Epithelial Cell-Intrinsic Notch Signaling Plays an Essential Role in the Maintenance of Gut Immune Homeostasis

Obata, Yuichi; Takahashi, Daisuke; Ebisawa, Masashi; Kakiguchi, Kisa; Yonemura, Shigenobu; Jinnohara, Toshi; Kanaya, Takashi; Fujimura, Y; Ohmae, Masumi; Hase, Koji; Ohno, Hiroshi

doi:10.4049/jimmunol.1101128

Cited by 63 publications

(51 citation statements)

References 39 publications

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“…1) and may in fact play a role in the pathology of Kaiso-induced chronic inflammation at later ages. Similar findings were observed by Obata et al , who demonstrated that Notch-signaling inhibition via IEC-specific deletion of Rbpj (RBP-J IEC/Δ ) resulted in spontaneous inflammation in the colons of these mice [52]. Similar to Kaiso Tg , RBP-J IEC/Δ mice exhibited increased numbers of secretory cells throughout the intestine, as well as augmented neutrophil infiltration in inflamed regions [52].…”

Section: Discussionsupporting

confidence: 79%

Kaiso differentially regulates components of the Notch signaling pathway in intestinal cells

et al. 2017

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BackgroundIn mammalian intestines, Notch signaling plays a critical role in mediating cell fate decisions; it promotes the absorptive (or enterocyte) cell fate, while concomitantly inhibiting the secretory cell fate (i.e. goblet, Paneth and enteroendocrine cells). We recently reported that intestinal-specific Kaiso overexpressing mice (Kaiso Tg) exhibited chronic intestinal inflammation and had increased numbers of all three secretory cell types, hinting that Kaiso might regulate Notch signaling in the gut. However, Kaiso’s precise role in Notch signaling and whether the Kaiso Tg secretory cell fate phenotype was linked to Kaiso-induced inflammation had yet to be elucidated.MethodsIntestines from 3-month old Non-transgenic and Kaiso Tg mice were “Swiss” rolled and analysed for the expression of Notch1, Dll-1, Jagged-1, and secretory cell markers by immunohistochemistry and immunofluorescence. To evaluate inflammation, morphological analyses and myeloperoxidase assays were performed on intestines from 3-month old Kaiso Tg and control mice. Notch1, Dll-1 and Jagged-1 expression were also assessed in stable Kaiso-depleted colon cancer cells and isolated intestinal epithelial cells using real time PCR and western blotting. To assess Kaiso binding to the DLL1, JAG1 and NOTCH1 promoter regions, chromatin immunoprecipitation was performed on three colon cancer cell lines.ResultsHere we demonstrate that Kaiso promotes secretory cell hyperplasia independently of Kaiso-induced inflammation. Moreover, Kaiso regulates several components of the Notch signaling pathway in intestinal cells, namely, Dll-1, Jagged-1 and Notch1. Notably, we found that in Kaiso Tg mice intestines, Notch1 and Dll-1 expression are significantly reduced while Jagged-1 expression is increased. Chromatin immunoprecipitation experiments revealed that Kaiso associates with the DLL1 and JAG1 promoter regions in a methylation-dependent manner in colon carcinoma cell lines, suggesting that these Notch ligands are putative Kaiso target genes.ConclusionHere, we provide evidence that Kaiso’s effects on intestinal secretory cell fates precede the development of intestinal inflammation in Kaiso Tg mice. We also demonstrate that Kaiso inhibits the expression of Dll-1, which likely contributes to the secretory cell phenotype observed in our transgenic mice. In contrast, Kaiso promotes Jagged-1 expression, which may have implications in Notch-mediated colon cancer progression.Electronic supplementary materialThe online version of this article (doi:10.1186/s12964-017-0178-x) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 79%

Kaiso differentially regulates components of the Notch signaling pathway in intestinal cells

et al. 2017

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“…22 Furthermore, knocking out RBP-J in the intestinal epithelium in vivo leads to goblet cell hyperplasia, increased permeability, and chronic colitis characterized by Th17 cells. 23 Thus, g-secretase inhibitors and the epithelial RBP-J knockout mouse model have the same end point as both exhibit goblet cell abnormalities. To avoid tipping epithelial cell differentiation towards the goblet cell phenotype, we aimed at globally downregulating Notch-1 expression.…”

Section: Discussionmentioning

confidence: 97%

Mouse and human Notch-1 regulate mucosal immune responses

et al. 2014

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The Notch-1 signaling pathway is responsible for homeostatic tight junction expression in vitro, and promotes barrier function in vivo in the RAG1-adoptive transfer model of colitis. In this study, we sought to determine the role of colonic Notch-1 in the lymphoepithelial crosstalk in health and disease. We utilized in vivo and in vitro knockdown to target the expression of Notch-1. We identified that epithelial Notch-1 is required for appropriate activation of intestinal epithelial cells at steady state and upon inflammatory stimulus. Notch-1 expression modulates mucosal chemokine and cytokine secretion, and FoxP3 and effector T-cell responses. We showed that epithelial Notch-1 controls the immune function of the epithelium through crosstalk with the nuclear factor-κB (NF-κB)/mitogen-activated protein kinase (MAPK) pathways that, in turn, elicits T-cell responses. Overall, epithelial Notch-1 bridges innate and adaptive immunity in the gut. Our findings highlight an indispensable role for Notch-1-mediated signaling in the intricate epithelial-immune crosstalk, and validate that epithelial Notch-1 is necessary and sufficient to support protective epithelial proinflammatory responses.

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“…There are numerous studies demonstrating that Notch1 regulates the intestinal barrier. (Turgeon et al, 2013, Mathern et al, 2014, Min et al, 2014, Obata et al, 2012, Dahan et al, 2011) siRNA knockdown of Notch1 in vitro (i.e., Caco-2 cells) is associated with impaired intestinal cell monolayer integrity and decreased expression of the tight junction protein occludin. (Dahan et al, 2011) Downregulation of Notch1 in vivo is associated with gut leakiness and alterations in tight junction protein expression and colon inflammation.…”

Section: Discussionmentioning

confidence: 99%

“…(Mathern et al, 2014, Dahan et al, 2011, Min et al, 2014) Notch1 cleavage product NICD pairs with the transcription factor Rbpj in mice to activate canonical Notch1 target genes and in vivo knockout of Rbpj in mice causes intestinal hyperpermeability and colitis. (Obata et al, 2012) A recent study in diabetic mice showed that decreased NICD/Hes1 expression resulted in intestinal hyperpermeability. (Min et al, 2014) Significantly, a rectal enema containing siRNA to Notch1 administered to mice reduces colonic stem cell Notch1 expression and results in colonic hyperpermeability.…”

Section: Discussionmentioning

confidence: 99%

Alcohol Feeding in Mice Promotes Colonic Hyperpermeability and Changes in Colonic Organoid Stem Cell Fate

Forsyth

Shaikh

Bishehsari

et al. 2017

Alcoholism Clin & Exp Res

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Background Alcohol increases intestinal permeability to pro-inflammatory microbial products that promote liver disease, even after a period of sobriety. We sought to test the hypothesis that alcohol affects intestinal stem cells using an in vivo model and ex vivo organoids generated from jejunum and colon from mice fed chronic alcohol. Methods Mice were fed a control or an alcohol diet. Intestinal permeability, liver steatosis/inflammation, and stool short chain fatty acids (SCFA) were measured. Jejunum and colon organoids and tissue were stained for stem cell, cell lineage, and apical junction markers with assessment of mRNA by PCR and RNA-seq. ChIP-PCR analysis was carried out for Notch1 using an antibody specific for acetylated histone 3. Results Alcohol-fed mice exhibited colonic (but not small intestinal) hyperpermeability, steatohepatitis and decreased butyrate/total SCFA ratio in stool. Stem cell, cell lineage, and apical junction marker staining in tissue or organoids from jejunum tissue were not impacted by alcohol. Only chromogranin A (Chga) was increased in jejunum organoids by qPCR. However, colon tissue and organoid staining exhibited an alcohol-induced significant decrease in cytokeratin 20+ (Krt20) absorptive lineage enterocytes, a decrease in occludin and E-cadherin apical junction proteins, an increase in Chga, and an increase in the Lgr5 stem cell marker. qPCR revealed an alcohol-induced decrease in colon organoid and tissue Notch1, Hes1, and Krt20 and increased Chga, supporting an alteration in stem cell fate due to decreased Notch1 expression. Colon tissue ChIP-PCR revealed alcohol feeding suppressed Notch1 mRNA expression (via deacetylation of histone H3) and decreased Notch1 tissue staining. Conclusions Data support a model for alcohol-induced colonic hyperpermeability via epigenetic effects on Notch1, and thus Hes1, suppression through a mechanism involving histone H3 deacetylation at the Notch1 locus. This decreased enterocyte and increased enteroendocrine cell colonic stem cell fate and decreased apical junctional proteins leading to hyperpermeability.

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Epithelial Cell-Intrinsic Notch Signaling Plays an Essential Role in the Maintenance of Gut Immune Homeostasis

Cited by 63 publications

References 39 publications

Kaiso differentially regulates components of the Notch signaling pathway in intestinal cells

Kaiso differentially regulates components of the Notch signaling pathway in intestinal cells

Mouse and human Notch-1 regulate mucosal immune responses

Alcohol Feeding in Mice Promotes Colonic Hyperpermeability and Changes in Colonic Organoid Stem Cell Fate

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