Microbial Metabolites Orchestrate a Distinct Multi-Tiered Regulatory Network in the Intestinal Epithelium That Directs P-Glycoprotein Expression
ABSTRACT P-glycoprotein (P-gp) is a key component of the intestinal epithelium playing a pivotal role in removal of toxins and efflux of endocannabinoids to prevent excessive inflammation and sustain homeostasis. Recent studies revealed butyrate and secondary bile acids, produced by the intestinal m...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2022-08-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.01993-22 |
| _version_ | 1818473367759486976 |
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| author | Sage E. Foley Michael J. Dente Xuqiu Lei Benjamin F. Sallis Ethan B. Loew Mario Meza-Segura Katherine A. Fitzgerald Beth A. McCormick |
| author_facet | Sage E. Foley Michael J. Dente Xuqiu Lei Benjamin F. Sallis Ethan B. Loew Mario Meza-Segura Katherine A. Fitzgerald Beth A. McCormick |
| author_sort | Sage E. Foley |
| collection | DOAJ |
| description | ABSTRACT P-glycoprotein (P-gp) is a key component of the intestinal epithelium playing a pivotal role in removal of toxins and efflux of endocannabinoids to prevent excessive inflammation and sustain homeostasis. Recent studies revealed butyrate and secondary bile acids, produced by the intestinal microbiome, potentiate the induction of functional P-gp expression. We now aim to determine the molecular mechanism by which this functional microbiome output regulates P-gp. RNA sequencing of intestinal epithelial cells responding to butyrate and secondary bile acids in combination discovered a unique transcriptional program involving multiple pathways that converge on P-gp induction. Using shRNA knockdown and CRISPR/Cas9 knockout cell lines, as well as mouse models, we confirmed the RNA sequencing findings and discovered a role for intestinal HNF4α in P-gp regulation. These findings shed light on a sophisticated signaling network directed by intestinal microbial metabolites that orchestrate P-gp expression and highlight unappreciated connections between multiple pathways linked to colonic health. IMPORTANCE Preventing aberrant inflammation is essential to maintaining homeostasis in the mammalian intestine. Although P-glycoprotein (P-gp) expression in the intestine is critical for protecting the intestinal epithelium from toxins and damage due to neutrophil infiltration, its regulation in the intestine is poorly understood. Findings presented in our current study have now uncovered a sophisticated and heretofore unappreciated intracellular signaling network or “reactome” directed by intestinal microbial metabolites that orchestrate regulation of P-gp. Not only do we confirm the role of histone deacetylases (HDAC) inhibition and nuclear receptor activation in P-gp induction by butyrate and bile acids, but we also discovered new signaling pathways and transcription factors that are uniquely activated in response to the combination of microbial metabolites. Such findings shed new light into a multi-tiered network that maintains P-gp expression in the intestine in the context of the fluctuating commensal microbiome, to sustain a homeostatic tone in the absence of infection or insult. |
| first_indexed | 2024-04-14T04:21:58Z |
| format | Article |
| id | doaj.art-6f9a0174e68b46aab65cc51c0cd152b5 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-04-14T04:21:58Z |
| publishDate | 2022-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-6f9a0174e68b46aab65cc51c0cd152b52022-12-22T02:12:28ZengAmerican Society for MicrobiologymBio2150-75112022-08-0113410.1128/mbio.01993-22Microbial Metabolites Orchestrate a Distinct Multi-Tiered Regulatory Network in the Intestinal Epithelium That Directs P-Glycoprotein ExpressionSage E. Foley0Michael J. Dente1Xuqiu Lei2Benjamin F. Sallis3Ethan B. Loew4Mario Meza-Segura5Katherine A. Fitzgerald6Beth A. McCormick7Department of Microbiology and Physiological Systems, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USAT.H. Chan School of Medicine, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USAProgram in Innate Immunity, Department of Medicine, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USADepartment of Microbiology and Physiological Systems, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USADepartment of Microbiology and Physiological Systems, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USADepartment of Microbiology and Physiological Systems, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USAProgram in Innate Immunity, Department of Medicine, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USADepartment of Microbiology and Physiological Systems, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USAABSTRACT P-glycoprotein (P-gp) is a key component of the intestinal epithelium playing a pivotal role in removal of toxins and efflux of endocannabinoids to prevent excessive inflammation and sustain homeostasis. Recent studies revealed butyrate and secondary bile acids, produced by the intestinal microbiome, potentiate the induction of functional P-gp expression. We now aim to determine the molecular mechanism by which this functional microbiome output regulates P-gp. RNA sequencing of intestinal epithelial cells responding to butyrate and secondary bile acids in combination discovered a unique transcriptional program involving multiple pathways that converge on P-gp induction. Using shRNA knockdown and CRISPR/Cas9 knockout cell lines, as well as mouse models, we confirmed the RNA sequencing findings and discovered a role for intestinal HNF4α in P-gp regulation. These findings shed light on a sophisticated signaling network directed by intestinal microbial metabolites that orchestrate P-gp expression and highlight unappreciated connections between multiple pathways linked to colonic health. IMPORTANCE Preventing aberrant inflammation is essential to maintaining homeostasis in the mammalian intestine. Although P-glycoprotein (P-gp) expression in the intestine is critical for protecting the intestinal epithelium from toxins and damage due to neutrophil infiltration, its regulation in the intestine is poorly understood. Findings presented in our current study have now uncovered a sophisticated and heretofore unappreciated intracellular signaling network or “reactome” directed by intestinal microbial metabolites that orchestrate regulation of P-gp. Not only do we confirm the role of histone deacetylases (HDAC) inhibition and nuclear receptor activation in P-gp induction by butyrate and bile acids, but we also discovered new signaling pathways and transcription factors that are uniquely activated in response to the combination of microbial metabolites. Such findings shed new light into a multi-tiered network that maintains P-gp expression in the intestine in the context of the fluctuating commensal microbiome, to sustain a homeostatic tone in the absence of infection or insult.https://journals.asm.org/doi/10.1128/mbio.01993-22P-glycoproteinmulti-drug resistance transportermicrobiomemetabolitesbutyrateshort-chain fatty acids |
| spellingShingle | Sage E. Foley Michael J. Dente Xuqiu Lei Benjamin F. Sallis Ethan B. Loew Mario Meza-Segura Katherine A. Fitzgerald Beth A. McCormick Microbial Metabolites Orchestrate a Distinct Multi-Tiered Regulatory Network in the Intestinal Epithelium That Directs P-Glycoprotein Expression mBio P-glycoprotein multi-drug resistance transporter microbiome metabolites butyrate short-chain fatty acids |
| title | Microbial Metabolites Orchestrate a Distinct Multi-Tiered Regulatory Network in the Intestinal Epithelium That Directs P-Glycoprotein Expression |
| title_full | Microbial Metabolites Orchestrate a Distinct Multi-Tiered Regulatory Network in the Intestinal Epithelium That Directs P-Glycoprotein Expression |
| title_fullStr | Microbial Metabolites Orchestrate a Distinct Multi-Tiered Regulatory Network in the Intestinal Epithelium That Directs P-Glycoprotein Expression |
| title_full_unstemmed | Microbial Metabolites Orchestrate a Distinct Multi-Tiered Regulatory Network in the Intestinal Epithelium That Directs P-Glycoprotein Expression |
| title_short | Microbial Metabolites Orchestrate a Distinct Multi-Tiered Regulatory Network in the Intestinal Epithelium That Directs P-Glycoprotein Expression |
| title_sort | microbial metabolites orchestrate a distinct multi tiered regulatory network in the intestinal epithelium that directs p glycoprotein expression |
| topic | P-glycoprotein multi-drug resistance transporter microbiome metabolites butyrate short-chain fatty acids |
| url | https://journals.asm.org/doi/10.1128/mbio.01993-22 |
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