67 results on '"Stappenbeck, Thaddeus S."'
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2. Su1968 CAUSES IN CLINICAL TRIALS FAILURE OF INFLAMMATORY BOWEL DISEASE (IBD): AN ARTIFICIAL INTELLIGENCE- ASSISTED REVIEW.
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Yadete, Tesfaye, Njoku, Kingsley, Bettencourt-Silva, Joao, Liu, Matthew, Anand, Vibha, Kartoun, Uri, Mulligan, Natasha, Koski, Eileen, Stappenbeck, Thaddeus S., and Liu, Julia J.
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- 2024
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3. 981 WESTERN DIET DEPLETES SMALL INTESTINAL INTRAEPITHELIAL LYMPHOCYTES VIA FXR-TYPE I INTERFERON PATHWAY.
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Ma, Changqing, Trsan, Tihana, Mishra, Richa, Frein, Jennifer, Kern, Justin, Jain, Umang, Colonna, Marco, Stappenbeck, Thaddeus S., and Liu, Ta-Chiang
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- 2024
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4. HOIL1 regulates group 2 innate lymphoid cell numbers and type 2 inflammation in the small intestine
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Wood, Matthew J., Marshall, Jeffrey N., Hartley, Victoria L., Liu, Ta-Chiang, Iwai, Kazuhiro, Stappenbeck, Thaddeus S., and MacDuff, Donna A.
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Patients with mutations in HOIL1experience a complex immune disorder including intestinal inflammation. To investigate the role of HOIL1in regulating intestinal inflammation, we employed a mouse model of partial HOIL1 deficiency. The ileum of HOIL1-deficient mice displayed features of type 2 inflammation including tuft cell and goblet cell hyperplasia, and elevated expression of Il13, Il5and Il25mRNA. Inflammation persisted in the absence of T and B cells, and bone marrow chimeric mice revealed a requirement for HOIL1 expression in radiation-resistant cells to regulate inflammation. Although disruption of IL-4 receptor alpha (IL4Rα) signaling on intestinal epithelial cells ameliorated tuft and goblet cell hyperplasia, expression of Il5and Il13mRNA remained elevated. KLRG1hiCD90logroup 2 innate lymphoid cells were increased independent of IL4Rα signaling, tuft cell hyperplasia and IL-25 induction. Antibiotic treatment dampened intestinal inflammation indicating commensal microbes as a contributing factor. We have identified a key role for HOIL1, a component of the Linear Ubiquitin Chain Assembly Complex, in regulating type 2 inflammation in the small intestine. Understanding the mechanism by which HOIL1 regulates type 2 inflammation will advance our understanding of intestinal homeostasis and inflammatory disorders and may lead to the identification of new targets for treatment.
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- 2022
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5. Organoids in gastrointestinal diseases: from experimental models to clinical translation
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Gu¨nther, Claudia, Winner, Beate, Neurath, Markus F, and Stappenbeck, Thaddeus S
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We are entering an era of medicine where increasingly sophisticated data will be obtained from patients to determine proper diagnosis, predict outcomes and direct therapies. We predict that the most valuable data will be produced by systems that are highly dynamic in both time and space. Three-dimensional (3D) organoids are poised to be such a highly valuable system for a variety of gastrointestinal (GI) diseases. In the lab, organoids have emerged as powerful systems to model molecular and cellular processes orchestrating natural and pathophysiological human tissue formation in remarkable detail. Preclinical studies have impressively demonstrated that these organs-in-a-dish can be used to model immunological, neoplastic, metabolic or infectious GI disorders by taking advantage of patient-derived material. Technological breakthroughs now allow to study cellular communication and molecular mechanisms of interorgan cross-talk in health and disease including communication along for example, the gut–brain axis or gut–liver axis. Despite considerable success in culturing classical 3D organoids from various parts of the GI tract, some challenges remain to develop these systems to best help patients. Novel platforms such as organ-on-a-chip, engineered biomimetic systems including engineered organoids, micromanufacturing, bioprinting and enhanced rigour and reproducibility will open improved avenues for tissue engineering, as well as regenerative and personalised medicine. This review will highlight some of the established methods and also some exciting novel perspectives on organoids in the fields of gastroenterology. At present, this field is poised to move forward and impact many currently intractable GI diseases in the form of novel diagnostics and therapeutics.
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- 2022
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6. Reverse translation approach generates a signature of penetrating fibrosis in Crohn’s disease that is associated with anti-TNF response
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Xiong, Shanshan, Whitehurst, Charles E, Li, Li, Heo, Gyu Seong, Lai, Chin-Wen, Jain, Umang, Muegge, Brian D, Espenschied, Scott T, Musich, Ryan J, Chen, Minhu, Liu, Yongjian, Liu, Ta-Chiang, and Stappenbeck, Thaddeus S
- Abstract
ObjectiveFibrosis is a common feature of Crohn’s disease (CD) which can involve the mesenteric fat. However, the molecular signature of this process remains unclear. Our goal was to define the transcriptional signature of mesenteric fibrosis in CD subjects and to model mesenteric fibrosis in mice to improve our understanding of CD pathogenesis.DesignWe performed histological and transcriptional analysis of fibrosis in CD samples. We modelled a CD-like fibrosis phenotype by performing repeated colonic biopsies in mice and analysed the model by histology, type I collagen-targeted positron emission tomography (PET) and global gene expression. We generated a gene set list of essential features of mesenteric fibrosis and compared it to mucosal biopsy datasets from inflammatory bowel disease patients to identify a refined gene set that correlated with clinical outcomes.ResultsMesenteric fibrosis in CD was interconnected to areas of fibrosis in all layers of the intestine, defined as penetrating fibrosis. We found a transcriptional signature of differentially expressed genes enriched in areas of the mesenteric fat of CD subjects with high levels of fibrosis. Mice subjected to repeated colonic biopsies showed penetrating fibrosis as shown by histology, PET imaging and transcriptional analysis. Finally, we composed a composite 24-gene set list that was linked to inflammatory fibroblasts and correlated with treatment response.ConclusionWe linked histopathological and molecular features of CD penetrating fibrosis to a mouse model of repeated biopsy injuries. This experimental system provides an innovative approach for functional investigations of underlying profibrotic mechanisms and therapeutic concepts in CD.
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- 2022
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7. HOIL1 regulates group 2 innate lymphoid cell numbers and type 2 inflammation in the small intestine
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Wood, Matthew J., Marshall, Jeffrey N., Hartley, Victoria L., Liu, Ta-Chiang, Iwai, Kazuhiro, Stappenbeck, Thaddeus S., and MacDuff, Donna A.
- Abstract
Patients with mutations in HOIL1experience a complex immune disorder including intestinal inflammation. To investigate the role of HOIL1in regulating intestinal inflammation, we employed a mouse model of partial HOIL1 deficiency. The ileum of HOIL1-deficient mice displayed features of type 2 inflammation including tuft cell and goblet cell hyperplasia, and elevated expression of Il13, Il5and Il25mRNA. Inflammation persisted in the absence of T and B cells, and bone marrow chimeric mice revealed a requirement for HOIL1 expression in radiation-resistant cells to regulate inflammation. Although disruption of IL-4 receptor alpha (IL4Rα) signaling on intestinal epithelial cells ameliorated tuft and goblet cell hyperplasia, expression of Il5and Il13mRNA remained elevated. KLRG1hiCD90logroup 2 innate lymphoid cells were increased independent of IL4Rα signaling, tuft cell hyperplasia and IL-25 induction. Antibiotic treatment dampened intestinal inflammation indicating commensal microbes as a contributing factor. We have identified a key role for HOIL1, a component of the Linear Ubiquitin Chain Assembly Complex, in regulating type 2 inflammation in the small intestine. Understanding the mechanism by which HOIL1 regulates type 2 inflammation will advance our understanding of intestinal homeostasis and inflammatory disorders and may lead to the identification of new targets for treatment.
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- 2022
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8. Western diet reduces small intestinal intraepithelial lymphocytes via FXR-Interferon pathway
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Hung, Chen-Ting, Ma, Changqing, Panda, Santosh K., Trsan, Tihana, Hodel, Miki, Frein, Jennifer, Foster, Amanda, Sun, Shengxiang, Wu, Hung-Ting, Kern, Justin, Mishra, Richa, Jain, Umang, Ho, Ya-Chi, Colonna, Marco, Stappenbeck, Thaddeus S., and Liu, Ta-Chiang
- Abstract
The prevalence of obesity in the United States has continued to increase over the past several decades. Understanding how diet-induced obesity modulates mucosal immunity is of clinical relevance. We previously showed that consumption of a high fat, high sugar “Western” diet (WD) reduces the density and function of small intestinal Paneth cells, a small intestinal epithelial cell type with innate immune function. We hypothesized that obesity could also result in repressed gut adaptive immunity. Using small intestinal intraepithelial lymphocytes (IEL) as a readout, we found that in non-inflammatory bowel disease (IBD) subjects, high body mass index correlated with reduced IEL density. We recapitulated this in wild type (WT) mice fed with WD. A 4-week WD consumption was able to reduce IEL but not splenic, blood, or bone marrow lymphocytes, and the effect was reversible after another 2 weeks of standard diet (SD) washout. Importantly, WD-associated IEL reduction was not dependent on the presence of gut microbiota, as WD-fed germ-free mice also showed IEL reduction. We further found that WD-mediated Farnesoid X Receptor (FXR) activation in the gut triggered IEL reduction, and this was partially mediated by intestinal phagocytes. Activated FXR signaling stimulated phagocytes to secrete type I IFN, and inhibition of either FXR or type I IFN signaling within the phagocytes prevented WD-mediated IEL loss. Therefore, WD consumption represses both innate and adaptive immunity in the gut. These findings have significant clinical implications in the understanding of how diet modulates mucosal immunity.
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- 2024
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9. Paying attention to minutiae: Strain level differences drive disease etiology
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Newhall, Kevin P. and Stappenbeck, Thaddeus S.
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There is increasing interest in and understanding of the role of fungi in the pathogenesis of complex diseases such as inflammatory bowel disease (IBD). Li et al.1have shown that specific strains of Candida albicanspreferentially produce pro-inflammatory phenotypes that could be related to IBD.
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- 2022
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10. Select symbionts drive high IgA levels in the mouse intestine.
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Zhang, Shanshan, Han, Yi, Schofield, Whitman, Nicosia, Michael, Karell, Paul E., Newhall, Kevin P., Zhou, Julie Y., Musich, Ryan J., Pan, Siyi, Valujskikh, Anna, Sangwan, Naseer, Dwidar, Mohammed, Lu, Qiuhe, and Stappenbeck, Thaddeus S.
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Immunoglobulin A (IgA) is an important factor in maintaining homeostasis at mucosal surfaces, yet luminal IgA levels vary widely. Total IgA levels are thought to be driven by individual immune responses to specific microbes. Here, we found that the prebiotic, pectin oligosaccharide (pec-oligo), induced high IgA levels in the small intestine in a T cell-dependent manner. Surprisingly, this IgA-high phenotype was retained after cessation of pec-oligo treatment, and microbiome transmission either horizontally or vertically was sufficient to retain high IgA levels in the absence of pec-oligo. Interestingly, the bacterial taxa enriched in the overall pec-oligo bacterial community differed from IgA-coated microbes in this same community. Rather, a group of ethanol-resistant microbes, highly enriched for Lachnospiraceae bacterium A2, drove the IgA-high phenotype. These findings support a model of intestinal adaptive immunity in which a limited number of microbes can promote durable changes in IgA directed to many symbionts. [Display omitted] • A pectin-derived prebiotic creates an IgA-high phenotype in mice • The pec-oligo-reshaped microbial community dominantly transmits the IgA-high phenotype • The IgA-high phenotype is driven primarily by CD4
+ T cells in the small intestine • Lachnospiraceae bacterium A2 is enriched in IgA-high mice and likely drives high IgA Zhang et al. demonstrate that long-term dietary intervention with pectin-derived oligosaccharides promotes the ability of specific bacteria, such as Lachnospiraceae bacterium A2, to elevate intestinal IgA that is directed against an array of symbionts. This activity could fit the definition of a keystone species that determines intestinal IgA levels. [ABSTRACT FROM AUTHOR]- Published
- 2023
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11. Atg14protects the intestinal epithelium from TNF-triggered villus atrophy
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Jung, Haerin, Leal-Ekman, J. Steven, Lu, Qiuhe, and Stappenbeck, Thaddeus S.
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ABSTRACTRegulation of intestinal epithelial turnover is a key component of villus maintenance in the intestine. The balance of cell turnover can be perturbed by various extrinsic factors including the cytokine TNF, a cell signaling protein that mediates both proliferative and cytotoxic outcomes. Under conditions of infection and damage, defects in autophagy are associated with TNF-mediated cell death and tissue damage in the intestinal epithelium. However, a direct role of autophagy within the context of enterocyte cell death during homeostasis is lacking. Here, we generated mice lacking ATG14 (autophagy related 14) within the intestinal epithelium [Atg14f/fVil1-Cre (VC)+]. These mice developed spontaneous villus loss and intestinal epithelial cell death within the small intestine. Based on marker studies, the increased cell death in these mice was due to apoptosis. Atg14f/fVC+intestinal epithelial cells demonstrated sensitivity to TNF-triggered apoptosis. Correspondingly, both TNF blocking antibody and genetic deletion of Tnfrsf1a/Tnfr1rescued villus loss and cell death phenotype in Atg14f/fVC+mice. Lastly, we identified a similar pattern of spontaneous villus atrophy and cell death when Rb1cc1/Fip200was conditionally deleted from the intestinal epithelium (Rb1cc1f/fVC+). Overall, these findings are consistent with the hypothesis that factors that control entry into the autophagy pathway are also required during homeostasis to prevent TNF triggered death in the intestine.Abbreviations:ANOVA: analysis of variance; Atg14: autophagy related 14; Atg16l1: autophagy related 16-like 1 (S. cerevisiae); Atg5: autophagy related 5; cCASP3: cleaved CASP3/caspase-3; cCASP8: cleaved CASP8/caspase-8; CHX: cycloheximide; EdU: 5-ethynyl-2´-deoxyuridine thymidine; f/f: flox/flox; H&E: hematoxylin and eosin; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; Nec-1: necrostatin-1; Rb1cc1/Fip200: RB1-inducible coiled-coil 1; Ripk1: receptor (TNFRSF)-interacting serine-threonine kinase 1; Ripk3: receptor (TNFRSF)-interacting serine-threonine kinase 3; Tnfrsf1a/Tnfr1: tumor necrosis factor receptor superfamily, member 1a; Tnf/ Tnfsf1a: tumor necrosis factor; VC: Vil1/villin 1-Cre
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- 2019
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12. Ileal Gene Expression Data from Crohn's Disease Small Bowel Resections Indicate Distinct Clinical Subgroups.
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Potdar, Alka A, Li, Dalin, Haritunians, Talin, VanDussen, Kelli L, Fiorino, Marie F, Liu, Ta-Chiang, Stappenbeck, Thaddeus S, Fleshner, Phillip, Targan, Stephan R, McGovern, Dermot P B, and Bilsborough, Janine
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Background and Aims Heterogeneity in Crohn's disease [CD] provides a challenge for the development of effective therapies. Our goal was to define a unique molecular signature for severe, refractory CD to enable precision therapy approaches to disease treatment and to facilitate earlier intervention in complicated disease. Methods We analysed clinical metadata, genetics, and transcriptomics from uninvolved ileal tissue from CD patients who underwent a single small bowel resection. We determined transcriptional risk scores, cellular signatures, and mechanistic pathways that define patient subsets in refractory CD. Results Within refractory CD, we found three CD patient subgroups [CD1, CD2, and CD3]. Compared with CD1, CD3 was enriched for subjects with increased disease recurrence after first surgery [OR = 6.78, p = 0.04], enhanced occurrence of second surgery [OR = 5.07, p = 0.016], and presence of perianal CD [OR = 3.61, p = 0.036]. The proportion of patients with recurrence-free survival was smaller in CD3 than in CD1 (p = 0.02, median survival time [months] in CD1 = 10 and CD3 = 6). Overlaying differential gene expression between CD1 and CD3 on CD subgroup-associated genetic polymorphisms identified 174 genes representing both genetic and biological differences between the CD subgroups. Pathway analyses using this unique gene signature indicated eukaryotic initiation factor 2 [ eIF2 ] and cyclic adenosine monophosphate [cAMP] signalling to be dominant pathways associated with CD3. Furthermore, the severe, refractory subset, CD3, was associated with a higher transcriptional risk score and enriched with eosinophil and natural killer T [NKT] cell gene signatures. Conclusion We characterized a subset of severe, refractory CD patients who may need more aggressive treatment after first resection and who are likely to benefit from targeted therapy based on their genotype and tissue gene expression signature. [ABSTRACT FROM AUTHOR]
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- 2019
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13. Abnormal Small Intestinal Epithelial Microvilli in Patients With Crohn's Disease.
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VanDussen, Kelli L., Stojmirović, Aleksandar, Li, Katherine, Liu, Ta-Chiang, Kimes, Patrick K., Muegge, Brian D., Simpson, Katherine F., Ciorba, Matthew A., Perrigoue, Jacqueline G., Friedman, Joshua R., Towne, Jennifer E., Head, Richard D., and Stappenbeck, Thaddeus S.
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Background & Aims Crohn disease (CD) presents as chronic and often progressive intestinal inflammation, but the contributing pathogenic mechanisms are unclear. We aimed to identify alterations in intestinal cells that could contribute to the chronic and progressive course of CD. Methods We took an unbiased system-wide approach by performing sequence analysis of RNA extracted from formalin-fixed paraffin-embedded ileal tissue sections from patients with CD (n = 36) and without CD (controls; n = 32). We selected relatively uninflamed samples, based on histology, before gene expression profiling; validation studies were performed using adjacent serial tissue sections. A separate set of samples (3 control and 4 CD samples) was analyzed by transmission electron microscopy. We developed methods to visualize an overlapping modular network of genes dysregulated in the CD samples. We validated our findings using biopsy samples (110 CD samples for gene expression analysis and 54 for histologic analysis) from the UNITI-2 phase 3 trial of ustekinumab for patients with CD and healthy individuals (26 samples used in gene expression analysis). Results We identified gene clusters that were altered in nearly all CD samples. One cluster encoded genes associated with the enterocyte brush border, leading us to investigate microvilli. In ileal tissues from patients with CD, the microvilli were of decreased length and had ultrastructural defects compared with tissues from controls. Microvilli length correlated with expression of genes that regulate microvilli structure and function. Network analysis linked the microvilli cluster to several other down-regulated clusters associated with altered intracellular trafficking and cellular metabolism. Enrichment of a core microvilli gene set also was lower in the UNITI-2 trial CD samples compared with controls; expression of microvilli genes was correlated with microvilli length and endoscopy score and was associated with response to treatment. Conclusions In a transcriptome analysis of formalin-fixed and paraffin-embedded ileal tissues from patients with CD and controls, we associated transcriptional alterations with histologic alterations, such as differences in microvilli length. Decreased microvilli length and decreased expression of the microvilli gene set might contribute to epithelial malfunction and the chronic and progressive disease course in patients with CD. Graphical abstract [ABSTRACT FROM AUTHOR]
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- 2018
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14. Multi-omics of the gut microbial ecosystem in inflammatory bowel diseases
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Lloyd-Price, Jason, Arze, Cesar, Ananthakrishnan, Ashwin N., Schirmer, Melanie, Avila-Pacheco, Julian, Poon, Tiffany W., Andrews, Elizabeth, Ajami, Nadim J., Bonham, Kevin S., Brislawn, Colin J., Casero, David, Courtney, Holly, Gonzalez, Antonio, Graeber, Thomas G., Hall, A. Brantley, Lake, Kathleen, Landers, Carol J., Mallick, Himel, Plichta, Damian R., Prasad, Mahadev, Rahnavard, Gholamali, Sauk, Jenny, Shungin, Dmitry, Vázquez-Baeza, Yoshiki, White, Richard A., Braun, Jonathan, Denson, Lee A., Jansson, Janet K., Knight, Rob, Kugathasan, Subra, McGovern, Dermot P. B., Petrosino, Joseph F., Stappenbeck, Thaddeus S., Winter, Harland S., Clish, Clary B., Franzosa, Eric A., Vlamakis, Hera, Xavier, Ramnik J., and Huttenhower, Curtis
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Inflammatory bowel diseases, which include Crohn’s disease and ulcerative colitis, affect several million individuals worldwide. Crohn’s disease and ulcerative colitis are complex diseases that are heterogeneous at the clinical, immunological, molecular, genetic, and microbial levels. Individual contributing factors have been the focus of extensive research. As part of the Integrative Human Microbiome Project (HMP2 or iHMP), we followed 132 subjects for one year each to generate integrated longitudinal molecular profiles of host and microbial activity during disease (up to 24 time points each; in total 2,965 stool, biopsy, and blood specimens). Here we present the results, which provide a comprehensive view of functional dysbiosis in the gut microbiome during inflammatory bowel disease activity. We demonstrate a characteristic increase in facultative anaerobes at the expense of obligate anaerobes, as well as molecular disruptions in microbial transcription (for example, among clostridia), metabolite pools (acylcarnitines, bile acids, and short-chain fatty acids), and levels of antibodies in host serum. Periods of disease activity were also marked by increases in temporal variability, with characteristic taxonomic, functional, and biochemical shifts. Finally, integrative analysis identified microbial, biochemical, and host factors central to this dysregulation. The study’s infrastructure resources, results, and data, which are available through the Inflammatory Bowel Disease Multi’omics Database (http://ibdmdb.org), provide the most comprehensive description to date of host and microbial activities in inflammatory bowel diseases.
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- 2019
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15. Sa1862 ABNORMAL PANETH CELL PHENOTYPE IN INFLAMMATORY BOWEL DISEASE PATIENTS IS ASSOCIATED WITH DEFECTS IN SECRETORY GRANULE MATURATION SIGNALING.
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Ma, Changqing, Haritunians, Talin, Mujukian, Angela, Hanna, Mary, Li, Dalin, VanDussen, Kelli L., Head, Richard D., Stappenbeck, Thaddeus S., Mcgovern, Dermot P.B., and Liu, Ta-Chiang
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- 2023
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16. Paneth Cell Alterations in the Development and Phenotype of Crohn’s Disease.
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Stappenbeck, Thaddeus S. and McGovern, Dermot P.B.
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Pathogenesis of Crohn’s disease (CD) involves immune and microbial dysregulation, induced by environmental factors in genetically susceptible individuals. There are believed to be multiple subtypes of CD, which contributes to its observed clinical heterogeneity. This concept has been reinforced by recognition of the complexity of the genetic, microbial, immune, and environmental factors that affect risk for CD. Paneth cells mediate immunity and maintain the small intestinal epithelium; defects in activities of these cells have been observed in high proportions of patients with CD, and are associated with a more aggressive CD phenotype. Paneth cells integrate complex genetic, immune, and environmental signals, therefore alterations in their function could lead to different subtypes of CD, as observed in studies in cohorts of primarily European descent. Subtypes of CD associated with Paneth cell function have been observed even among patients from different genetic backgrounds. We discuss genetic susceptibility loci for CD and how these affect Paneth cell activity. [ABSTRACT FROM AUTHOR]
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- 2017
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17. A Common Mechanism Links Activities of Butyrate in the Colon
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Verma, Mohit S., Fink, Michael J., Salmon, Gabriel L., Fornelos, Nadine, Ohara, Takahiro E., Ryu, Stacy H., Vlamakis, Hera, Xavier, Ramnik J., Stappenbeck, Thaddeus S., and Whitesides, George M.
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Two biological activities of butyrate in the colon (suppression of proliferation of colonic epithelial stem cells and inflammation) correlate with inhibition of the activity of histone deacetylases. Cellular and biochemical studies of molecules similar in structure to butyrate, but different in molecular details (functional groups, chain-length, deuteration, oxidation level, fluorination, or degree of unsaturation), demonstrated that these activities were sensitive to molecular structure, and were compatible with the hypothesis that butyrate acts by binding to the Zn2+in the catalytic site of histone deacetylases. Structure–activity relationships drawn from a set of 36 compounds offer a starting point for the design of new compounds targeting the inhibition of histone deacetylases. The observation that butyrate was more potent than other short-chain fatty acids is compatible with the hypothesis that crypts evolved (at least in part), to separate stem cells at the base of crypts from butyrate produced by commensal bacteria.
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- 2018
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18. Microbiota produced indole metabolites disrupt mitochondrial function and inhibit Cryptosporidium parvumgrowth
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Funkhouser-Jones, Lisa J., Xu, Rui, Wilke, Georgia, Fu, Yong, Shriefer, Lawrence A., Makimaa, Heyde, Rodgers, Rachel, Kennedy, Elizabeth A., VanDussen, Kelli L., Stappenbeck, Thaddeus S., Baldridge, Megan T., and Sibley, L. David
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Cryptosporidiosis is a leading cause of life-threatening diarrhea in young children in resource-poor settings. To explore microbial influences on susceptibility, we screened 85 microbiota- associated metabolites for their effects on C. parvumgrowth in vitro. We identify eight inhibitory metabolites in three main classes: secondary bile salts/acids, a vitamin B6precursor, and indoles. Growth restriction of C. parvumby indoles does not depend on the host aryl hydrocarbon receptor (AhR) pathway. Instead, treatment impairs host mitochondrial function and reduces total cellular ATP, as well as directly reducing the membrane potential in the parasite mitosome, a degenerate mitochondria. Oral administration of indoles, or reconstitution of the gut microbiota with indole producing bacteria, delays life cycle progression of the parasite in vitro and reduces severity of C. parvuminfection in mice. Collectively, these findings indicate that microbiota metabolites impair mitochondrial function and contribute to colonization resistance to Cryptosporidiuminfection.
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- 2023
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19. Colitogenic Bacteroides thetaiotaomicron Antigens Access Host Immune Cells in a Sulfatase-Dependent Manner via Outer Membrane Vesicles.
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Hickey, Christina A., Kuhn, Kristine A., Donermeyer, David L., Porter, Nathan T., Jin, Chunsheng, Cameron, Elizabeth A., Jung, Haerin, Kaiko, Gerard E., Wegorzewska, Marta, Malvin, Nicole P., Glowacki, Robert W.P., Hansson, Gunnar C., Allen, Paul M., Martens, Eric C., and Stappenbeck, Thaddeus S.
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Summary Microbes interact with the host immune system via several potential mechanisms. One essential step for each mechanism is the method by which intestinal microbes or their antigens access specific host immune cells. Using genetically susceptible mice ( dnKO ) that develop spontaneous, fulminant colitis, triggered by Bacteroides thetaiotaomicron ( B. theta ), we investigated the mechanism of intestinal microbial access under conditions that stimulate colonic inflammation. B. theta antigens localized to host immune cells through outer membrane vesicles (OMVs) that harbor bacterial sulfatase activity. We deleted the anaerobic sulfatase maturating enzyme (anSME) from B. theta , which is required for post-translational activation of all B. theta sulfatase enzymes. This bacterial mutant strain did not stimulate colitis in dnKO mice. Lastly, access of B. theta OMVs to host immune cells was sulfatase dependent. These data demonstrate that bacterial OMVs and associated enzymes promote inflammatory immune stimulation in genetically susceptible hosts. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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20. Type I Interferons Link Viral Infection to Enhanced Epithelial Turnover and Repair.
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Sun, Lulu, Miyoshi, Hiroyuki, Origanti, Sofia, Nice, Timothy J., Barger, Alexandra C., Manieri, Nicholas A., Fogel, Leslie A., French, Anthony R., Piwnica-Worms, David, Piwnica-Worms, Helen, Virgin, Herbert W., Lenschow, Deborah J., and Stappenbeck, Thaddeus S.
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Summary The host immune system functions constantly to maintain chronic commensal and pathogenic organisms in check. The consequences of these immune responses on host physiology are as yet unexplored, and may have long-term implications in health and disease. We show that chronic viral infection increases epithelial turnover in multiple tissues, and the antiviral cytokines type I interferons (IFNs) mediate this response. Using a murine model with persistently elevated type I IFNs in the absence of exogenous viral infection, the Irgm1 −/− mouse, we demonstrate that type I IFNs act through nonepithelial cells, including macrophages, to promote increased epithelial turnover and wound repair. Downstream of type I IFN signaling, the highly related IFN-stimulated genes Apolipoprotein L9a and b activate epithelial proliferation through ERK activation. Our findings demonstrate that the host immune response to chronic viral infection has systemic effects on epithelial turnover through a myeloid-epithelial circuit. [ABSTRACT FROM AUTHOR]
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- 2015
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21. 963: HIGH FAT DIET-INDUCED MICROVILLUS SHORTENING OF ILEAL ENTEROCYTES IS RESCUED BY ACTIVATION OF PPARα SIGNALING IN MICE.
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Simpson, Katherine, Socha, Jacob J., Muegge, Brian, Kern, Justin, Xiong, Shanshan, Liu, Ta-Chiang, Stappenbeck, Thaddeus S., and VanDussen, Kelli L.
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- 2022
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22. 444: MODULATION OF IGA DEGRADATION AND MUCOSAL ADAPTIVE IMMUNITY BY A NOVEL INTESTINAL SYMBIONT.
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Stappenbeck, Thaddeus S. and Lu, Qiuhe
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- 2022
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23. 20: LRRK2 ACTIVITY MODULATES PANETH CELL FUNCTION IN CROHN'S DISEASE.
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Kern, Justin, Haritunians, Talin, Wang, Xiang, Malique, Atika, Debebe, Anketse, Ballentine, Samuel, Nguyen, Hoang, Vicente, Javier De, Arguello, Annie, Henry, Anastasia, Peter, Inga, Mcgovern, Dermot P.B., Estrada, Anthony, Stappenbeck, Thaddeus S., and Liu, Ta-Chiang
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- 2022
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24. Accounting for reciprocal host–microbiome interactions in experimental science
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Stappenbeck, Thaddeus S. and Virgin, Herbert W.
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Mammals are defined by their metagenome, a combination of host and microbiome genes. This knowledge presents opportunities to further basic biology with translation to human diseases. However, the now-documented influence of the metagenome on experimental results and the reproducibility of in vivomammalian models present new challenges. Here we provide the scientific basis for calling on all investigators, editors and funding agencies to embrace changes that will enhance reproducible and interpretable experiments by accounting for metagenomic effects. Implementation of new reporting and experimental design principles will improve experimental work, speed discovery and translation, and properly use substantial investments in biomedical research.
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- 2016
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25. IL13 activates autophagy to regulate secretion in airway epithelial cells
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Dickinson, John D, Alevy, Yael, Malvin, Nicole P, Patel, Khushbu K, Gunsten, Sean P, Holtzman, Michael J, Stappenbeck, Thaddeus S, and Brody, Steven L
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ABSTRACTCytokine modulation of autophagy is increasingly recognized in disease pathogenesis, and current concepts suggest that type 1 cytokines activate autophagy, whereas type 2 cytokines are inhibitory. However, this paradigm derives primarily from studies of immune cells and is poorly characterized in tissue cells, including sentinel epithelial cells that regulate the immune response. In particular, the type 2 cytokine IL13 (interleukin 13) drives the formation of airway goblet cells that secrete excess mucus as a characteristic feature of airway disease, but whether this process is influenced by autophagy was undefined. Here we use a mouse model of airway disease in which IL33 (interleukin 33) stimulation leads to IL13-dependent formation of airway goblet cells as tracked by levels of mucin MUC5AC (mucin 5AC, oligomeric mucus/gel forming), and we show that these cells manifest a block in mucus secretion in autophagy gene Atg16l1-deficient mice compared to wild-type control mice. Similarly, primary-culture human tracheal epithelial cells treated with IL13 to stimulate mucus formation also exhibit a block in MUC5AC secretion in cells depleted of autophagy gene ATG5(autophagy-related 5) or ATG14(autophagy-related 14) compared to nondepleted control cells. Our findings indicate that autophagy is essential for airway mucus secretion in a type 2, IL13-dependent immune disease process and thereby provide a novel therapeutic strategy for attenuating airway obstruction in hypersecretory inflammatory diseases such as asthma, chronic obstructive pulmonary disease, and cystic fibrosis lung disease. Taken together, these observations suggest that the regulation of autophagy by Th2 cytokines is cell-context dependent.
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- 2016
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26. Unique and redundant functions of NKp46+ ILC3s in models of intestinal inflammation
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Song, Christina, Lee, Jacob S., Gilfillan, Susan, Robinette, Michelle L., Newberry, Rodney D., Stappenbeck, Thaddeus S., Mack, Matthias, Cella, Marina, and Colonna, Marco
- Abstract
Group 3 ILCs (ILC3s) are innate sources of IL-22 and IL-17 and include lymphoid tissue-inducer (LTi)-like and NKp46+ subsets. Both depend on RORγt and aryl hydrocarbon receptor, but NKp46+ILC3s also require Notch and T-bet for their development and are transcriptionally distinct. The extent to which these subsets have unique functions, especially in the context of T cell– and B cell–sufficient mice, remains largely unclear. To investigate the specific function of NKp46+ILC3s among other ILC3 subsets and T cells, we generated mice selectively lacking NKp46+ILC3s or all ILC3s and crossed them to T cell–deficient mice, thus maintaining B cells in all mice. In mice lacking T cells, NKp46+ILC3s were sufficient to promote inflammatory monocyte accumulation in the anti-CD40 innate colitis model through marked production of GM-CSF. In T cell–competent mice, lack of NKp46+ILCs had no impact on control of intestinal C. rodentium infection, whereas lack of all ILC3s partially impaired bacterial control. Thus, NKp46+ILC3s have a unique capacity to promote inflammation through GM-CSF–induced accumulation of inflammatory monocytes, but are superseded by LTi-like ILC3s and T cells in controlling intestinal bacterial infection.
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- 2015
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27. Genetic Variants Synthesize to Produce Paneth Cell Phenotypes That Define Subtypes of Crohn's Disease.
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VanDussen, Kelli L., Liu, Ta-Chiang, Li, Dalin, Towfic, Fadi, Modiano, Nir, Winter, Rachel, Haritunians, Talin, Taylor, Kent D., Dhall, Deepti, Targan, Stephan R., Xavier, Ramnik J., McGovern, Dermot P.B., and Stappenbeck, Thaddeus S.
- Abstract
Background & Aims: Genetic susceptibility loci for Crohn's disease (CD) are numerous, complex, and likely interact with undefined components of the environment. It has been a challenge to link the effects of particular loci to phenotypes of cells associated with pathogenesis of CD, such as Paneth cells. We investigated whether specific phenotypes of Paneth cells associated with particular genetic susceptibility loci can be used to define specific subtypes of CD. Methods: We performed a retrospective analysis of 119 resection specimens collected from patients with CD at 2 separate medical centers. Paneth cell phenotypes were classified as normal or abnormal (with disordered, diminished, diffuse, or excluded granule phenotypes) based on lysozyme-positive secretory granule morphology. To uncover the molecular basis of the Paneth cell phenotypes, we developed methods to determine transcriptional profiles from whole-thickness and laser-capture microdissected, formalin-fixed, paraffin-embedded tissue sections. Results: The proportion of abnormal Paneth cells was associated with the number of CD-associated NOD2 risk alleles. The cumulative number of NOD2 and ATG16L1 risk alleles had an additive effect on the proportion of abnormal Paneth cells. Unsupervised clustering analysis of demographic and Paneth cell data divided patients into 2 principal subgroups, defined by high and low proportions of abnormal Paneth cells. The disordered and diffuse abnormal Paneth cell phenotypes were associated with an altered transcriptional signature of immune system activation. We observed an inverse correlation between abnormal Paneth cells and presence of granuloma. In addition, high proportions of abnormal Paneth cells were associated with shorter time to disease recurrence after surgery. Conclusions: Histologic analysis of Paneth cell phenotypes can be used to divide patients with CD into subgroups with distinct pathognomonic and clinical features. [Copyright &y& Elsevier]
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- 2014
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28. Deletion of p38-alpha mitogen-activated protein kinase within the intestinal epithelium promotes colon tumorigenesis.
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Wakeman, Derek, Schneider, John E., Liu, Jingxia, Wandu, Wambul S., Erwin, Christopher R., Guo, Jun, Stappenbeck, Thaddeus S., and Warner, Brad W.
- Subjects
CARCINOGENESIS ,MITOGEN-activated protein kinase kinase ,INTESTINES ,EPITHELIUM ,COLON cancer treatment ,TUMOR suppressor genes - Abstract
Background: p38-Alpha mitogen-activated protein kinase (p38-MAPK) is a tumor suppressor often mutated in human cancers, but its specific role in colorectal cancer is not completely understood. Previous studies have found that p38-MAPK activity inhibits epithelial proliferation and promotes apoptosis in the intestine. Therefore, we sought to test the hypothesis that intestinal disruption of p38-MAPK would lead to increased tumorigenesis in the colon. Methods: p38-MAPK was deleted in mice within the intestinal epithelium using a tamoxifen-inducible Cre system under control of the villin promoter [villin-Cre ERT2(+), MAPK14(f/f)]. An azoxymethane and dextran sodium sulfate protocol was used to drive intestinal tumor development. Tumor measurements were made using computer software from photographs of excised colon specimens. Results: The number of mice that developed tumors was not statistically different when comparing wild-type mice (7/14) to inducible, intestine epithelial-deleted p38-MAPK (9/11) mice after azoxymethane/dextran sodium sulfate treatment (P = .21). However, the epithelial-deleted p38-MAPK mice developed significantly more tumors (3.7 vs 1.1; P = .008) and nearly 4 times the total tumor burden as wild-type mice (17.4 vs 4.8 mm
2 ; P = .03). Wild-type and epithelial-deleted p38-MAPK groups demonstrated a similar degree of colon inflammation. Conclusion: Deletion of p38-MAPK within the colonic mucosa leads to a hyperplastic state promoting greater tumor development. Because the severity of colitis was not augmented in mice with p38-MAPK deficiency, tumor development is likely mediated by impaired cell cycle regulation within the colonic epithelium. Manipulation of p38-MAPK activity may provide a novel treatment and/or prevention strategy in the management of colorectal cancer, particularly in the setting of inflammatory bowel disease. [ABSTRACT FROM AUTHOR]- Published
- 2012
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29. Commensal Bacteroides Species Induce Colitis in Host-Genotype-Specific Fashion in a Mouse Model of Inflammatory Bowel Disease.
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Bloom, Seth M., Bijanki, Vinieth N., Nava, Gerardo M., Sun, Lulu, Malvin, Nicole P., Donermeyer, David L., Dunne, W. Michael, Allen, Paul M., and Stappenbeck, Thaddeus S.
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BACTEROIDES ,COLITIS ,INFLAMMATORY bowel diseases ,DISEASE susceptibility ,GENETIC mutation ,ENTEROBACTERIACEAE ,ETIOLOGY of diseases ,LABORATORY mice - Abstract
Summary: The intestinal microbiota is important for induction of inflammatory bowel disease (IBD). IBD is associated with complex shifts in microbiota composition, but it is unclear whether specific bacterial subsets induce IBD and, if so, whether their proportions in the microbiota are altered during disease. Here, we fulfilled Koch''s postulates in host-genotype-specific fashion using a mouse model of IBD with human-relevant disease-susceptibility mutations. From screening experiments we isolated common commensal Bacteroides species, introduced them into antibiotic-pretreated mice, and quantitatively reisolated them in culture. The bacteria colonized IBD-susceptible and -nonsusceptible mice equivalently, but induced disease exclusively in susceptible animals. Conversely, commensal Enterobacteriaceae were >100-fold enriched during spontaneous disease, but an Enterobacteriaceae isolate failed to induce disease in antibiotic-pretreated mice despite robust colonization. We thus demonstrate that IBD-associated microbiota alterations do not necessarily reflect underlying disease etiology. These findings establish important experimental criteria and a conceptual framework for understanding microbial contributions to IBD. [Copyright &y& Elsevier]
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- 2011
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30. Paneth Cell Development, Differentiation, and Function: New Molecular Cues.
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Stappenbeck, Thaddeus S.
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- 2009
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31. In vitro expansion and genetic modification of gastrointestinal stem cells in spheroid culture
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Miyoshi, Hiroyuki and Stappenbeck, Thaddeus S
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It is useful to be able to grow enriched populations of stem cells in vitro. Growth of stem cells as tissue spheroids is a key methodology permitting sustainable culture of adult epithelial cells. Gastrointestinal stem cells can be propagated by using conditioned medium from a supportive cell line (L-WRN). This protocol describes how to prepare conditioned medium and how to culture stem cell–enriched epithelial spheroids from the mouse gastrointestine. These spheroids are also amenable to genetic modification with recombinant lentiviruses. This system enables many types of cell biological assays that have been performed with immortalized cell lines to be applied to spheroids. Isolation of epithelial cell units from mice takes up to 2 h, and stem cell–enriched gastrointestinal spheroids are obtained within 3 d. Genetically modified spheroids with lentiviruses can be obtained in 2 weeks.
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- 2013
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32. Western diet induces Paneth cell defects through microbiome alterations and farnesoid X receptor and type I interferon activation.
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Liu, Ta-Chiang, Kern, Justin T., Jain, Umang, Sonnek, Naomi M., Xiong, Shanshan, Simpson, Katherine F., VanDussen, Kelli L., Winkler, Emma S., Haritunians, Talin, Malique, Atika, Lu, Qiuhe, Sasaki, Yo, Storer, Chad, Diamond, Michael S., Head, Richard D., McGovern, Dermot P.B., and Stappenbeck, Thaddeus S.
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Intestinal Paneth cells modulate innate immunity and infection. In Crohn's disease, genetic mutations together with environmental triggers can disable Paneth cell function. Here, we find that a western diet (WD) similarly leads to Paneth cell dysfunction through mechanisms dependent on the microbiome and farnesoid X receptor (FXR) and type I interferon (IFN) signaling. Analysis of multiple human cohorts suggests that obesity is associated with Paneth cell dysfunction. In mouse models, consumption of a WD for as little as 4 weeks led to Paneth cell dysfunction. WD consumption in conjunction with Clostridium spp. increased the secondary bile acid deoxycholic acid levels in the ileum, which in turn inhibited Paneth cell function. The process required excess signaling of both FXR and IFN within intestinal epithelial cells. Our findings provide a mechanistic link between poor diet and inhibition of gut innate immunity and uncover an effect of FXR activation in gut inflammation. [Display omitted] • Diet-induced obesity results in Paneth cell dysfunction in humans and mice • Consumption of western diet leads to Clostridium -mediated deoxycholic acid conversion • Deoxycholic acid activates both FXR and type I IFN pathways • Both FXR and type I IFN pathways are essential in triggering Paneth cell defects Small intestinal Paneth cells are gatekeepers of gut innate immunity. Liu et al. identified a link between consumption of high fat, high sugar diet (western diet; WD) and Paneth cell dysfunction. The mechanisms involve WD-associated microbiota conversion of bile acids, which activate both FXR and type I interferon pathways. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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33. Host genetic susceptibility, dysbiosis, and viral triggers in inflammatory bowel disease
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Sun, Lulu, Nava, Gerardo M, and Stappenbeck, Thaddeus S
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Inflammatory bowel disease (IBD) is thought to occur in genetically susceptible individuals. However, environmental factors, potentially including shifts in commensal microbiota, are also required to trigger disease. This review discusses some of the recent discoveries in host susceptibility and interaction with the microbial environment, and pinpoints key areas for advancement in our understanding of IBD pathogenesis.
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- 2011
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34. Crohn disease: A current perspective on genetics, autophagy and immunity
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Stappenbeck, Thaddeus S., Rioux, John D., Mizoguchi, Atsushi, Saitoh, Tatsuya, Huett, Alan, Darfeuille-Michaud, Arlette, Wileman, Tom, Mizushima, Noboru, Carding, Simon, Parkes, Miles, and Xavier, Ramnik J.
- Abstract
Crohn disease (CD) is a chronic and debilitating inflammatory condition of the gastrointestinal tract.1Prevalence in western populations is 100–150/100,000 and somewhat higher in Ashkenazi Jews. Peak incidence is in early adult life, although any age can be affected and a majority of affected individuals progress to relapsing and chronic disease. Medical treatments rely significantly on empirical corticosteroid therapy and immunosuppression, and intestinal resectional surgery is frequently required. Thus, 80% of patients with CD come to surgery for refractory disease or complications. It is hoped that an improved understanding of pathogenic mechanisms, for example by studying the genetic basis of CD and other forms of inflammatory bowel diseases (IBD), will lead to improved therapies and possibly preventative strategies in individuals identified as being at risk.
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- 2011
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35. Diversity of the autochthonous colonic microbiota
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Nava, Gerardo M. and Stappenbeck, Thaddeus S.
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A longstanding hypothesis in intestinal microbial ecology is that autochthonous microbes (resident) play a role that is distinct from allochthonous microbes (transient microbes in the fecal stream). A challenge has been to identify this pool of microbes. We used laser capture microdissection to collect microbes from the mouse ascending colon. This area contains transverse folds that mimic human intestinal folds and contains a distinct population of intestinal microbes that is associated with the mucosa. Our analysis of bacterial 16S rRNA genes showed that this area was enriched for Lachnospiraceae and Ruminococcaceae. In this addendum, we further compare this community to studies of mucosa-associated microbes in humans. This analysis reveals common phylogenetic groups of bacteria that are present in both mouse and human. However, we found microorganisms at the genus and species levels including Faecalibacterium prausnitzii which appears to be specific for humans. We propose that that examination of the mucosa-associated microbes in wild type and genetically modified mice will be a valuable component to define host microbial interactions that are essential for homeostasis.
- Published
- 2011
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36. Mesenchymal stem cell therapy of intestinal disease are their effects systemic or localized
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Manieri, Nicholas A and Stappenbeck, Thaddeus S
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Stem cell therapy for intestinal diseases is an emerging area in clinical gastroenterology. We will review recent literature regarding mesenchymal stem cells, which have been utilized in preclinical models and are now headed for clinical trials in several gastrointestinal diseases including inflammatory bowel disease.
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- 2011
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37. Deciphering the ‘black box’ of the intestinal stem cell niche taking direction from other systems
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Walker, Monica R and Stappenbeck, Thaddeus S
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Study of developmental signaling pathways suggests that the intestinal stem cell niche regulates the activity of the crypt-based epithelial progenitors during homeostasis and injury states. The cellular origin of these signals, however, remains poorly defined. Here, we examine the current state of knowledge regarding intestinal epithelial progenitor niches and highlight applicable lessons learned from other systems.
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- 2008
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38. Extranuclear sequestration of phospho-Jun N-terminal kinase and distorted villi produced by activated Rac1 in the intestinal epithelium of chimeric mice
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Stappenbeck, Thaddeus S. and Gordon, Jeffrey I.
- Abstract
Previously, we used a genetic mosaic system to conduct an in vivo analysis of the effects of Rac1 activation on the developing intestinal epithelium (Stappenbeck, T. S. and Gordon, J. I. (2000) Development127, 2629-2642). Expression of a constitutively active human Rac1 (Rac1Leu61) in the 129/Sv-derived small intestinal epithelium of C57Bl/6-ROSA26↔129/Sv chimeric mice led to precocious differentiation of some lineages with accompanying alterations in their apical actin. We have now explored the underlying mechanisms. Rac1Leu61 leads to accumulation of the 46 kDa form of phosphorylated Jun N-terminal kinase (p-Jnk) in the apical cytoplasm, but not in the nucleus of E18.5 proliferating and differentiating intestinal epithelial cells. The effect is cell-autonomous, selective for this mitogen-activated protein kinase family member, and accompanied by apical cytoplasmic accumulation of p21-activated kinase. c-Jun, a downstream nuclear target of p-Jnk, does not show evidence of enhanced phosphorylation, providing functional evidence for cytoplasmic sequestration of p-Jnk in Rac1Leu61-expressing epithelium. In adult chimeras, Rac1 activation augments cell proliferation in crypts of Lieberkühn, without a compensatory change in basal apoptosis and produces a dramatic, very unusual widening of villi. These results reveal a novel in vivo paradigm for Rac1 activation involving p-Jnk-mediated signaling at a distinctive extra-nuclear site, with associated alterations in the actin cytoskeleton. They also provide a new perspective about the determinants of small intestinal villus morphogenesis.
- Published
- 2001
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39. Immune System Dysfunction and Autoimmune Disease in Mice Lacking Emk (Par-1) Protein Kinase
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Hurov, Jonathan B., Stappenbeck, Thaddeus S., Zmasek, Christian M., White, Lynn S., Ranganath, Sheila H., Russell, John H., Chan, Andrew C., Murphy, Kenneth M., and Piwnica-Worms, Helen
- Abstract
ABSTRACTEmk is a serine/threonine protein kinase implicated in regulating polarity, cell cycle progression, and microtubule dynamics. To delineate the role of Emk in development and adult tissues, mice lacking Emkwere generated by targeted gene disruption.Emk−/−mice displayed growth retardation and immune cell dysfunction. Although B- and T-cell development were normal, CD4+T cells lacking Emkexhibited a marked upregulation of the memory marker CD44/pgp-1 and produced more gamma interferon and interleukin-4 on stimulation through the T-cell receptor in vitro. In addition, B-cell responses to T-cell-dependent and -independent antigen challenge were altered in vivo. AsEmk−/−animals aged, they developed splenomegaly, lymphadenopathy, membranoproliferative glomerulonephritis, and lymphocytic infiltrates in the lungs, parotid glands and kidneys. Taken together, these results demonstrate that the Emk protein kinase is essential for maintaining immune system homeostasis and that loss of Emk may contribute to autoimmune disease in mammals.
- Published
- 2001
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40. Immune System Dysfunction and Autoimmune Disease in Mice Lacking Emk (Par-1) Protein Kinase
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Hurov, Jonathan B., Stappenbeck, Thaddeus S., Zmasek, Christian M., White, Lynn S., Ranganath, Sheila H., Russell, John H., Chan, Andrew C., Murphy, Kenneth M., and Piwnica-Worms, Helen
- Abstract
Emk is a serine/threonine protein kinase implicated in regulating polarity, cell cycle progression, and microtubule dynamics. To delineate the role of Emk in development and adult tissues, mice lacking Emkwere generated by targeted gene disruption.Emk−/−mice displayed growth retardation and immune cell dysfunction. Although B- and T-cell development were normal, CD4+T cells lacking Emkexhibited a marked upregulation of the memory marker CD44/pgp-1 and produced more gamma interferon and interleukin-4 on stimulation through the T-cell receptor in vitro. In addition, B-cell responses to T-cell-dependent and -independent antigen challenge were altered in vivo. AsEmk−/−animals aged, they developed splenomegaly, lymphadenopathy, membranoproliferative glomerulonephritis, and lymphocytic infiltrates in the lungs, parotid glands and kidneys. Taken together, these results demonstrate that the Emk protein kinase is essential for maintaining immune system homeostasis and that loss of Emk may contribute to autoimmune disease in mammals.
- Published
- 2001
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41. Rac1 mutations produce aberrant epithelial differentiation in the developing and adult mouse small intestine
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Stappenbeck, Thaddeus S. and Gordon, Jeffrey I.
- Abstract
The mouse small intestinal epithelium undergoes continuous renewal throughout life. Previous studies suggest that differentiation of this epithelium is regulated by instructions that are received as cells migrate along crypt-villus units. The nature of the instructions and their intracellular processing remain largely undefined. In this report, we have used genetic mosaic analysis to examine the role of Rac1 GTPase-mediated signaling in controlling differentiation. A constitutively active mutation (Rac1Leu61) or a dominant negative mutation (Rac1Asn17) was expressed in the 129/Sv embryonic stem cell-derived component of the small intestine of C57Bl/6-ROSA26↔129/Sv mice. Rac1Leu61 induces precocious differentiation of members of the Paneth cell and enterocytic lineages in the proliferative compartment of the fetal gut, without suppressing cell division. Forced expression of the dominant negative mutation inhibits epithelial differentiation, without affecting cell division, and slows enterocytic migration along crypt-villus units. The effects produced by Rac1Leu61 or Rac1Asn17 in the 129/Sv epithelium do not spread to adjacent normal C57Bl/6 epithelial cells. These results provide in vivo evidence that Rac1 is involved in the import and intracellular processing of signals that control differentiation of a mammalian epithelium.
- Published
- 2000
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42. Altered Intestinal ACE2 Levels Are Associated With Inflammation, Severe Disease, and Response to Anti-Cytokine Therapy in Inflammatory Bowel Disease.
- Author
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Potdar, Alka A., Dube, Shishir, Naito, Takeo, Li, Katherine, Botwin, Gregory, Haritunians, Talin, Li, Dalin, Casero, David, Yang, Shaohong, Bilsborough, Janine, Perrigoue, Jacqueline G., Denson, Lee A., Daly, Mark, Targan, Stephan R., Fleshner, Phillip, Braun, Jonathan, Kugathasan, Subra, Stappenbeck, Thaddeus S., and McGovern, Dermot P.B.
- Abstract
The host receptor for severe acute respiratory syndrome coronavirus 2, angiotensin-converting enzyme 2 (ACE2), is highly expressed in small bowel (SB). Our aim was to identify factors influencing intestinal ACE2 expression in Crohn's disease (CD), ulcerative colitis (UC), and non–inflammatory bowel disease (IBD) controls. Using bulk RNA sequencing or microarray transcriptomics from tissue samples (4 SB and 2 colonic cohorts; n = 495; n = 387 UC; n = 94 non-IBD), we analyzed the relationship between ACE2 with demographics and disease activity and prognosis. We examined the outcome of anti–tumor necrosis factor and anti–interleukin-12/interleukin-23 treatment on SB and colonic ACE2 expression in 3 clinical trials. Univariate and multivariate regression models were fitted. ACE2 levels were consistently reduced in SB CD and elevated in colonic UC compared with non-IBD controls. Elevated SB ACE2 was also associated with demographic features (age and elevated body mass index) associated with poor coronavirus disease 2019 outcomes. Within CD, SB ACE2 was reduced in patients subsequently developing complicated disease. Within UC, colonic ACE2 was elevated in active disease and in patients subsequently requiring anti–tumor necrosis factor rescue therapy. SB and colonic ACE2 expression in active CD and UC were restored by anti-cytokine therapy, most notably in responders. Reduced SB but elevated colonic ACE2 levels in IBD are associated with inflammation and severe disease, but normalized after anti-cytokine therapy, suggesting compartmentalization of ACE2 -related biology in SB and colonic inflammation. The restoration of ACE2 expression with anti-cytokine therapy might be important in the context of severe acute respiratory syndrome coronavirus 2 infection and potentially explain reports of reduced morbidity from coronavirus disease 2019 in IBD patients treated with anti-cytokines. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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43. Desmoplakin expression and distribution in cultured rat bladder epithelial cells of varying tumorigenic potential
- Author
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Green, Kathleen J., Stappenbeck, Thaddeus S., Noguchi, Sumio, Oyasu, Ryoichi, and Nilles, Laura A.
- Abstract
The expression and distribution of the desmosomal plaque proteins, desmoplakins (DPs) I and II, were studied in nontumorigenic (RBE-8) and a series of tumorigenic (AY34, R-4909, SS-24B, RBTCC-8, and 804G) rat bladder epithelial cell lines. These cell lines ranged from slow-growing papillary transitional cells (AY34) to rapidly metastatic carcinoma cells (RBTCC-8). DPs I and II were shown by immunoblotting and Northern analysis to be present in nontumorigenic RBE-8 cells as well as in all of the tumorigenic cell lines, albeit in differing amounts. Immunofluorescence microscopy revealed striking differences in DP distribution, corresponding in general with increases in tumorigenic potential. Whereas DPs of normal RBE-8 cells and less tumorigenic AY34 cells were localized predominantly at cell interfaces, the more tumorigenic lines exhibited a high proportion of DP in the form of cytoplasmic dots, a distribution reminiscent of that seen in epithelial cells maintained in low levels of extracellular calcium. In 804G cells, which represented the most extreme example of this phenomenon, the majority of DPs were organized as cytoplasmic dots. Electron microscopy revealed intermediate filament (IF)-associated spots in the cytoplasm as well as an elaborate array of IF-associated plaques at the cell-substratum interface. The IF-associated spots in the cytoplasm reacted with anti-DP antibody in immunogold labeling experiments while those at the cell-substratum did not react. In more dense cultures of 804G cells, certain cells stratified and expressed increased amounts of DP followed by the induction of new keratins including those of the skin type. Decreasing extracellular calcium resulted in a rearrangement of DP in each cell line; staining at cell-cell interfaces disappeared and was replaced with a pattern of cytoplasmic dots. These results demonstrate a possible relationship between desmosome assembly and/or maintenance and tumorigenic potential.
- Published
- 1991
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44. Structure of Desmoplakin and Its Association with Intermediate Filaments
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Green, Kathleen J., Stappenbeck, Thaddeus S., Parry, David A.D., and Virata, M. Luisa A.
- Abstract
Desmoplakins (DPs) I and II are two major related proteins located in the desmosomal plaque where they have been proposed to play a role in attaching intermediate filaments (IF) to the inner cell surface. The predicted amino acid sequence of DP was obtained by analysis of overlapping cDNA clones. Computer‐aided analysis suggests that DPI will form a dumbbell‐shaped homodimer, with a central α‐helical coiled coil rod domain of 132 nm and two globular end domains. The DPII molecule is missing 599 residues from the central domain, resulting in a rod about one third the length of DPI. The carboxyl terminus comprises three subdomains each containing almost 5 repeats of a 38 residue repeating motif with a periodicity in acidic and basic residues similar to that found in the rod domain of IF proteins. This suggests a possible mechanism by which these proteins might interact. The amino terminus contains groups of heptad repeats that are predicted to form at least two major α‐helical rich bundles.
- Published
- 1992
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45. Neonatal Mouse Gut Metabolites Influence Cryptosporidium parvumInfection in Intestinal Epithelial Cells
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VanDussen, Kelli L., Funkhouser-Jones, Lisa J., Akey, Marianna E., Schaefer, Deborah A., Ackman, Kevin, Riggs, Michael W., Stappenbeck, Thaddeus S., and Sibley, L. David
- Abstract
Cryptosporidiumsp. occupies a unique intracellular niche that exposes the parasite to both host cell contents and the intestinal lumen, including metabolites from the diet and produced by the microbiota. Both dietary and microbial products change over the course of early development and could contribute to the changes seen in susceptibility to cryptosporidiosis in humans and mice.
- Published
- 2020
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46. A common role for Atg16L1, Atg5, and Atg7 in small intestinal Paneth cells and Crohn disease
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Cadwell, Ken, Patel, Khushbu K., Komatsu, Masaaki, Virgin, Herbert W., and Stappenbeck, Thaddeus S.
- Abstract
Recently identified genetic determinants for enhanced susceptibility to Crohn’s disease (CD) included polymorphisms in the ATG16L1and IRGM1loci suggesting that the autophagy pathway plays a role in the pathogenesis of this disease. We have generated and analyzed three mouse models with diminished expression of autophagy proteins and show how the loss of function of various autophagy components contributes to CD pathogenesis. In the mouse small intestine, the common cellular target of Atg16L1, Atg5, and Atg7 is the Paneth cell, a specialized epithelial cell whose main function is the delivery of antimicrobial factors into the intestinal lumen by production and secretion of its characteristic cytoplasmic granules. Autophagy-deficient Paneth cells exhibited a striking loss of function in this granule exocytosis pathway. Transcriptional analysis revealed a gain of function whereby the gene expression associated with inflammatory responses was increased in autophagy-deficient Paneth cells. Importantly, we validated these findings by analyzing intestinal tissues from CD patients. Similar Paneth cell abnormalities were observed in CD patients homozygous for theATG16L1risk allele. Thus, one role for the autophagy pathway in CD pathogenesis is through selective effects on the biology and specialized properties of Paneth cells.
- Published
- 2009
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47. A Stem-Cell-Derived Platform Enables Complete Cryptosporidium Development In Vitro and Genetic Tractability.
- Author
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Wilke, Georgia, Funkhouser-Jones, Lisa J., Wang, Yi, Ravindran, Soumya, Wang, Qiuling, Beatty, Wandy L., Baldridge, Megan T., VanDussen, Kelli L., Shen, Bang, Kuhlenschmidt, Mark S., Kuhlenschmidt, Theresa B., Witola, William H., Stappenbeck, Thaddeus S., and Sibley, L. David
- Abstract
Despite being a frequent cause of severe diarrheal disease in infants and an opportunistic infection in immunocompromised patients, Cryptosporidium research has lagged due to a lack of facile experimental methods. Here, we describe a platform for complete life cycle development and long-term growth of C. parvum in vitro using "air-liquid interface" (ALI) cultures derived from intestinal epithelial stem cells. Transcriptomic profiling revealed that differentiating epithelial cells grown under ALI conditions undergo profound changes in metabolism and development that enable completion of the parasite life cycle in vitro. ALI cultures support parasite expansion > 100-fold and generate viable oocysts that are transmissible in vitro and to mice, causing infection and animal death. Transgenic parasite lines created using CRISPR/Cas9 were used to complete a genetic cross in vitro , demonstrating Mendelian segregation of chromosomes during meiosis. ALI culture provides an accessible model that will enable innovative studies into Cryptosporidium biology and host interactions. • Air-liquid interface (ALI) cultivation of Cryptosporidium supports robust parasite growth • Both asexual and sexual phases of the parasite complete development in ALI cultures • ALI culture supports the production of de novo oocysts that can trigger an infection in mice • In vitro crossing in ALI cultures opens up forward genetics for Cryptosporidium Wilke et al. describe an air-liquid interface (ALI) cultivation system that permits the efficient growth of C. parvum in vitro. ALI culture supports life cycle completion, thereby generating oocysts that can propagate in vitro , cause infection in mice, and enable in vitro genetic crosses, thus opening this system for future studies. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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48. L-WRN conditioned medium for gastrointestinal epithelial stem cell culture shows replicable batch-to-batch activity levels across multiple research teams.
- Author
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VanDussen, Kelli L., Sonnek, Naomi M., and Stappenbeck, Thaddeus S.
- Abstract
Conditioned medium (CM) derived from engineered cells often facilitates the cost-effective culture of a variety of stem cells. Growing emphasis on the importance of rigor and reproducibility in lab-based science requires development of best practices approaches, including quality control procedures for the assessment of CM batches to ensure reliable interpretation and reproducibility. Here, we tested activity level variations of L-WRN CM, which is produced from an L cell line engineered to secrete W nt3a, R spondin 3, and N oggin into a single CM that is widely used for gastrointestinal stem cell culture. We assessed 14 independent batches of L-WRN CM, produced by 5 laboratories at 3 research institutions, by multiple quantitative assays. We observed highly replicable activity levels among L-WRN CM batches prepared according to a previously published protocol. Quality control assays measuring spheroid growth or mRNA gene marker expression were best able to distinguish the quality L-WRN CM batches, whereas a Wnt reporter assay did not. Thus, we have validated that L-WRN CM activity is highly reproducible over time and between laboratories and have provided guidelines for L-WRN CM quality control testing. These validation procedures and guidelines will benefit experiment replication efforts in stem cell research. Unlabelled Image • Thirteen batches of L-WRN conditioned medium had highly replicable activity levels. • Similar activity levels were observed with medium from 5 laboratory groups. • A Wnt reporter assay did not discriminate quality batches. • Spheroid growth and mRNA marker analysis best discriminated quality batches. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
49. Living and commuting in intestinal crypts
- Author
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Wong, Melissa H., Stappenbeck, Thaddeus S., and Gordon, Jeffrey I.
- Abstract
GASTROENTEROLOGY 1999;116:208-210
- Published
- 1999
- Full Text
- View/download PDF
50. Temporal Regulation of the Bacterial Metabolite Deoxycholate during Colonic Repair Is Critical for Crypt Regeneration.
- Author
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Jain, Umang, Lai, Chin-Wen, Xiong, Shanshan, Goodwin, Victoria M., Lu, Qiuhe, Muegge, Brian D., Christophi, George P., VanDussen, Kelli L., Cummings, Bethany P., Young, Erick, Hambor, John, and Stappenbeck, Thaddeus S.
- Abstract
Summary Colonic wound repair is an orchestrated process, beginning with barrier re-establishment and followed by wound channel formation and crypt regeneration. Elevated levels of prostaglandin E2 (PGE2) promote barrier re-establishment; however, we found that persistently elevated PGE2 hinders subsequent repair phases. The bacterial metabolite deoxycholate (DCA) promotes transition through repair phases via PGE2 regulation. During barrier re-establishment, DCA levels are locally diminished in the wound, allowing enhanced PGE2 production and barrier re-establishment. However, during transition to the wound channel formation phase, DCA levels increase to inhibit PGE2 production and promote crypt regeneration. Altering DCA levels via antibiotic treatment enhances PGE2 levels but impairs wound repair, which is rescued with DCA treatment. DCA acts via its receptor, farnesoid X receptor, to inhibit the enzyme cPLA2 required for PGE2 synthesis. Thus, colonic wound repair requires temporally regulated signals from microbial metabolites to coordinate host-associated signaling cascades. Video Abstract Graphical Abstract Highlights • PGE2 promotes barrier re-establishment but inhibits crypt regeneration in the colon • DCA and PGE2 levels are temporally and reciprocally regulated during colonic repair • DCA acts via its receptor FXR to inhibit PGE2 production and promote crypt regeneration • DCA/FXR inhibits cPLA2 involved in PGE2 synthesis to suppress PGE2 production Jain et al. discover that temporal regulation of a bacterial metabolite, deoxycholate, orchestrates phases of colonic repair. In the first phase, deoxycholate is depleted to allow for barrier re-establishment; it re-emerges in the second phase to promote crypt regeneration. Mechanistically, deoxycholate promotes these transitions by modulating PGE2 levels. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
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