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https://www.readbyqxmd.com/read/29656244/human-organoid-cultures-transformative-new-tools-for-human-virus-studies
#1
REVIEW
Sasirekha Ramani, Sue E Crawford, Sarah E Blutt, Mary K Estes
Studies of human infectious diseases have been limited by the paucity of functional models that mimic normal human physiology and pathophysiology. Recent advances in the development of multicellular, physiologically active organotypic cultures produced from embryonic and pluripotent stem cells, as well as from stem cells isolated from biopsies and surgical specimens are allowing unprecedented new studies and discoveries about host-microbe interactions. Here, we summarize recent developments in the use of organoids for studying human viral pathogens, including intestinal infections with human rotavirus, norovirus, enteroviruses and adenoviruses (intestinal organoids and enteroids), neuronal infections with Zika virus (cerebral organoids) and respiratory infections with respiratory syncytial virus in (lung bud organoids)...
April 12, 2018: Current Opinion in Virology
https://www.readbyqxmd.com/read/29649441/small-intestine-microbiota-regulate-host-digestive-and-absorptive-adaptive-responses-to-dietary-lipids
#2
Kristina Martinez-Guryn, Nathaniel Hubert, Katya Frazier, Saskia Urlass, Mark W Musch, Patricia Ojeda, Joseph F Pierre, Jun Miyoshi, Timothy J Sontag, Candace M Cham, Catherine A Reardon, Vanessa Leone, Eugene B Chang
The gut microbiota play important roles in lipid metabolism and absorption. However, the contribution of the small bowel microbiota of mammals to these diet-microbe interactions remains unclear. We determine that germ-free (GF) mice are resistant to diet-induced obesity and malabsorb fat with specifically impaired lipid digestion and absorption within the small intestine. Small bowel microbes are essential for host adaptation to dietary lipid changes by regulating gut epithelial processes involved in their digestion and absorption...
April 11, 2018: Cell Host & Microbe
https://www.readbyqxmd.com/read/29643510/intra-tumour-diversification-in-colorectal-cancer-at-the-single-cell-level
#3
Sophie F Roerink, Nobuo Sasaki, Henry Lee-Six, Matthew D Young, Ludmil B Alexandrov, Sam Behjati, Thomas J Mitchell, Sebastian Grossmann, Howard Lightfoot, David A Egan, Apollo Pronk, Niels Smakman, Joost van Gorp, Elizabeth Anderson, Stephen J Gamble, Chris Alder, Marc van de Wetering, Peter J Campbell, Michael R Stratton, Hans Clevers
Every cancer originates from a single cell. During expansion of the neoplastic cell population, individual cells acquire genetic and phenotypic differences from each other. Here, to investigate the nature and extent of intra-tumour diversification, we characterized organoids derived from multiple single cells from three colorectal cancers as well as from adjacent normal intestinal crypts. Colorectal cancer cells showed extensive mutational diversification and carried several times more somatic mutations than normal colorectal cells...
April 11, 2018: Nature
https://www.readbyqxmd.com/read/29643147/-ex-vivo-gut-culture-for-studying-differentiation-and-migration-of-small-intestinal-epithelial-cells
#4
Xiaofei Sun, Xing Fu, Min Du, Mei-Jun Zhu
Epithelial cultures are commonly used for studying gut health. However, due to the absence of mesenchymal cells and gut structure, epithelial culture systems including recently developed three-dimensional organoid culture cannot accurately represent in vivo gut development, which requires intense cross-regulation of the epithelial layer with the underlying mesenchymal tissue. In addition, organoid culture is costly. To overcome this, a new culture system was developed using mouse embryonic small intestine. Cultured intestine showed spontaneous peristalsis, indicating the maintenance of the normal gut physiological structure...
April 2018: Open Biology
https://www.readbyqxmd.com/read/29621481/in-inflamed-intestinal-tissues-and-epithelial-cells-interleukin-22-signaling-increases-expression-of-h19-long-noncoding-rna-which-promotes-mucosal-regeneration
#5
Hua Geng, Heng-Fu Bu, Fangyi Liu, Longtao Wu, Karl Pfeifer, Pauline M Chou, Xiao Wang, Jiaren Sun, Lu Lu, Ashutosh Pandey, Marisa S Bartolomei, Isabelle G De Plaen, Peng Wang, Jindan Yu, Jiaming Qian, Xiao-Di Tan
BACKGROUND & AIMS: Inflammation affects regeneration of the intestinal epithelia; long non-coding RNAs (lncRNAs) regulate cell functions such as proliferation, differentiation, and migration. We investigated the mechanisms by which the lncRNA H19, imprinted maternally expressed transcript (H19) regulates regeneration of intestinal epithelium using cell cultures and mouse models of inflammation. METHODS: We performed RNA-seq transcriptome analyses of intestinal tissues from mice with LPS-induced sepsis to identify lncRNAs associated with inflammation; findings were confirmed by quantitative real-time PCR and in situ hybridization analyses of intestinal tissues from mice with sepsis or dextran sulfate sodium (DSS)-induced mucosal wound healing and patients with ulcerative colitis, compared to healthy individuals (controls)...
April 2, 2018: Gastroenterology
https://www.readbyqxmd.com/read/29615438/generation-of-intestinal-organoids-suitable-for-pharmacokinetic-studies-from-human-induced-pluripotent-stem-cells
#6
Daichi Onozato, Misaki Yamashita, Anna Nakanishi, Takumi Akagawa, Yuriko Kida, Isamu Ogawa, Tadahiro Hashita, Takahiro Iwao, Tamihide Matsunaga
Intestinal organoids morphologically resemble intestinal tissues, and are expected to be used in both regenerative medicine and drug development studies, including pharmacokinetic studies. However, the pharmacokinetic properties of these organoids remain poorly characterized. In this study, we aimed to generate pharmacokinetically functional intestinal organoids from human induced pluripotent stem (iPS) cells. Human iPS cells were induced to differentiate into the hindgut and then seeded on EZSPHERE plates to generate uniform spheroids, and the floating spheroids were subsequently differentiated into intestinal organoids by using small-molecule compounds...
April 3, 2018: Drug Metabolism and Disposition: the Biological Fate of Chemicals
https://www.readbyqxmd.com/read/29604290/intestinal-failure-and-aberrant-lipid-metabolism-in-patients-with-dgat1-deficiency
#7
Jorik M van Rijn, Rico Chandra Ardy, Zarife Kuloğlu, Bettina Härter, Désirée Y van Haaften-Visser, Hubert P J van der Doef, Marliek van Hoesel, Aydan Kansu, Anke H M van Vugt, Marini Ng, Freddy T M Kokke, Ana Krolo, Meryem Keçeli Başaran, Neslihan Gurcan Kaya, Aysel Ünlüsoy Aksu, Buket Dalgıç, Figen Ozcay, Zeren Baris, Renate Kain, Edwin C A Stigter, Klaske D Lichtenbelt, Maarten P G Massink, Karen J Duran, Joke B G M Verheij, Dorien Lugtenberg, Peter G J Nikkels, Henricus G F Brouwer, Henkjan J Verkade, Rene Scheenstra, Bart Spee, Edward E S Nieuwenhuis, Paul J Coffer, Andreas R Janecke, Gijs van Haaften, Roderick H J Houwen, Thomas Müller, Sabine Middendorp, Kaan Boztug
BACKGROUND & AIMS: Congenital diarrheal disorders are rare inherited intestinal disorders characterized by intractable, sometimes life-threatening, diarrhea and nutrient malabsorption; some have been associated with mutations in diacylglycerol-acyltransferase 1 (DGAT1), which catalyzes formation of triacylglycerol from diacylglycerol and acyl-CoA. We investigated the mechanisms by which DGAT1 deficiency contributes to intestinal failure using patient-derived organoids. METHODS: We collected blood samples from 10 patients, from 6 unrelated pedigrees, who presented with early-onset severe diarrhea and/or vomiting, hypoalbuminemia, and/or (fatal) protein-losing enteropathy with intestinal failure; we performed next-generation sequence analysis of DNA from 8 patients...
March 28, 2018: Gastroenterology
https://www.readbyqxmd.com/read/29596476/the-colonic-epithelium-plays-an-active-role-in-promoting-colitis-by-shaping-the-tissue-cytokine-profile
#8
Jesse Lyons, Phaedra C Ghazi, Alina Starchenko, Alessio Tovaglieri, Katherine R Baldwin, Emily J Poulin, Jessica J Gierut, Casie Genetti, Vijay Yajnik, David T Breault, Douglas A Lauffenburger, Kevin M Haigis
Inflammatory bowel disease (IBD) is a chronic condition driven by loss of homeostasis between the mucosal immune system, the commensal gut microbiota, and the intestinal epithelium. Our goal is to understand how these components of the intestinal ecosystem cooperate to control homeostasis. By combining quantitative measures of epithelial hyperplasia and immune infiltration with multivariate analysis of inter- and intracellular signaling, we identified epithelial mammalian target of rapamycin (mTOR) signaling as a potential driver of inflammation in a mouse model of colitis...
March 2018: PLoS Biology
https://www.readbyqxmd.com/read/29590640/intestinal-epithelial-cell-polarity-defects-in-disease-lessons-from-microvillus-inclusion-disease
#9
REVIEW
Kerstin Schneeberger, Sabrina Roth, Edward E S Nieuwenhuis, Sabine Middendorp
The intestinal epithelium is a highly organized tissue. The establishment of epithelial cell polarity, with distinct apical and basolateral plasma membrane domains, is pivotal for both barrier formation and for the uptake and vectorial transport of nutrients. The establishment of cell polarity requires a specialized subcellular machinery to transport and recycle proteins to their appropriate location. In order to understand and treat polarity-associated diseases, it is necessary to understand epithelial cell-specific trafficking mechanisms...
February 13, 2018: Disease Models & Mechanisms
https://www.readbyqxmd.com/read/29589318/use-of-organoids-to-characterize-signaling-pathways-in-cancer-initiation
#10
Christina Oatway, Calley L Hirsch, Alex Gregorieff
The development of intestinal organoid technology has greatly accelerated research in the field of colorectal cancer. Contrary to traditional cancer cell lines, organoids are composed of multiple cell types arranged in 3D structures highly reminiscent of their native tissues. Thus, organoids provide a near-physiological and readily accessible model to study tissue morphogenesis, adult stem cell behavior and tumorigenesis. Here, we provide protocols for establishing intestinal organoid cultures from genetically modified mouse lines and describe methods to overexpress and knockout genes of interest using lentiviral-based approaches...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29589266/efficient-culture-of-intestinal-organoids-with-blebbistatin
#11
Zhen Qi, Ye-Guang Chen
The intestinal epithelium is one of the most rapidly self-renewing tissues throughout life in mammals. A small population of stem cells at the base of crypt in the epithelium can continually self-renew and give rise to differentiated epithelial cells. The self-renewal and differentiation of intestinal stem cells are under a tight control during homeostasis, and disruption of this balancing regulation leads to intestinal degeneration or tumorigenesis. Accordingly, exploration of the mechanism underlying the regulation of stem cells is essential for the understanding and treatment of intestinal disorders...
September 27, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29579172/micrornas-in-the-mammalian-gut-endocrine-lineage
#12
Yu-Han Hung, Praveen Sethupathy
MicroRNAs (miRNAs) are small noncoding RNA molecules that modulate gene expression at the posttranscriptional level. Numerous reports have elucidated the importance of miRNAs in the regulation of a wide array of biological processes including metabolism and energy homeostasis. miRNAs in the endocrine pancreas have been intensively studied over the last 15 years and linked to pancreatic islet development and function. In comparison, knowledge of miRNAs in gut endocrine cells, or enteroendocrine cells (EECs), is severely lacking...
February 1, 2018: Endocrinology
https://www.readbyqxmd.com/read/29562732/indole-3-carbinol-promotes-goblet-cell-differentiation-regulating-wnt-and-notch-signaling-pathways-ahr-dependently
#13
Joo-Hung Park, Jeong-Min Lee, Eun-Jin Lee, Won-Bhin Hwang, Da-Jeong Kim
Using an in vitro model of intestinal organoids derived from intestinal crypts, we examined effects of indole-3-carbinol (I3C), a phytochemical that has anticancer and aryl hydrocarbon receptor (AhR)-activating abilities and thus is sold as a dietary supplement, on the development of intestinal organoids and investigated the underlying mechanisms. I3C inhibited the in vitro development of mouse intestinal organoids. Addition of α-naphthoflavone, an AhR antagonist or AhR siRNA transfection, suppressed I3C function, suggesting that I3C-mediated interference with organoid development is AhR-dependent...
March 21, 2018: Molecules and Cells
https://www.readbyqxmd.com/read/29559533/-pdgfr%C3%AE-pericryptal-stromal-cells-are-the-critical-source-of-wnts-and-rspo3-for-murine-intestinal-stem-cells-in-vivo
#14
Gediminas Greicius, Zahra Kabiri, Kristmundur Sigmundsson, Chao Liang, Ralph Bunte, Manvendra K Singh, David M Virshup
Wnts and R-spondins (RSPOs) support intestinal homeostasis by regulating crypt cell proliferation and differentiation. Ex vivo, Wnts secreted by Paneth cells in organoids can regulate the proliferation and differentiation of Lgr5 -expressing intestinal stem cells. However, in vivo, Paneth cell and indeed all epithelial Wnt production is completely dispensable, and the cellular source of Wnts and RSPOs that maintain the intestinal stem-cell niche is not known. Here we investigated both the source and the functional role of stromal Wnts and RSPO3 in regulation of intestinal homeostasis...
March 20, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29552623/translating-developmental-principles-to-generate-human-gastric-organoids
#15
REVIEW
Alexandra K Eicher, H Matthew Berns, James M Wells
Gastric diseases, including peptic ulcer disease and gastric cancer, are highly prevalent in human beings. Despite this, the cellular biology of the stomach remains poorly understood relative to other gastrointestinal organs such as the liver, intestine, and colon. In particular, little is known about the molecular basis of stomach development and the differentiation of gastric lineages. Although animal models are useful for studying gastric development, function, and disease, there are major structural and physiological differences in human stomachs that render these models insufficient...
March 2018: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/29544685/comparison-of-ex-vivo-and-in-vitro-intestinal-cystic-fibrosis-models-to-measure-cftr-dependent-ion-channel-activity
#16
Domenique D Zomer-van Ommen, Eyleen de Poel, Evelien Kruisselbrink, Hugo Oppelaar, Annelotte M Vonk, Hettie M Janssens, Cornelis K van der Ent, Marne C Hagemeijer, Jeffrey M Beekman
BACKGROUND: New functional assays using primary human intestinal adult stem cell cultures can be valuable tools to study epithelial defects in human diseases such as cystic fibrosis. METHODS: CFTR-mediated ion transport was measured in rectal organoid-derived monolayers grown from subjects with various CFTR mutations and compared to donor-matched intestinal current measurements (ICM) in rectal biopsies and forskolin-induced swelling of rectal organoids. RESULTS: Rectal organoid-derived monolayers were generated within four days...
March 13, 2018: Journal of Cystic Fibrosis: Official Journal of the European Cystic Fibrosis Society
https://www.readbyqxmd.com/read/29538044/mechanisms-for-intestinal-regeneration
#17
Genia Dubrovsky, James C Y Dunn
PURPOSE OF REVIEW: The purpose of this review is to briefly summarize the notable structures and pathways in intestinal epithelial growth before presenting the current main areas of active research in intestinal regeneration. As a rapidly advancing field, a number of breakthroughs have recently been made related to the culture of intestinal stem cells (ISCs) and to the engineering of intestinal tissue. RECENT FINDINGS: ISCs can be derived from fibroblasts and can be cultured in hydrogels under xenogeneic-free conditions...
March 13, 2018: Current Opinion in Pediatrics
https://www.readbyqxmd.com/read/29535828/morphological-alterations-of-cultured-human-colorectal-matched-tumour-and-healthy-organoids
#18
Seyed Mohammad Hossein Kashfi, Sheema Almozyan, Nicholas Jinks, Bon-Kyoung Koo, Abdolrahman S Nateri
Organoids have extensive applications in many fields ranging from modelling human development and disease, personalised medicine, drug screening, etc. Moreover, in the last few years, several studies have evaluated the capacity of organoids as transplantation sources for therapeutic approaches and regenerative medicine. Nevertheless, depending on the origin of the cells and anatomical complications, an organoid transplant may make tissue regeneration difficult. However, some essential aspects of organoids including the morphological alterations and the growth pattern of the matched tumour and their healthy derived organoids have received less attention...
February 13, 2018: Oncotarget
https://www.readbyqxmd.com/read/29535731/nlrx1-modulates-immunometabolic-mechanisms-controlling-the-host-gut-microbiota-interactions-during-inflammatory-bowel-disease
#19
Andrew Leber, Raquel Hontecillas, Nuria Tubau-Juni, Victoria Zoccoli-Rodriguez, Vida Abedi, Josep Bassaganya-Riera
Interactions among the gut microbiome, dysregulated immune responses, and genetic factors contribute to the pathogenesis of inflammatory bowel disease (IBD). Nlrx1 -/- mice have exacerbated disease severity, colonic lesions, and increased inflammatory markers. Global transcriptomic analyses demonstrate enhanced mucosal antimicrobial defense response, chemokine and cytokine expression, and epithelial cell metabolism in colitic Nlrx1 -/- mice compared to wild-type (WT) mice. Cell-specificity studies using cre-lox mice demonstrate that the loss of NLRX1 in intestinal epithelial cells (IEC) recapitulate the increased sensitivity to DSS colitis observed in whole body Nlrx1 -/- mice...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29534451/stem-cell-derived-models-of-viral-infections-in-the-gastrointestinal-tract
#20
REVIEW
Wyatt E Lanik, Madison A Mara, Belgacem Mihi, Carolyn B Coyne, Misty Good
Studies on the intestinal epithelial response to viral infection have previously been limited by the absence of in vitro human intestinal models that recapitulate the multicellular complexity of the gastrointestinal tract. Recent technological advances have led to the development of "mini-intestine" models, which mimic the diverse cellular nature and physiological activity of the small intestine. Utilizing adult or embryonic intestinal tissue, enteroid and organoid systems, respectively, represent an opportunity to effectively model cellular differentiation, proliferation, and interactions that are specific to the specialized environment of the intestine...
March 10, 2018: Viruses
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