregulation of protein modifications by ubiquitin defines a new category of autoinflammatory diseases. Front Immunol. 2017;8:399.CrossRefPubMedPubMedCentral
5.
go back to reference Christensen DE, Brzovic PS, Klevit RE. E2-BRCA1 RING interactions dictate synthesis of mono- or specific polyubiquitin chain linkages. Nat Struct Mol Biol. 2007;14:941–948.CrossRefPubMed Christensen DE, Brzovic PS, Klevit RE. E2-BRCA1 RING interactions dictate synthesis of mono- or specific polyubiquitin chain linkages. Nat Struct Mol Biol. 2007;14:941–948.CrossRefPubMed
6.
go back to reference Zeng L, Tan J, Xue M et al. An engineering probiotic producing defensin-5 ameliorating dextran sodium sulfate-induced mice colitis via Inhibiting NF-kB pathway. J Transl Med. 2020;18:107.CrossRefPubMedPubMedCentral Zeng L, Tan J, Xue M et al. An engineering probiotic producing defensin-5 ameliorating dextran sodium sulfate-induced mice colitis via Inhibiting NF-kB pathway. J Transl Med. 2020;18:107.CrossRefPubMedPubMedCentral
7.
go back to reference Kim SW, Kim HM, Yang KM et al. Bifidobacterium lactis inhibits NF-kappaB in intestinal epithelial cells and prevents acute colitis and colitis-associated colon cancer in mice. Inflamm Bowel Dis. 2010;16:1514–1525.CrossRefPubMed Kim SW, Kim HM, Yang KM et al. Bifidobacterium lactis inhibits NF-kappaB in intestinal epithelial cells and prevents acute colitis and colitis-associated colon cancer in mice. Inflamm Bowel Dis. 2010;16:1514–1525.CrossRefPubMed
8.
go back to reference Wirtz S, Neufert C, Weigmann B et al. Chemically induced mouse models of intestinal inflammation. Nat Protoc. 2007;2:541–546.CrossRefPubMed Wirtz S, Neufert C, Weigmann B et al. Chemically induced mouse models of intestinal inflammation. Nat Protoc. 2007;2:541–546.CrossRefPubMed
10.
go back to reference Farooq SM, Stillie R, Svensson M et al. Therapeutic effect of blocking CXCR2 on neutrophil recruitment and dextran sodium sulfate-induced colitis. J Pharmacol Exp Ther. 2009;329:123–129.CrossRefPubMed Farooq SM, Stillie R, Svensson M et al. Therapeutic effect of blocking CXCR2 on neutrophil recruitment and dextran sodium sulfate-induced colitis. J Pharmacol Exp Ther. 2009;329:123–129.CrossRefPubMed
11.
12.
go back to reference Maeda S, Hsu LC, Liu H et al. Nod2 mutation in Crohn’s disease potentiates NF-kappaB activity and IL-1beta processing. Science. 2005;307:734–738.CrossRefPubMed Maeda S, Hsu LC, Liu H et al. Nod2 mutation in Crohn’s disease potentiates NF-kappaB activity and IL-1beta processing. Science. 2005;307:734–738.CrossRefPubMed
13.
go back to reference Tan B, Mu R, Chang Y et al. RNF4 negatively regulates NF-kappaB signaling by down-regulating TAB2. FEBS Lett. 2015;589:2850–2858.CrossRefPubMed Tan B, Mu R, Chang Y et al. RNF4 negatively regulates NF-kappaB signaling by down-regulating TAB2. FEBS Lett. 2015;589:2850–2858.CrossRefPubMed
14.
go back to reference Pan JX. LncRNA H19 promotes atherosclerosis by regulating MAPK and NF-kB signaling pathway. Eur Rev Med Pharmacol Sci. 2017;21:322–328.PubMed Pan JX. LncRNA H19 promotes atherosclerosis by regulating MAPK and NF-kB signaling pathway. Eur Rev Med Pharmacol Sci. 2017;21:322–328.PubMed
16.
go back to reference Wang S, Lin Y, Yuan X et al. REV-ERBalpha integrates colon clock with experimental colitis through regulation of NF-kappaB/NLRP3 axis. Nat Commun. 2018;9:4246.CrossRefPubMedPubMedCentral Wang S, Lin Y, Yuan X et al. REV-ERBalpha integrates colon clock with experimental colitis through regulation of NF-kappaB/NLRP3 axis. Nat Commun. 2018;9:4246.CrossRefPubMedPubMedCentral
17.
go back to reference Edelblum KL, Washington MK, Koyama T et al. Raf protects against colitis by promoting mouse colon epithelial cell survival through NF-kappaB. Gastroenterology. 2008;135:539–551.CrossRefPubMed Edelblum KL, Washington MK, Koyama T et al. Raf protects against colitis by promoting mouse colon epithelial cell survival through NF-kappaB. Gastroenterology. 2008;135:539–551.CrossRefPubMed
18.
go back to reference Lee CY, Lai TY, Tsai MK et al. The ubiquitin ligase ZNRF1 promotes caveolin-1 ubiquitination and degradation to modulate inflammation. Nat Commun. 2017;8:15502.CrossRefPubMedPubMedCentral Lee CY, Lai TY, Tsai MK et al. The ubiquitin ligase ZNRF1 promotes caveolin-1 ubiquitination and degradation to modulate inflammation. Nat Commun. 2017;8:15502.CrossRefPubMedPubMedCentral
19.
go back to reference Zhang Y, Zhu H, Zhao L et al. [Generation of mouse UBE2W antibody and analysis of UBE2W expression in mouse tissues. Sheng Wu Gong Cheng Xue Bao. 2008;24:547–552.PubMed Zhang Y, Zhu H, Zhao L et al. [Generation of mouse UBE2W antibody and analysis of UBE2W expression in mouse tissues. Sheng Wu Gong Cheng Xue Bao. 2008;24:547–552.PubMed
20.
go back to reference Yin G, Ji C, Wu T et al. Cloning, characterization and subcellular localization of a gene encoding a human ubiquitin-conjugating enzyme (E2) homologous to the Arabidopsis thaliana UBC-16 gene product. Front Biosci. 2006;11:1500–1507.CrossRefPubMed Yin G, Ji C, Wu T et al. Cloning, characterization and subcellular localization of a gene encoding a human ubiquitin-conjugating enzyme (E2) homologous to the Arabidopsis thaliana UBC-16 gene product. Front Biosci. 2006;11:1500–1507.CrossRefPubMed
21.
go back to reference Soss SE, Rose KL, Hill S et al. Biochemical and proteomic analysis of ubiquitination of Hsc70 and Hsp70 by the E3 Ligase CHIP. PLoS One. 2015;10:e0128240.CrossRefPubMedPubMedCentral Soss SE, Rose KL, Hill S et al. Biochemical and proteomic analysis of ubiquitination of Hsc70 and Hsp70 by the E3 Ligase CHIP. PLoS One. 2015;10:e0128240.CrossRefPubMedPubMedCentral
23.
go back to reference Zhang Y, Zhou X, Zhao L et al. UBE2W interacts with FANCL and regulates the monoubiquitination of Fanconi anemia protein FANCD2. Mol Cells. 2011;31:113–122.CrossRefPubMedPubMedCentral Zhang Y, Zhou X, Zhao L et al. UBE2W interacts with FANCL and regulates the monoubiquitination of Fanconi anemia protein FANCD2. Mol Cells. 2011;31:113–122.CrossRefPubMedPubMedCentral
24.
go back to reference Vittal V, Wenzel DM, Brzovic PS et al. Biochemical and structural characterization of the ubiquitin-conjugating enzyme UBE2W reveals the formation of a noncovalent homodimer. Cell Biochem Biophys. 2013;67:103–110.CrossRefPubMedPubMedCentral Vittal V, Wenzel DM, Brzovic PS et al. Biochemical and structural characterization of the ubiquitin-conjugating enzyme UBE2W reveals the formation of a noncovalent homodimer. Cell Biochem Biophys. 2013;67:103–110.CrossRefPubMedPubMedCentral
25.
go back to reference Tatham MH, Plechanovova A, Jaffray EG et al. Ube2W conjugates ubiquitin to alpha-amino groups of protein N-termini. Biochem J. 2013;453:137–145.CrossRefPubMed Tatham MH, Plechanovova A, Jaffray EG et al. Ube2W conjugates ubiquitin to alpha-amino groups of protein N-termini. Biochem J. 2013;453:137–145.CrossRefPubMed
26.
go back to reference Hoffmann M, Schwertassek U, Seydel A et al. A refined and translationally relevant model of chronic DSS colitis in BALB/c mice. Lab Anim. 2018;52:240–252.CrossRefPubMed Hoffmann M, Schwertassek U, Seydel A et al. A refined and translationally relevant model of chronic DSS colitis in BALB/c mice. Lab Anim. 2018;52:240–252.CrossRefPubMed
27.
go back to reference Gong Z, Zhao S, Zhou J et al. Curcumin alleviates DSS-induced colitis via inhibiting NLRP3 inflammsome activation and IL-1beta production. Mol Immunol. 2018;104:11–19.CrossRefPubMed Gong Z, Zhao S, Zhou J et al. Curcumin alleviates DSS-induced colitis via inhibiting NLRP3 inflammsome activation and IL-1beta production. Mol Immunol. 2018;104:11–19.CrossRefPubMed
28.
go back to reference Pichai MV, Ferguson LR. Potential prospects of nanomedicine for targeted therapeutics in inflammatory bowel diseases. World J Gastroenterol. 2012;18:2895–2901.CrossRefPubMedPubMedCentral Pichai MV, Ferguson LR. Potential prospects of nanomedicine for targeted therapeutics in inflammatory bowel diseases. World J Gastroenterol. 2012;18:2895–2901.CrossRefPubMedPubMedCentral
29.
go back to reference Ramakrishnan SK, Zhang H, Ma X et al. Intestinal non-canonical NFkappaB signaling shapes the local and systemic immune response. Nat Commun. 2019;10:660.CrossRefPubMedPubMedCentral Ramakrishnan SK, Zhang H, Ma X et al. Intestinal non-canonical NFkappaB signaling shapes the local and systemic immune response. Nat Commun. 2019;10:660.CrossRefPubMedPubMedCentral
30.
go back to reference Shi Y, Guo Y, Zhou J et al. Herbs-partitioned moxibustion improves intestinal epithelial tight junctions by upregulating A20 expression in a mouse model of Crohn’s disease. Biomed Pharmacother. 2019;118:109149.CrossRefPubMed Shi Y, Guo Y, Zhou J et al. Herbs-partitioned moxibustion improves intestinal epithelial tight junctions by upregulating A20 expression in a mouse model of Crohn’s disease. Biomed Pharmacother. 2019;118:109149.CrossRefPubMed
31.
go back to reference Chen W, Chen W, Zhu J, Chen N, Lu Y. Potent anti-inflammatory activity of tetramethylpyrazine is mediated through suppression of NF-k. Iran J Pharm Res. 2016;15:197–204.PubMedPubMedCentral Chen W, Chen W, Zhu J, Chen N, Lu Y. Potent anti-inflammatory activity of tetramethylpyrazine is mediated through suppression of NF-k. Iran J Pharm Res. 2016;15:197–204.PubMedPubMedCentral
32.
go back to reference Kondylis V, Kumari S, Vlantis K, Pasparakis M. The interplay of IKK, NF-kappaB and RIPK1 signaling in the regulation of cell death, tissue homeostasis and inflammation. Immunol Rev. 2017;277:113–127.CrossRefPubMed Kondylis V, Kumari S, Vlantis K, Pasparakis M. The interplay of IKK, NF-kappaB and RIPK1 signaling in the regulation of cell death, tissue homeostasis and inflammation. Immunol Rev. 2017;277:113–127.CrossRefPubMed
33.
go back to reference Lei B, Xie L, Zhang S et al. UBE2W down-regulation promotes cell apoptosis and correlates with hypospermatogenesis. Andrologia. 2020;52:e13474.CrossRefPubMed Lei B, Xie L, Zhang S et al. UBE2W down-regulation promotes cell apoptosis and correlates with hypospermatogenesis. Andrologia. 2020;52:e13474.CrossRefPubMed
Metadata
Title
UBE2W Improves the Experimental Colitis by Inhibiting the NF-κB Signaling Pathway
Authors
Shaoxin Wang
Jiang Pu
Xiaowei Li
Zhihui Yan
Chao Li
Yan Zheng
Zhe Luo
Lihong Cui
Publication date
21-03-2022
Publisher
Springer US
Published in
Digestive Diseases and Sciences / Issue 12/2022
Print ISSN: 0163-2116
Electronic ISSN: 1573-2568
DOI
https://doi.org/10.1007/s10620-022-07453-4

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