The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe

ABSTRACT Pluripotent stem-cell-derived human intestinal organoids (HIOs) are three-dimensional, multicellular structures that model a naive intestinal epithelium in an in vitro system. Several published reports have investigated the use of HIOs to study host-microbe interactions. We recently demonst...

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Autores principales:	Madeline R. Barron, Roberto J. Cieza, David R. Hill, Sha Huang, Veda K. Yadagiri, Jason R. Spence, Vincent B. Young
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	RpoS colonization intestine organoid stress Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/3f9364e1354942f18694e094b73d3b36
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spelling	oai:doaj.org-article:3f9364e1354942f18694e094b73d3b362021-11-15T15:31:12ZThe Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe10.1128/mSphere.00777-202379-5042https://doaj.org/article/3f9364e1354942f18694e094b73d3b362020-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00777-20https://doaj.org/toc/2379-5042ABSTRACT Pluripotent stem-cell-derived human intestinal organoids (HIOs) are three-dimensional, multicellular structures that model a naive intestinal epithelium in an in vitro system. Several published reports have investigated the use of HIOs to study host-microbe interactions. We recently demonstrated that microinjection of the nonpathogenic Escherichia coli strain ECOR2 into HIOs induced morphological and functional maturation of the HIO epithelium, including increased secretion of mucins and cationic antimicrobial peptides. In the current work, we use ECOR2 as a biological probe to further characterize the environment present in the HIO lumen. We generated an isogenic mutant in the general stress response sigma factor RpoS and employed this mutant to compare challenges faced by a bacterium during colonization of the HIO lumen relative to the germ-free mouse intestine. We demonstrate that the loss of RpoS significantly decreases the ability of ECOR2 to colonize HIOs, although it does not prevent colonization of germ-free mice. These results indicate that the HIO lumen is a more restrictive environment to E. coli than the germ-free mouse intestine, thus increasing our understanding of the HIO model system as it pertains to studying the establishment of intestinal host-microbe symbioses. IMPORTANCE Technological advancements have driven and will continue to drive the adoption of organotypic systems for investigating host-microbe interactions within the human intestinal ecosystem. Using E. coli deficient in the RpoS-mediated general stress response, we demonstrate that the type or severity of microbial stressors within the HIO lumen is more restrictive than those of the in vivo environment of the germ-free mouse gut. This study provides important insight into the nature of the HIO microenvironment from a microbiological standpoint.Madeline R. BarronRoberto J. CiezaDavid R. HillSha HuangVeda K. YadagiriJason R. SpenceVincent B. YoungAmerican Society for MicrobiologyarticleRpoScolonizationintestineorganoidstressMicrobiologyQR1-502ENmSphere, Vol 5, Iss 6 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	RpoS colonization intestine organoid stress Microbiology QR1-502
spellingShingle	RpoS colonization intestine organoid stress Microbiology QR1-502 Madeline R. Barron Roberto J. Cieza David R. Hill Sha Huang Veda K. Yadagiri Jason R. Spence Vincent B. Young The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe
description	ABSTRACT Pluripotent stem-cell-derived human intestinal organoids (HIOs) are three-dimensional, multicellular structures that model a naive intestinal epithelium in an in vitro system. Several published reports have investigated the use of HIOs to study host-microbe interactions. We recently demonstrated that microinjection of the nonpathogenic Escherichia coli strain ECOR2 into HIOs induced morphological and functional maturation of the HIO epithelium, including increased secretion of mucins and cationic antimicrobial peptides. In the current work, we use ECOR2 as a biological probe to further characterize the environment present in the HIO lumen. We generated an isogenic mutant in the general stress response sigma factor RpoS and employed this mutant to compare challenges faced by a bacterium during colonization of the HIO lumen relative to the germ-free mouse intestine. We demonstrate that the loss of RpoS significantly decreases the ability of ECOR2 to colonize HIOs, although it does not prevent colonization of germ-free mice. These results indicate that the HIO lumen is a more restrictive environment to E. coli than the germ-free mouse intestine, thus increasing our understanding of the HIO model system as it pertains to studying the establishment of intestinal host-microbe symbioses. IMPORTANCE Technological advancements have driven and will continue to drive the adoption of organotypic systems for investigating host-microbe interactions within the human intestinal ecosystem. Using E. coli deficient in the RpoS-mediated general stress response, we demonstrate that the type or severity of microbial stressors within the HIO lumen is more restrictive than those of the in vivo environment of the germ-free mouse gut. This study provides important insight into the nature of the HIO microenvironment from a microbiological standpoint.
format	article
author	Madeline R. Barron Roberto J. Cieza David R. Hill Sha Huang Veda K. Yadagiri Jason R. Spence Vincent B. Young
author_facet	Madeline R. Barron Roberto J. Cieza David R. Hill Sha Huang Veda K. Yadagiri Jason R. Spence Vincent B. Young
author_sort	Madeline R. Barron
title	The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe
title_short	The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe
title_full	The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe
title_fullStr	The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe
title_full_unstemmed	The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe
title_sort	lumen of human intestinal organoids poses greater stress to bacteria compared to the germ-free mouse intestine: <named-content content-type="genus-species">escherichia coli</named-content> deficient in rpos as a colonization probe
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/3f9364e1354942f18694e094b73d3b36
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The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: <named-content content-type="genus-species">Escherichia coli</named-content> Deficient in RpoS as a Colonization Probe

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