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https://www.readbyqxmd.com/read/30111276/generation-of-renewable-mouse-intestinal-epithelial-cell-monolayers-and-organoids-for-functional-analyses
#1
Emily C Moorefield, R Eric Blue, Nancy L Quinney, Martina Gentzsch, Shengli Ding
BACKGROUND: Conditional reprogramming has enabled the development of long-lived, normal epithelial cell lines from mice and humans by in vitro culture with ROCK inhibitor on a feeder layer. We applied this technology to mouse small intestine to create 2D mouse intestinal epithelial monolayers (IEC monolayers) from genetic mouse models for functional analysis. RESULTS: IEC monolayers form epithelial colonies that proliferate on a feeder cell layer and are able to maintain their genotype over long-term passage...
August 15, 2018: BMC Cell Biology
https://www.readbyqxmd.com/read/30109253/epithelial-smad4-deletion-up-regulates-inflammation-and-promotes-inflammation-associated-cancer
#2
Anna L Means, Tanner J Freeman, Jing Zhu, Luke G Woodbury, Paula Marincola-Smith, Chao Wu, Anne R Meyer, Connie J Weaver, Chandrasekhar Padmanabhan, Hanbing An, Jinghuan Zi, Bronson C Wessinger, Rupesh Chaturvedi, Tasia D Brown, Natasha G Deane, Robert J Coffey, Keith T Wilson, J Joshua Smith, Charles L Sawyers, James R Goldenring, Sergey V Novitskiy, M Kay Washington, Chanjuan Shi, R Daniel Beauchamp
Background & Aims: Chronic inflammation is a predisposing condition for colorectal cancer. Many studies to date have focused on proinflammatory signaling pathways in the colon. Understanding the mechanisms that suppress inflammation, particularly in epithelial cells, is critical for developing therapeutic interventions. Here, we explored the roles of transforming growth factor β (TGFβ) family signaling through SMAD4 in colonic epithelial cells. Methods: The Smad4 gene was deleted specifically in adult murine intestinal epithelium...
2018: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/30105005/melatonin-improves-laying-performance-by-enhancing-intestinal-amino-acids-transport-in-hens
#3
Lijuan Liu, Siyu Zhang, Jiayang Bao, Xiaowen He, Danni Tong, Cong Chen, Qingxiang Ying, Qing Zhang, Caiqiao Zhang, Jian Li
The high concentration of melatonin (MEL) in the intestinal mucosa suggests that it has a special physiological function in intestine. In hens, previous studies have shown that MEL treatment promoted egg-laying performance. Considering the importance of amino acids (AA) for egg formation, we hypothesized that MEL may enhance the intestinal absorption of AA from the feed, thus promoting egg laying performance. In this study, we supplemented the hens with MEL for two consecutive weeks. The results showed that, compared with control group, feeding with 0...
2018: Frontiers in Endocrinology
https://www.readbyqxmd.com/read/30104275/nitazoxanide-inhibits-human-norovirus-replication-and-synergizes-with-ribavirin-by-activation-of-cellular-antiviral-response
#4
Wen Dang, Lei Xu, Buyun Ma, Sunrui Chen, Yuebang Yin, Kyeong-Ok Chang, Maikel P Peppelenbosch, Qiuwei Pan
Norovirus is the main cause of viral gastroenteritis worldwide. Although norovirus gastroenteritis is self-limiting in immunocompetent individuals, chronic infections with debilitating and life-threatening complications occur in immunocompromised patients. Nitazoxanide (NTZ) has been empirically used in the clinic and demonstrated effectiveness against norovirus gastroenteritis. In this study we aimed at uncovering the antiviral potential and mechanisms of NTZ and its active metabolite, tizoxanide (TIZ) using a human norovirus (HuNV) replicon...
August 13, 2018: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/30094477/adenylyl-cyclase-6-is-involved-in-the-hyposecretory-status-of-experimental-colitis
#5
Isabel Romero-Calvo, Borja Ocón, Reyes Gámez-Belmonte, Cristina Hernández-Chirlaque, Hugo R de Jonge, Marcel J Bijvelds, Olga Martínez-Augustin, Fermín Sánchez de Medina
One of the cardinal symptoms of intestinal inflammation is diarrhea. Acute intestinal inflammation is associated with inhibition of ion absorption and increased secretion, along with fluid leakage due to epithelial injury and changes in permeability. However, in the chronic situation, a downregulation of both absorptive and secretory transport has been reported. We investigated how experimental colitis reduces cAMP levels in intestinal epithelial cells through modulation of adenylyl cyclases (AC). Primary colonic epithelial cells obtained from rats with trinitrobenzenesulfonic acid colitis and non-colitic controls were analyzed for AC expression by RT-qPCR and Western blot, following a preliminary microarray analysis...
August 9, 2018: Pflügers Archiv: European Journal of Physiology
https://www.readbyqxmd.com/read/30072687/interleukin-2-induces-the-in-vitro-maturation-of-human-pluripotent-stem-cell-derived-intestinal-organoids
#6
Kwang Bo Jung, Hana Lee, Ye Seul Son, Mi-Ok Lee, Young-Dae Kim, Soo Jin Oh, Ohman Kwon, Sunwha Cho, Hyun-Soo Cho, Dae-Soo Kim, Jung-Hwa Oh, Matthias Zilbauer, Jeong-Ki Min, Cho-Rok Jung, Janghwan Kim, Mi-Young Son
Human pluripotent stem cell (hPSC)-derived intestinal organoids (hIOs) form 3D structures organized into crypt and villus domains, making them an excellent in vitro model system for studying human intestinal development and disease. However, hPSC-derived hIOs still require in vivo maturation to fully recapitulate adult intestine, with the mechanism of maturation remaining elusive. Here, we show that the co-culture with human T lymphocytes induce the in vitro maturation of hIOs, and identify STAT3-activating interleukin-2 (IL-2) as the major factor inducing maturation...
August 2, 2018: Nature Communications
https://www.readbyqxmd.com/read/30072255/transplantation-of-human-intestinal-organoids-into-the-mouse-mesentery-a-more-physiologic-and-anatomic-engraftment-site
#7
Alexander R Cortez, Holly M Poling, Nicole E Brown, Akaljot Singh, Maxime M Mahe, Michael A Helmrath
BACKGROUND: We previously described the development of human intestinal organoids from pluripotent stem cells, as well as their in vivo maturation when transplanted into the mouse kidney capsule. While sufficient for certain aspects of study, this model has limitations. Herein, we describe an alternative model of human intestinal organoids transplantation into the mouse mesentery. We hypothesize that efficient engraftment and marked differentiation of human intestinal organoids will be similar to our kidney model yet in a more anatomically appropriate location allowing for improved in vivo modeling...
July 30, 2018: Surgery
https://www.readbyqxmd.com/read/30060804/wnt-pcp-controls-spreading-of-wnt-%C3%AE-catenin-signals-by-cytonemes-in-vertebrates
#8
Benjamin Mattes, Yonglong Dang, Gediminas Greicius, Lilian Tamara Kaufmann, Benedikt Prunsche, Jakob Rosenbauer, Johannes Stegmaier, Ralf Mikut, Suat Özbek, Gerd Ulrich Nienhaus, Alexander Schug, David M Virshup, Steffen Scholpp
Signaling filopodia, termed cytonemes, are dynamic actin-based membrane structures that regulate the exchange of signaling molecules and their receptors within tissues. However, how cytoneme formation is regulated remains unclear. Here, we show that Wnt/planar cell polarity (PCP) autocrine signaling controls the emergence of cytonemes, and that cytonemes subsequently control paracrine Wnt/β-catenin signal activation. Upon binding of the Wnt family member Wnt8a, the receptor tyrosine kinase Ror2 becomes activated...
July 31, 2018: ELife
https://www.readbyqxmd.com/read/30055169/sox4-promotes-atoh1-independent-intestinal-secretory-differentiation-toward-tuft-and-enteroendocrine-fates
#9
Adam D Gracz, Leigh Ann Samsa, Matthew J Fordham, Danny C Trotier, Bailey Zwarycz, Yuan-Hung Lo, Katherine Bao, Joshua Starmer, Jesse R Raab, Noah F Shroyer, R Lee Reinhardt, Scott T Magness
BACKGROUND & AIMS: The intestinal epithelium is maintained by intestinal stem cells (ISCs), which produce post-mitotic absorptive and secretory epithelial cells. Initial fate specification toward enteroendocrine, goblet, and Paneth cell lineages requires the; transcription factor ATOH1, which regulates differentiation of the secretory cell lineage. However, less is known about the origin of tuft cells, which participate in type 2 immune; responses to parasite infections and appear to differentiate independently of ATOH1...
July 25, 2018: Gastroenterology
https://www.readbyqxmd.com/read/30039581/brief-summary-of-the-current-protocols-for-generating-intestinal-organoids
#10
REVIEW
Shizuka Miura, Atsushi Suzuki
The intestine has fundamental functions for the maintenance of homeostasis, including food digestion and nutrient/water absorption. Although the lumen of the intestine is always exposed to pathogens, intestinal epithelial cells form monolayer sheets that act as an epithelial barrier to prevent the invasion of pathogens. Thus, disruption of the intestinal epithelial barrier causes inflammatory bowel diseases. To investigate the details of these intractable intestinal diseases, it is necessary to analyze the characteristics of intestinal epithelial cells in vitro...
July 23, 2018: Development, Growth & Differentiation
https://www.readbyqxmd.com/read/30038251/enteroendocrine-cells-switch-hormone-expression-along-the-crypt-to-villus-bmp-signalling-gradient
#11
Joep Beumer, Benedetta Artegiani, Yorick Post, Frank Reimann, Fiona Gribble, Thuc Nghi Nguyen, Hongkui Zeng, Maaike Van den Born, Johan H Van Es, Hans Clevers
Enteroendocrine cells (EECs) control a wide range of physiological processes linked to metabolism1 . We show that EEC hormones are differentially expressed between crypts (for example, Glp1) and villi (for example, secretin). As demonstrated by single-cell mRNA sequencing using murine Lgr5+ cell-derived organoids, BMP4 signals alter the hormone expression profiles of individual EECs to resemble those found in the villus. Accordingly, BMP4 induces hormone switching of EECs migrating up the crypt-villus axis in vivo...
August 2018: Nature Cell Biology
https://www.readbyqxmd.com/read/30037999/a-population-of-gut-epithelial-enterochromaffin-cells-is-mechanosensitive-and-requires-piezo2-to-convert-force-into-serotonin-release
#12
Constanza Alcaino, Kaitlyn R Knutson, Anthony J Treichel, Gulcan Yildiz, Peter R Strege, David R Linden, Joyce H Li, Andrew B Leiter, Joseph H Szurszewski, Gianrico Farrugia, Arthur Beyder
Enterochromaffin (EC) cells constitute the largest population of intestinal epithelial enteroendocrine (EE) cells. EC cells are proposed to be specialized mechanosensory cells that release serotonin in response to epithelial forces, and thereby regulate intestinal fluid secretion. However, it is unknown whether EE and EC cells are directly mechanosensitive, and if so, what the molecular mechanism of their mechanosensitivity is. Consequently, the role of EE and EC cells in gastrointestinal mechanobiology is unclear...
August 7, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/30030066/r560s-a-class-ii-cftr-mutation-that-is-not-rescued-by-current-modulators
#13
Nikhil T Awatade, Sofia Ramalho, Iris A L Silva, Verónica Felício, Hugo M Botelho, Eyleen de Poel, Annelotte Vonk, Jeffrey M Beekman, Carlos M Farinha, Margarida D Amaral
BACKGROUND: New therapies modulating defective CFTR have started to hit the clinic and others are in trial or under development. The endeavour of drug discovery for CFTR protein rescue is however difficult one since over 2000 mutations have been reported. For most of these, especially the rare ones, the associated defects, the respective functional class and their responsiveness to available modulators are still unknown. Our aim here was to characterize the rare R560S mutation using patient-derived materials (rectal biopsies and intestinal organoids) from one CF individual homozygous for this mutation, in parallel with cellular models expressing R560S-CFTR and to assess the functional and biochemical responses to CFTR modulators...
July 18, 2018: Journal of Cystic Fibrosis: Official Journal of the European Cystic Fibrosis Society
https://www.readbyqxmd.com/read/30024920/modeling-apc-mutagenesis-and-familial-adenomatous-polyposis-using-human-ips-cells
#14
Cesar A Sommer, Amalia Capilla, Francisco J Molina-Estevez, Andreia Gianotti-Sommer, Nicholas Skvir, Ignacio Caballero, Sanjib Chowdhury, Gustavo Mostoslavsky
Mutations in the gene Adenomatous Polyposis Coli or APC appear in most sporadic cases of colorectal cancer and it is the most frequent mutation causing hereditary Familial Adenomatous Polyposis. The detailed molecular mechanism by which APC mutations predispose to the development of colorectal cancer is not completely understood. This is in part due to the lack of accessibility to appropriate models that recapitulate the early events associated with APC mediated intestinal transformation. We have established a novel platform utilizing human induced Pluripotent Stem cells or iPSC from normal or FAP-specific APC mutant individuals and evaluated the effect of the mutation in the cells before and after differentiation into intestinal organoids...
2018: PloS One
https://www.readbyqxmd.com/read/30022975/expression-and-lacking-internalization-of-the-cell-surface-receptors-of-clostridioides-difficile-toxin-b
#15
Dennis Schöttelndreier, Katrin Seeger, Guntram A Grassl, Markus R Winny, Robert Lindner, Harald Genth
Toxin-producing strains of Clostridioides difficile and Clostridium perfringens cause infections of the gastrointestinal tract in humans and ruminants, with the toxins being major virulence factors, essential for the infection, and responsible for the onset of severe symptoms. C. difficile toxin A (TcdA) and toxin B (TcdB), and the large cytotoxin (TpeL) from C. perfringens are single chain bacterial protein toxins with an AB-like toxin structure. The C-terminal delivery domain mediates cell entry of the N-terminal glycosyltransferase domain by receptor-mediated endocytosis...
2018: Frontiers in Microbiology
https://www.readbyqxmd.com/read/30017821/the-zinc-finger-transcription-factor-plagl2-enhances-stem-cell-fate-and-activates-expression-of-ascl2-in-intestinal-epithelial-cells
#16
Ashlee M Strubberg, Daniel A Veronese Paniagua, Tingting Zhao, Leeran Dublin, Thomas Pritchard, Peter O Bayguinov, James A J Fitzpatrick, Blair B Madison
Intestinal epithelial stem cell (IESC) fate is promoted by two major transcriptional regulators, the TCF4/β-catenin complex and ASCL2, which drive expression of IESC-specific factors, including Lgr5, Ephb2, and Rnf43. Canonical Wnt signaling via TCF4/β-catenin directly transactivates Ascl2, which in turn auto-regulates its own expression. Conversely, Let-7 microRNAs antagonize the IESC lineage by repressing specific mRNA targets. Here, we identify the zinc finger transcription factor PLAGL2 as a Let-7 target that regulates IESC fate...
July 9, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/30004386/slc6a14-an-amino-acid-transporter-modifies-the-primary-cf-defect-in-fluid-secretion
#17
Saumel Ahmadi, Sunny Xia, Yu-Sheng Wu, Michelle Di Paola, Randolph Kissoon, Catherine Luk, Fan Lin, Kai Du, Johanna Rommens, Christine E Bear
The severity of intestinal disease associated with Cystic Fibrosis (CF) is variable in the patient population and this variability is partially conferred by the influence of modifier genes. Genome-wide association studies have identified SLC6A14, an electrogenic amino acid transporter, as a genetic modifier of CF-associated meconium ileus. The purpose of the current work was to determine the biological role of Slc6a14, by disrupting its expression in CF mice bearing the major mutation, F508del. We found that disruption of Slc6a14 worsened the intestinal fluid secretion defect, characteristic of these mice...
July 13, 2018: ELife
https://www.readbyqxmd.com/read/29969439/optimized-base-editors-enable-efficient-editing-in-cells-organoids-and-mice
#18
Maria Paz Zafra, Emma M Schatoff, Alyna Katti, Miguel Foronda, Marco Breinig, Anabel Y Schweitzer, Amber Simon, Teng Han, Sukanya Goswami, Emma Montgomery, Jordana Thibado, Edward R Kastenhuber, Francisco J Sánchez-Rivera, Junwei Shi, Christopher R Vakoc, Scott W Lowe, Darjus F Tschaharganeh, Lukas E Dow
CRISPR base editing enables the creation of targeted single-base conversions without generating double-stranded breaks. However, the efficiency of current base editors is very low in many cell types. We reengineered the sequences of BE3, BE4Gam, and xBE3 by codon optimization and incorporation of additional nuclear-localization sequences. Our collection of optimized constitutive and inducible base-editing vector systems dramatically improves the efficiency by which single-nucleotide variants can be created...
July 3, 2018: Nature Biotechnology
https://www.readbyqxmd.com/read/29964038/vitamin-d-regulation-of-the-uridine-phosphorylase-1-gene-and-uridine-induced-dna-damage-in-colon-in-african-americans-and-european-americans
#19
Nobel Bhasin, Dereck Alleyne, Olivia A Gray, Sonia S Kupfer
BACKGROUND & AIMS: African Americans have the greatest colorectal cancer (CRC) burden in the United States-inter-ethnic differences in protective effects of vitamin D might contribute to disparities. 1α,25(OH)2 D3 vitamin D (the active form of vitamin D) induces transcription of the uridine phosphorylase gene (UPP1) in colon tissues from European Americans, but to a lesser extent in colon tissues from African Americans. UPP1-knockout mice have increased intestinal concentrations of uridine and dUTP, increased uridine-induced DNA damage, and develop colon tumors...
June 29, 2018: Gastroenterology
https://www.readbyqxmd.com/read/29959891/thrombin-modifies-growth-proliferation-and-apoptosis-of-human-colon-organoids-a-par1-and-par4-dependent-mechanism
#20
Morgane Sébert, Alexandre Denadai-Souza, Muriel Quaranta, Claire Racaud-Sultan, Sophie Chabot, Philippe Lluel, Nicolas Monjotin, Laurent Alric, Guillaume Portier, Sylvain Kirzin, Delphine Bonnet, Audrey Ferrand, Nathalie Vergnolle
BACKGROUND AND PURPOSE: Thrombin is massively released upon tissue damage associated with bleeding or chronic inflammation. The question of the effects of such thrombin on tissue regrowth and repair has been scarcely addressed, and only in cancer cell lines. The purpose of the present study was to determine thrombin's pharmacological effects on human intestinal epithelium growth, proliferation and apoptosis, using 3-dimentional cultures of human colon organoids. EXPERIMENTAL DESIGN: Crypts were isolated from human colonic resections and cultured for 6 days, forming human colon organoids...
June 30, 2018: British Journal of Pharmacology
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