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https://www.readbyqxmd.com/read/30534415/epigenetic-silencing-of-lgr5-induces-senescence-of-intestinal-epithelial-organoids-during-the-process-of-aging
#1
Ryoei Uchida, Yoshimasa Saito, Kazuki Nogami, Yohei Kajiyama, Yukana Suzuki, Yasuhiro Kawase, Toshiaki Nakaoka, Toshihide Muramatsu, Masaki Kimura, Hidetsugu Saito
To understand the molecular features underlying stem cell aging, we established intestinal epithelial organoids derived from both young and aged mice and investigated alterations in their senescence and epigenetic status. Senescence-related changes including accumulation of senescence-associated β-galactosidase and up-regulation of Cdkn1a ( p21) by DNA demethylation were observed in intestinal epithelial organoids derived from aged mice. We also demonstrated that the important stem cell marker Lgr5 was epigenetically silenced by trimethylation of histone H3 lysine 27, inducing suppression of Wnt signaling and a decrease of cell proliferation in organoids from aged mice...
2019: NPJ Aging and Mechanisms of Disease
https://www.readbyqxmd.com/read/30530633/mouse-fetal-intestinal-organoids-new-model-to-study-epithelial-maturation-from-suckling-to-weaning
#2
Marit Navis, Tânia Martins Garcia, Ingrid B Renes, Jacqueline Lm Vermeulen, Sander Meisner, Manon E Wildenberg, Gijs R van den Brink, Ruurd M van Elburg, Vanesa Muncan
During the suckling-to-weaning transition, the intestinal epithelium matures, allowing digestion of solid food. Transplantation experiments with rodent fetal epithelium into subcutaneous tissue of adult animals suggest that this transition is intrinsically programmed and occurs in the absence of dietary or hormonal signals. Here, we show that organoids derived from mouse primary fetal intestinal epithelial cells express markers of late fetal and neonatal development. In a stable culture medium, these fetal epithelium-derived organoids lose all markers of neonatal epithelium and start expressing hallmarks of adult epithelium in a time frame that mirrors epithelial maturation in vivo In vitro postnatal development of the fetal-derived organoids accelerates by dexamethasone, a drug used to accelerate intestinal maturation in vivo Together, our data show that organoids derived from fetal epithelium undergo suckling-to-weaning transition, that the speed of maturation can be modulated, and that fetal organoids can be used to model the molecular mechanisms of postnatal epithelial maturation...
December 10, 2018: EMBO Reports
https://www.readbyqxmd.com/read/30526881/human-intestinal-organoids-maintain-self-renewal-capacity-and-cellular-diversity-in-niche-inspired-culture-condition
#3
Masayuki Fujii, Mami Matano, Kohta Toshimitsu, Ai Takano, Yohei Mikami, Shingo Nishikori, Shinya Sugimoto, Toshiro Sato
Cellular diversity that shapes tissue architecture and function is governed by multiple niche signals. Nonetheless, maintaining cellular diversity in human intestinal organoids has been challenging. Based on niche ligands present in the natural stem cell milieu, we establish a refined organoid culture condition for intestinal epithelia that allows human intestinal organoids to concurrently undergo multi-differentiation and self-renewal. High-throughput screening reveals that the combination of insulin-like growth factor 1 (IGF-1) and fibroblast growth factor 2 (FGF-2) enhances the clonogenic capacity and CRISPR-genome engineering efficiency of human intestinal stem cells...
December 6, 2018: Cell Stem Cell
https://www.readbyqxmd.com/read/30521806/small-heterodimer-partner-and-fibroblast-growth-factor-19-inhibit-expression-of-npc1l1-in-mouse-intestine-and-cholesterol-absorption
#4
Young-Chae Kim, Sangwon Byun, Sunmi Seok, Grace Guo, H Eric Xu, Byron Kemper, Jongsook Kim Kemper
BACKGROUND & AIMS: The nuclear receptor subfamily 0 group B member 2 (NR0B2, also called SHP) is expressed at high levels in liver and intestine. Postprandial fibroblast growth factor 19 (human FGF19, mouse FGF15) signaling increases the transcriptional activity of SHP. We studied the functions of SHP and FGF19 in intestines of mice, including their regulation of expression of the cholesterol transporter NPC1-like intracellular cholesterol transporter 1 (NPC1L1) and cholesterol absorption...
December 3, 2018: Gastroenterology
https://www.readbyqxmd.com/read/30504008/high-throughput-single-cell-transcriptomics-on-organoids
#5
REVIEW
Agnieska Brazovskaja, Barbara Treutlein, J Gray Camp
Three-dimensional (3D) tissues grown in culture from human stem cells offer the incredible opportunity to analyze and manipulate human development, and to generate patient-specific models of disease. Methods to sequence DNA and RNA in single cells are being used to analyze these so-called 'organoid' systems in high-resolution. Single-cell transcriptomics has been used to quantitate the similarity of organoid cells to primary tissue counterparts in the brain, intestine, liver, and kidney, as well as identify cell-specific responses to environmental variables and disease conditions...
November 29, 2018: Current Opinion in Biotechnology
https://www.readbyqxmd.com/read/30503340/leucine-rich-repeat-containing-g-protein-coupled-receptor-5-enriched-organoids-under-chemically-defined-growth-conditions
#6
Soojung Hahn, Mi Sun Kim, Seon Young Choi, Sukin Jeong, JooHyun Jee, Han Kyung Kim, Sang Yun Jeong, Hyunsoo Shin, Hyoung Sun Kim, Joon Seong Park, Jongman Yoo
An organoid is a complex, multi-cell three-dimensional (3D) structure that contains tissue-specific cells. Epithelial stem cells, which are marked by leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5), have the potential for self-renewal and expansion as organoids. However, in the case of intestinal organoids from Lgr5-EGFP-IRES-CreERT2 transgenic mice, in vitro expansion of the Lgr5 expression is limited in a culture condition supplemented with essential proteins, such as epidermal growth factor (E), noggin (N), and R-spondin 1 (R)...
November 28, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/30488522/the-effect-of-ptc-mutations-on-cftr-mrna-abundance-in-human-nasal-epithelium-and-intestinal-organoids-a-basis-for-read-through-therapies-in-cystic-fibrosis
#7
Luka A Clarke, Nikhil T Awatade, Veronica M Felicio, Iris A Silva, Maite Calucho, Luisa Pereira, Pilar Azevedo, José Cavaco, Celeste Barreto, Carmen Bertuzzo, Silvia Gartner, Jeffrey Beekman, Margarida D Amaral
A major challenge in Cystic Fibrosis research is applying mutation-specific therapy to individual patients with diverse and rare CFTR genotypes. Read-through agents are currently the most promising approach for Class I mutations that introduce premature termination codons (PTCs) into CFTR mRNA. However, variations in degradation of PTC containing transcripts by nonsense mediated decay (NMD) might lower read-through efficacy. Allele specific quantitative real time (qRT-)PCR was used to measure variations in CFTR mRNA abundance for several PTC mutations in respiratory cells and intestinal organoids...
November 28, 2018: Human Mutation
https://www.readbyqxmd.com/read/30478383/dclk1-in-tuft-cells-promotes-inflammation-driven-epithelial-restitution-and-mitigates-chronic-colitis
#8
Jun Yi, Kirk Bergstrom, Jianxin Fu, Xindi Shan, J Michael McDaniel, Samuel McGee, Dongfeng Qu, Courtney W Houchen, Xiaowei Liu, Lijun Xia
Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by defective intestinal barrier integrity toward the microbiota and epithelial damage. Double cortin-like kinase 1 (Dclk1), a marker of intestinal tuft cells, can regulate tissue regenerative responses, but its role in epithelial repair during bacterial-dependent chronic colitis is unclear. We addressed this question using our recently developed mouse model of spontaneous microbiota-dependent colitis induced by mucin-type O-glycan deficiency (DKO), which recapitulates most features of human UC...
November 26, 2018: Cell Death and Differentiation
https://www.readbyqxmd.com/read/30478287/an-upstream-protein-coding-region-in-enteroviruses-modulates-virus-infection-in-gut-epithelial-cells
#9
Valeria Lulla, Adam M Dinan, Myra Hosmillo, Yasmin Chaudhry, Lee Sherry, Nerea Irigoyen, Komal M Nayak, Nicola J Stonehouse, Matthias Zilbauer, Ian Goodfellow, Andrew E Firth
Enteroviruses comprise a large group of mammalian pathogens that includes poliovirus. Pathology in humans ranges from sub-clinical to acute flaccid paralysis, myocarditis and meningitis. Until now, all of the enteroviral proteins were thought to derive from the proteolytic processing of a polyprotein encoded in a single open reading frame. Here we report that many enterovirus genomes also harbour an upstream open reading frame (uORF) that is subject to strong purifying selection. Using echovirus 7 and poliovirus 1, we confirmed the expression of uORF protein in infected cells...
November 26, 2018: Nature Microbiology
https://www.readbyqxmd.com/read/30469504/2d-and-3d-based-intestinal-stem-cell-cultures-for-personalized-medicine
#10
REVIEW
Yuan Liu, Ye-Guang Chen
Colorectal cancer (CRC) is one of the most common cancers that have high occurrence and death in both males and females. As various factors have been found to contribute to CRC development, personalized therapies are critical for efficient treatment. To achieve this purpose, the establishment of patient-derived tumor models is critical for diagnosis and drug test. The establishment of three-dimensional (3D) organoid cultures and two-dimensional (2D) monolayer cultures of patient-derived epithelial tissues is a breakthrough for expanding living materials for later use...
November 22, 2018: Cells
https://www.readbyqxmd.com/read/30465020/nhe8-deficiency-promotes-colitis-associated-cancer-in-mice-via-expansion-of-lgr5-expressing-cells
#11
Hua Xu, Jing Li, Hao Chen, Fayez K Ghishan
Background & Aims: Lgr5 overexpression has been detected in colorectal cancers (CRCs), including some cases of colitis-associated CRCs. In colitis-associated CRCs, chronic inflammation is a contributing factor in carcinogenesis. We recently reported that intestinal Na+ /H+ exchanger isoform 8 (NHE8) plays an important role in intestinal mucosal protection and that loss of NHE8 expression results in an ulcerative colitis-like condition. Therefore, we hypothesized that NHE8 may be involved in the development of intestinal tumors...
2019: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/30455690/atf3-sustains-il-22-induced-stat3-phosphorylation-to-maintain-mucosal-immunity-through-inhibiting-phosphatases
#12
Doaa Glal, Janaki N Sudhakar, Hsueh-Han Lu, Ming-Che Liu, Hung-Yu Chiang, Yen-Chun Liu, Ching-Feng Cheng, Jr-Wen Shui
In gut epithelium, IL-22 transmits signals through STAT3 phosphorylation (pSTAT3) which provides intestinal immunity. Many components in the IL-22-pSTAT3 pathway have been identified as risk factors for inflammatory bowel disease (IBD) and some of them are considered as promising therapeutic targets. However, new perspectives are still needed to understand IL-22-pSTAT3 signaling for effective clinical interventions in IBD patients. Here, we revealed activating transcription factor 3 (ATF3), recently identified to be upregulated in patients with active IBD, as a crucial player in the epithelial IL-22-pSTAT3 signaling cascade...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/30449703/genome-toxicity-and-impaired-stem-cell-function-after-conditional-activation-of-creer-t2-in-the-intestine
#13
Natacha Bohin, Elizabeth A Carlson, Linda C Samuelson
With the tamoxifen-inducible CreERT2 system, genetic recombination can be temporally controlled in a cell-type-specific manner in intact animals, permitting dissection of the molecular underpinnings of mammalian physiology. Here we present a significant drawback to CreERT2 technology for analysis of intestinal stem cells. Using the intestine-specific Villin-CreERT2 mouse strain, we observed delayed intestinal regeneration post irradiation. Villin-CreERT2 activation was associated with DNA damage and cryptic loxP site cleavage...
October 31, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/30443904/organ-on-chip-models-implications-in-drug-discovery-and-clinical-applications
#14
REVIEW
Rahul Mittal, Frank W Woo, Carlo S Castro, Madeline A Cohen, Joana Karanxha, Jeenu Mittal, Tanya Chhibber, Vasanti M Jhaveri
Before a lead compound goes through a clinical trial, preclinical studies utilize two-dimensional (2D) in vitro models and animal models to study the pharmacodynamics and pharmacokinetics of that lead compound. However, these current preclinical studies may not accurately represent the efficacy and safety of a lead compound in humans, as there has been a high failure rate of drugs that enter clinical trials. All of these failures and the associated costs demonstrate a need for more representative models of human organ systems for screening in the preclinical phase of drug development...
November 15, 2018: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/30430443/correction-to-the-three-dimensional-culture-of-epithelial-organoids-derived-from-embryonic-chicken-intestine
#15
Malgorzata Pierzchalska, Malgorzata Panek, Malgorzata Czyrnek, Maja Grabacka
Erratum to: The Three-Dimensional Culture of Epithelial Organoids Derived from Embryonic Chicken Intestine.
November 11, 2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/30418409/bacteriophage-therapy-testing-against-shigella-flexneri-in-a-novel-human-intestinal-organoid-derived-infection-model
#16
Alejandro Llanos-Chea, Robert J Citorik, Kourtney P Nickerson, Laura Ingano, Gloria Serena, Stefania Senger, Timothy K Lu, Alessio Fasano, Christina S Faherty
OBJECTIVE: Enteric bacterial pathogens cause diarrheal disease and mortality at significant rates throughout the world, particularly in children under the age of 5 years. Our ability to combat bacterial pathogens has been hindered antibiotic resistance, a lack of effective vaccines, and accurate models of infection. With the renewed interest in bacteriophage therapy, we sought to use a novel human intestinal model to investigate the efficacy of a newly isolated bacteriophage against Shigella flexneri...
November 8, 2018: Journal of Pediatric Gastroenterology and Nutrition
https://www.readbyqxmd.com/read/30409423/a-novel-morphometry-system-automatically-assessing-the-growth-and-regeneration-of-intestinal-organoids
#17
Jiong Ren, Zhibin Niu, Xiaoqin Li, Jie Yang, Meijiao Gao, Xudong Li, Tao Zhang, Lei Fang, Boyang Zhang, Junping Wang, Yongping Su, Fengchao Wang
As compared with 2D cell line cultures, 3D intestinal organoids are better at maximally recapitulating the physiological features of stem cells in vivo. However, the complex 3D structure is an obstacle which must be objectively and automatically evaluated to assess colony growth and regeneration. Meanwhile, no internal standard currently exists for evaluating the size of heterogeneities in organoids or defining those regenerating colonies. Herein, we developed a simple morphometry system to image MTT-stained organoids...
November 5, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/30407734/human-milk-oligosaccharides-increase-mucin-expression-in-experimental-necrotizing-enterocolitis
#18
Richard You Wu, Bo Li, Yuhki Koike, Pekka Määttänen, Hiromu Miyake, Marissa Cadete, Kathene C Johnson-Henry, Steven R Botts, Carol Lee, Thomas R Abrahamsson, Eva Landberg, Agostino Pierro, Philip M Sherman
SCOPE: Necrotizing enterocolitis (NEC) is a leading cause of morbidity and death in preterm infants, occurring more often in formula-fed than breastfed infants. Studies in both rats and humans show that human milk oligosaccharides (HMOs) lower the incidence of NEC, but the mechanism underlying such protection is currently unclear. METHODS AND RESULTS: Here by extracting HMOs from pooled human breastmilk, we investigated the impact of HMOs on the intestinal mucin levels in a murine model of NEC...
November 8, 2018: Molecular Nutrition & Food Research
https://www.readbyqxmd.com/read/30389919/arts-mediates-apoptosis-and-regeneration-of-the-intestinal-stem-cell-niche
#19
Elle Koren, Yahav Yosefzon, Roi Ankawa, Despina Soteriou, Avi Jacob, Alexander Nevelsky, Rahamim Ben-Yosef, Gil Bar-Sela, Yaron Fuchs
Stem cells (SCs) play a pivotal role in fueling homeostasis and regeneration. While much focus has been given to self-renewal and differentiation pathways regulating SC fate, little is known regarding the specific mechanisms utilized for their elimination. Here, we report that the pro-apoptotic protein ARTS (a Septin4 isoform) is highly expressed in cells comprising the intestinal SC niche and that its deletion protects Lgr5+ and Paneth cells from undergoing apoptotic cell death. As a result, the Sept4/ARTS-/- crypt displays augmented proliferation and, in culture, generates massive cystic-like organoids due to enhanced Wnt/β-catenin signaling...
November 2, 2018: Nature Communications
https://www.readbyqxmd.com/read/30382376/neonatal-intestinal-organoids-as-an-ex-vivo-approach-to-study-early-intestinal-epithelial-disorders
#20
Bo Li, Carol Lee, Marissa Cadete, Hiromu Miyake, Dorothy Lee, Agostino Pierro
BACKGROUND: Adult intestinal organoids have been used to study ex vivo intestinal injury in adulthood. However, the neonatal intestinal epithelium has many unique features that are different from adult mature intestine. Establishing a neonatal ex vivo organoid model is essential to study the epithelial physiology in early postnatal development and to investigate derangements associated with disease processes during the neonatal period like necrotizing enterocolitis (NEC). METHODS: Fresh and frozen terminal ileum was harvested from mice pups on postnatal day 9...
October 31, 2018: Pediatric Surgery International
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