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https://www.readbyqxmd.com/read/28090567/the-rna-polymerase-iii-subunit-polr3b-is-required-for-the-maintenance-of-small-intestinal-crypts-in-mice
#1
Julia Kieckhaefer, Sabina Lukovac, Diana Z Ye, Dolim Lee, Danielle J Beetler, Michael Pack, Klaus H Kaestner
BACKGROUND & AIMS: The continuously self-renewing mammalian intestinal epithelium, with high cellular turnover, depends on adequate protein synthesis for its proliferative capacity. RNA polymerase III activity is closely related to cellular growth and proliferation. Here, we studied the role of Polr3b, a large RNA polymerase III subunit, in the mammalian intestinal epithelium. METHODS: We derived mice with an intestinal epithelium-specific hypomorphic mutation of the Polr3b gene, using VillinCre-mediated gene ablation...
November 2016: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/28090566/large-animal-models-the-key-to-translational-discovery-in-digestive-disease-research
#2
Amanda Ziegler, Liara Gonzalez, Anthony Blikslager
Gastrointestinal disease is a prevalent cause of morbidity and mortality and the use of animal models have been instrumental in studying mechanisms of digestive pathophysiology. As investigators attempt to translate the wealth of basic science information developed from rodent, models, large animal models provide a number of translational advantages. The pig, in particular, is arguably one of the most powerful models of human organ systems, including the gastrointestinal tract. The pig has provided important tools and insight into intestinal ischemia/reperfusion injury, intestinal mucosal repair, as well as new insights into esophageal injury and repair...
November 2016: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/28069942/a-paradox-of-transcriptional-and-functional-innate-interferon-responses-of-human-intestinal-enteroids-to-enteric-virus-infection
#3
Kapil Saxena, Lukas M Simon, Xi-Lei Zeng, Sarah E Blutt, Sue E Crawford, Narayan P Sastri, Umesh C Karandikar, Nadim J Ajami, Nicholas C Zachos, Olga Kovbasnjuk, Mark Donowitz, Margaret E Conner, Chad A Shaw, Mary K Estes
The intestinal epithelium can limit enteric pathogens by producing antiviral cytokines, such as IFNs. Type I IFN (IFN-α/β) and type III IFN (IFN-λ) function at the epithelial level, and their respective efficacies depend on the specific pathogen and site of infection. However, the roles of type I and type III IFN in restricting human enteric viruses are poorly characterized as a result of the difficulties in cultivating these viruses in vitro and directly obtaining control and infected small intestinal human tissue...
January 9, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28053090/functional-transcriptomics-in-diverse-intestinal-epithelial-cell-types-reveals-robust-microrna-sensitivity-in-intestinal-stem-cells-to-microbial-status
#4
Bailey C E Peck, Amanda T Mah, Wendy A Pitman, Shengli Ding, P Kay Lund, Praveen Sethupathy
Gut microbiota play an important role in regulating the development of the host immune system, metabolic rate, and at times, disease pathogenesis. The factors and mechanisms that mediate interactions between microbiota and the intestinal epithelium are not fully understood. We provide novel evidence that microbiota may control intestinal epithelial stem cell (IESC) proliferation in part through microRNAs (miRNAs). We demonstrate that miRNA profiles differ dramatically across functionally distinct cell types of the mouse jejunal intestinal epithelium and that miRNAs respond to microbiota in a highly cell-type specific manner...
January 4, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28039407/cryptosporidium-parvum-infection-attenuates-the-ex%C3%A2-vivo-propagation-of-murine-intestinal-enteroids
#5
Xin-Tian Zhang, Ai-Yu Gong, Yang Wang, Xiqiang Chen, Sheng-Yau S Lim, Courtney E Dolata, Xian-Ming Chen
Cryptosporidium, a ubiquitous coccidian protozoan parasite that infects the gastrointestinal epithelium and other mucosal surfaces, is an important opportunistic pathogen for immunocompromised individuals and a common cause of diarrhea in young children in the developing countries. One of the pathological hallmarks of intestinal cryptosporidiosis is villous atrophy, which results in a shorter height of intestinal villi. Here, we investigated the effects of Cryptosporidium infection on intestinal epithelial growth, using an ex vivo model of intestinal cryptosporidiosis employing enteroids from mice...
December 2016: Physiological Reports
https://www.readbyqxmd.com/read/27871064/benzopyrimido-pyrrolo-oxazine-dione-cftr-inhibitor-r-bpo-27-for-anti-secretory-therapy-of-diarrheas-caused-by-bacterial-enterotoxins
#6
Onur Cil, Puay-Wah Phuan, Anne Marie Gillespie, Sujin Lee, Lukmanee Tradtrantip, Jianyi Yin, Ming Tse, Nicholas C Zachos, Ruxian Lin, Mark Donowitz, Alan S Verkman
Secretory diarrheas caused by bacterial enterotoxins, including cholera and traveler's diarrhea, remain a major global health problem. Inappropriate activation of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel occurs in these diarrheas. We previously reported that the benzopyrimido-pyrrolo-oxazinedione (R)-BPO-27 inhibits CFTR chloride conductance with low-nanomolar potency. Here, we demonstrate using experimental mouse models and human enterocyte cultures the potential utility of (R)-BPO-27 for treatment of secretory diarrheas caused by cholera and Escherichia coli enterotoxins...
November 8, 2016: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/27867006/intercellular-coupling-of-the-cell-cycle-and-circadian-clock-in-adult-stem-cell-culture
#7
Toru Matsu-Ura, Andrey Dovzhenok, Eitaro Aihara, Jill Rood, Hung Le, Yan Ren, Andrew E Rosselot, Tongli Zhang, Choogon Lee, Karl Obrietan, Marshall H Montrose, Sookkyung Lim, Sean R Moore, Christian I Hong
Circadian clock-gated cell division cycles are observed from cyanobacteria to mammals via intracellular molecular connections between these two oscillators. Here we demonstrate WNT-mediated intercellular coupling between the cell cycle and circadian clock in 3D murine intestinal organoids (enteroids). The circadian clock gates a population of cells with heterogeneous cell-cycle times that emerge as 12-hr synchronized cell division cycles. Remarkably, we observe reduced-amplitude oscillations of circadian rhythms in intestinal stem cells and progenitor cells, indicating an intercellular signal arising from differentiated cells governing circadian clock-dependent synchronized cell division cycles...
December 1, 2016: Molecular Cell
https://www.readbyqxmd.com/read/27815136/phenylquinoxalinone-cftr-activator-as-potential-prosecretory-therapy-for-constipation
#8
Onur Cil, Puay-Wah Phuan, Jung-Ho Son, Jie S Zhu, Colton K Ku, Niloufar Akhavan Tabib, Andrew P Teuthorn, Loretta Ferrera, Nicholas C Zachos, Ruxian Lin, Luis J V Galietta, Mark Donowitz, Mark J Kurth, A S Verkman
Constipation is a common condition for which current treatments can have limited efficacy. By high-throughput screening, we recently identified a phenylquinoxalinone activator of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel that stimulated intestinal fluid secretion and normalized stool output in a mouse model of opioid-induced constipation (Cil et al Cell Mol Gastroenterol Hepatol 2:317-327, 2016). Here, we report phenylquinoxalinone structure-activity analysis, mechanism of action, animal efficacy data in acute and chronic models of constipation, and functional data in ex vivo primary cultured human enterocytes...
October 15, 2016: Translational Research: the Journal of Laboratory and Clinical Medicine
https://www.readbyqxmd.com/read/27791005/single-cell-lineage-tracing-reveals-a-role-for-tgf%C3%AE-r2-in-intestinal-stem-cell-dynamics-and-differentiation
#9
Jared M Fischer, Peter P Calabrese, Ashleigh J Miller, Nina M Muñoz, William M Grady, Darryl Shibata, R Michael Liskay
Intestinal stem cells (ISCs) are maintained by a niche mechanism, in which multiple ISCs undergo differential fates where a single ISC clone ultimately occupies the niche. Importantly, mutations continually accumulate within ISCs creating a potential competitive niche environment. Here we use single cell lineage tracing following stochastic transforming growth factor β receptor 2 (TgfβR2) mutation to show cell autonomous effects of TgfβR2 loss on ISC clonal dynamics and differentiation. Specifically, TgfβR2 mutation in ISCs increased clone survival while lengthening times to monoclonality, suggesting that Tgfβ signaling controls both ISC clone extinction and expansion, independent of proliferation...
October 25, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27787775/the-three-dimensional-culture-of-epithelial-organoids-derived-from-embryonic-chicken-intestine
#10
Malgorzata Pierzchalska, Malgorzata Panek, Malgorzata Czyrnek, Maja Grabacka
The intestinal epithelium isolated from chicken embryos in last 3 days of development can be used to establish the 3D culture of intestinal organoids. When fragments of epithelial tissue released by incubation with EGTA (2.5 mM, 2 h) are embedded in Matrigel matrix on cell culture inserts the formation of empty spheres covered by epithelial cells is observed in first 24 h of culture. The growth and survival of organoids are supported by the addition of R-spondin 1, Noggin, and prostaglandin E2 to the culture medium...
October 28, 2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27685631/mesenchymal-stem-cells-stimulate-intestinal-stem-cells-to-repair-radiation-induced-intestinal-injury
#11
Wei Gong, Mengzheng Guo, Zhibo Han, Yan Wang, Ping Yang, Chang Xu, Qin Wang, Liqing Du, Qian Li, Hui Zhao, Feiyue Fan, Qiang Liu
The loss of stem cells residing in the base of the intestinal crypt has a key role in radiation-induced intestinal injury. In particular, Lgr5(+) intestinal stem cells (ISCs) are indispensable for intestinal regeneration following exposure to radiation. Mesenchymal stem cells (MSCs) have previously been shown to improve intestinal epithelial repair in a mouse model of radiation injury, and, therefore, it was hypothesized that this protective effect is related to Lgr5(+) ISCs. In this study, it was found that, following exposure to radiation, transplantation of MSCs improved the survival of the mice, ameliorated intestinal injury and increased the number of regenerating crypts...
2016: Cell Death & Disease
https://www.readbyqxmd.com/read/27677718/human-mini-guts-new-insights-into-intestinal-physiology-and-host-pathogen-interactions
#12
REVIEW
Julie G In, Jennifer Foulke-Abel, Mary K Estes, Nicholas C Zachos, Olga Kovbasnjuk, Mark Donowitz
The development of indefinitely propagating human 'mini-guts' has led to a rapid advance in gastrointestinal research related to transport physiology, developmental biology, pharmacology, and pathophysiology. These mini-guts, also called enteroids or colonoids, are derived from LGR5(+) intestinal stem cells isolated from the small intestine or colon. Addition of WNT3A and other growth factors promotes stemness and results in viable, physiologically functional human intestinal or colonic cultures that develop a crypt-villus axis and can be differentiated into all intestinal epithelial cell types...
November 2016: Nature Reviews. Gastroenterology & Hepatology
https://www.readbyqxmd.com/read/27632431/intestinal-organoids-new-frontiers-in-the-study-of-intestinal-disease-and-physiology
#13
Thomas E Wallach, James R Bayrer
The development of sustainable intestinal organoid cell culture has emerged as a new modality for the study of intestinal function and cellular processes. Organoid culture is providing a new testbed for therapeutic research and development. Intestinal organoids, self-renewing 3-dimensional structures comprised intestinal stem cells and their differentiated epithelial progeny allow for more facile and robust exploration of cellular activity, cell organization and structure, genetic manipulation, and vastly more physiologic modeling of intestinal response to stimuli as compared to traditional 2-dimensional cell line cultures...
February 2017: Journal of Pediatric Gastroenterology and Nutrition
https://www.readbyqxmd.com/read/27562956/replication-of-human-noroviruses-in-stem-cell-derived-human-enteroids
#14
Khalil Ettayebi, Sue E Crawford, Kosuke Murakami, James R Broughman, Umesh Karandikar, Victoria R Tenge, Frederick H Neill, Sarah E Blutt, Xi-Lei Zeng, Lin Qu, Baijun Kou, Antone R Opekun, Douglas Burrin, David Y Graham, Sasirekha Ramani, Robert L Atmar, Mary K Estes
The major barrier to research and development of effective interventions for human noroviruses (HuNoVs) has been the lack of a robust and reproducible in vitro cultivation system. HuNoVs are the leading cause of gastroenteritis worldwide. We report the successful cultivation of multiple HuNoV strains in enterocytes in stem cell-derived, nontransformed human intestinal enteroid monolayer cultures. Bile, a critical factor of the intestinal milieu, is required for strain-dependent HuNoV replication. Lack of appropriate histoblood group antigen expression in intestinal cells restricts virus replication, and infectivity is abrogated by inactivation (e...
September 23, 2016: Science
https://www.readbyqxmd.com/read/27561422/the-transcriptional-and-splicing-landscape-of-intestinal-organoids-undergoing-nutrient-starvation-or-endoplasmic-reticulum-stress
#15
Jessica Tsalikis, Qun Pan, Ivan Tattoli, Charles Maisonneuve, Benjamin J Blencowe, Dana J Philpott, Stephen E Girardin
BACKGROUND: The intestinal epithelium plays a critical role in nutrient absorption and innate immune defense. Recent studies showed that metabolic stress pathways, in particular the integrated stress response (ISR), control intestinal epithelial cell fate and function. Here, we used RNA-seq to analyze the global transcript level and alternative splicing responses of primary murine enteroids undergoing two distinct ISR-triggering stresses, endoplasmic reticulum (ER) stress and nutrient starvation...
2016: BMC Genomics
https://www.readbyqxmd.com/read/27546461/iron-uptake-via-dmt1-integrates-cell-cycle-with-jak-stat3-signaling-to-promote-colorectal-tumorigenesis
#16
Xiang Xue, Sadeesh K Ramakrishnan, Kevin Weisz, Daniel Triner, Liwei Xie, Durga Attili, Asha Pant, Balázs Győrffy, Mingkun Zhan, Christin Carter-Su, Karin M Hardiman, Thomas D Wang, Michael K Dame, James Varani, Dean Brenner, Eric R Fearon, Yatrik M Shah
Dietary iron intake and systemic iron balance are implicated in colorectal cancer (CRC) development, but the means by which iron contributes to CRC are unclear. Gene expression and functional studies demonstrated that the cellular iron importer, divalent metal transporter 1 (DMT1), is highly expressed in CRC through hypoxia-inducible factor 2α-dependent transcription. Colon-specific Dmt1 disruption resulted in a tumor-selective inhibitory effect of proliferation in mouse colon tumor models. Proteomic and genomic analyses identified an iron-regulated signaling axis mediated by cyclin-dependent kinase 1 (CDK1), JAK1, and STAT3 in CRC progression...
September 13, 2016: Cell Metabolism
https://www.readbyqxmd.com/read/27451883/production-of-tissue-engineered-intestine-from-expanded-enteroids
#17
Barrett P Cromeens, Yanchun Liu, Johnathan Stathopoulos, Yijie Wang, Jed Johnson, Gail E Besner
BACKGROUND: Short bowel syndrome is a life-threatening condition with few solutions. Tissue-engineered intestine (TEI) is a potential treatment, but donor intestine is a limiting factor. Expanded epithelial surrogates termed enteroids may serve as a potential donor source. MATERIALS AND METHODS: To produce TEI from enteroids, crypts were harvested from mice and enteroid cultures established. Enteroids were seeded onto polymer scaffolds using Matrigel or culture medium and implanted in immunosuppressed mice for 4 wk...
July 2016: Journal of Surgical Research
https://www.readbyqxmd.com/read/27413168/tnf-%C3%AE-augments-rankl-dependent-intestinal-m-cell-differentiation-in-enteroid-cultures
#18
Megan B Wood, Daniel Rios, Ifor R Williams
Microfold (M) cells are phagocytic intestinal epithelial cells in the follicle-associated epithelium of Peyer's patches that transport particulate antigens from the gut lumen into the subepithelial dome. Differentiation of M cells from epithelial stem cells in intestinal crypts requires the cytokine receptor activator of NF-κB ligand (RANKL) and the transcription factor Spi-B. We used three-dimensional enteroid cultures established with small intestinal crypts from mice as a model system to investigate signaling pathways involved in M cell differentiation and the influence of other cytokines on RANKL-induced M cell differentiation...
September 1, 2016: American Journal of Physiology. Cell Physiology
https://www.readbyqxmd.com/read/27353252/an-lgg-derived-protein-promotes-iga-production-through-upregulation-of-april-expression-in-intestinal-epithelial-cells
#19
Y Wang, L Liu, D J Moore, X Shen, R M Peek, S A Acra, H Li, X Ren, D B Polk, F Yan
p40, a Lactobacillus rhamnosus GG (LGG)-derived protein, transactivates epidermal growth factor receptor (EGFR) in intestinal epithelial cells, leading to amelioration of intestinal injury and inflammation. To elucidate mechanisms by which p40 regulates mucosal immunity to prevent inflammation, this study aimed to determine the effects and mechanisms of p40 on regulation of a proliferation-inducing ligand (APRIL) expression in intestinal epithelial cells for promoting immunoglobulin A (IgA) production. p40 upregulated April gene expression and protein production in mouse small intestine epithelial (MSIE) cells, which were inhibited by blocking EGFR expression and kinase activity...
June 29, 2016: Mucosal Immunology
https://www.readbyqxmd.com/read/27162272/blood-group-o-dependent-cellular-responses-to-cholera-toxin-parallel-clinical-and-epidemiological-links-to-severe-cholera
#20
F Matthew Kuhlmann, Srikanth Santhanam, Pardeep Kumar, Qingwei Luo, Matthew A Ciorba, James M Fleckenstein
Because O blood group has been associated with more severe cholera infections, it has been hypothesized that cholera toxin (CT) may bind non-O blood group antigens of the intestinal mucosae, thereby preventing efficient interaction with target GM1 gangliosides required for uptake of the toxin and activation of cyclic adenosine monophosphate (cAMP) signaling in target epithelia. Herein, we show that after exposure to CT, human enteroids expressing O blood group exhibited marked increase in cAMP relative to cells derived from blood group A individuals...
August 3, 2016: American Journal of Tropical Medicine and Hygiene
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