31 results on '"McGeough, Matthew D."'
Search Results
2. Author Correction: JT002, a small molecule inhibitor of the NLRP3 inflammasome for the treatment of autoinflammatory disorders
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Ambrus-Aikelin, Geza, Takeda, Katsuyuki, Joetham, Anthony, Lazic, Milos, Povero, Davide, Santini, Angelina M., Pranadinata, Rama, Johnson, Casey D., McGeough, Matthew D., Beasley, Federico C., Stansfield, Ryan, McBride, Christopher, Trzoss, Lynnie, Hoffman, Hal M., Feldstein, Ariel E., Stafford, Jeffrey A., Veal, James M., Bain, Gretchen, and Gelfand, Erwin W.
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- 2023
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3. Hepatocyte pyroptosis and release of inflammasome particles induce stellate cell activation and liver fibrosis
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Gaul, Susanne, Leszczynska, Aleksandra, Alegre, Fernando, Kaufmann, Benedikt, Johnson, Casey D, Adams, Leon A, Wree, Alexander, Damm, Georg, Seehofer, Daniel, Calvente, Carolina J, Povero, Davide, Kisseleva, Tatiana, Eguchi, Akiko, McGeough, Matthew D, Hoffman, Hal M, Pelegrin, Pablo, Laufs, Ulrich, and Feldstein, Ariel E
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Biomedical and Clinical Sciences ,Clinical Sciences ,Digestive Diseases ,Liver Disease ,Chronic Liver Disease and Cirrhosis ,2.1 Biological and endogenous factors ,Aetiology ,Inflammatory and immune system ,Oral and gastrointestinal ,Good Health and Well Being ,Animals ,Caspase 1 ,Hepatic Stellate Cells ,Hepatocytes ,Humans ,Inflammasomes ,Interleukin-1beta ,Liver Cirrhosis ,Mice ,Mice ,Inbred NOD ,NLR Family ,Pyrin Domain-Containing 3 Protein ,Non-alcoholic Fatty Liver Disease ,Protein Translocation Systems ,Pyroptosis ,Reactive Oxygen Species ,NLRP3 ,Inflammasome ,Specks ,ASC ,Fibrosis ,NASH ,Liver ,Public Health and Health Services ,Gastroenterology & Hepatology ,Clinical sciences - Abstract
Background & aimsIncreased hepatocyte death contributes to the pathology of acute and chronic liver diseases. However, the role of hepatocyte pyroptosis and extracellular inflammasome release in liver disease is unknown.MethodsWe used primary mouse and human hepatocytes, hepatocyte-specific leucine 351 to proline Nlrp3KICreA mice, and GsdmdKO mice to investigate pyroptotic cell death in hepatocytes and its impact on liver inflammation and damage. Extracellular NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) inflammasomes were isolated from mutant NLRP3-YFP HEK cells and internalisation was studied in LX2 and primary human hepatic stellate cells. We also examined a cohort of 154 adult patients with biopsy-proven non-alcoholic fatty liver disease (Sir Charles Gairdner Hospital, Nedlands, Western Australia).ResultsWe demonstrated that primary mouse and human hepatocytes can undergo pyroptosis upon NLRP3 inflammasome activation with subsequent release of NLRP3 inflammasome proteins that amplify and perpetuate inflammasome-driven fibrogenesis. Pyroptosis was inhibited by blocking caspase-1 and gasdermin D activation. The activated form of caspase-1 was detected in the livers and in serum from patients with non-alcoholic steatohepatitis and correlated with disease severity. Nlrp3KICreA mice showed spontaneous liver fibrosis under normal chow diet, and increased sensitivity to liver damage and inflammation after treatment with low dose lipopolysaccharide. Mechanistically, hepatic stellate cells engulfed extracellular NLRP3 inflammasome particles leading to increased IL-1β secretion and α-smooth muscle actin expression. This effect was abrogated when cells were pre-treated with the endocytosis inhibitor cytochalasin B.ConclusionsThese results identify hepatocyte pyroptosis and release of inflammasome components as a novel mechanism to propagate liver injury and liver fibrosis development.Lay summaryOur findings identify a novel mechanism of inflammation in the liver. Experiments in cell cultures, mice, and human samples show that a specific form of cell death, called pyroptosis, leads to the release of complex inflammatory particles, the NLRP3 inflammasome, from inside hepatocytes into the extracellular space. From there they are taken up by other cells and thereby mediate inflammatory and pro-fibrogenic stress signals. The discovery of this mechanism may lead to novel treatments for chronic liver diseases in the future.
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- 2021
4. Mutations in topoisomerase IIβ result in a B cell immunodeficiency.
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Broderick, Lori, Yost, Shawn, Li, Dong, McGeough, Matthew D, Booshehri, Laela M, Guaderrama, Marisela, Brydges, Susannah D, Kucharova, Karolina, Patel, Niraj C, Harr, Margaret, Hakonarson, Hakon, Zackai, Elaine, Cowell, Ian G, Austin, Caroline A, Hügle, Boris, Gebauer, Corinna, Zhang, Jianguo, Xu, Xun, Wang, Jian, Croker, Ben A, Frazer, Kelly A, Putnam, Christopher D, and Hoffman, Hal M
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B-Lymphocytes ,Animals ,Mice ,Knockout ,Humans ,Mice ,Saccharomyces cerevisiae ,DNA Topoisomerases ,Type II ,Cell Differentiation ,Mutation ,Female ,Male ,Primary Immunodeficiency Diseases - Abstract
B cell development is a highly regulated process involving multiple differentiation steps, yet many details regarding this pathway remain unknown. Sequencing of patients with B cell-restricted immunodeficiency reveals autosomal dominant mutations in TOP2B. TOP2B encodes a type II topoisomerase, an essential gene required to alleviate topological stress during DNA replication and gene transcription, with no previously known role in B cell development. We use Saccharomyces cerevisiae, and knockin and knockout murine models, to demonstrate that patient mutations in TOP2B have a dominant negative effect on enzyme function, resulting in defective proliferation, survival of B-2 cells, causing a block in B cell development, and impair humoral function in response to immunization.
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- 2019
5. Cutting Edge: Targeting Epithelial ORMDL3 Increases, Rather than Reduces, Airway Responsiveness and Is Associated with Increased Sphingosine-1-Phosphate
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Miller, Marina, Tam, Arvin B, Mueller, James L, Rosenthal, Peter, Beppu, Andrew, Gordillo, Ruth, McGeough, Matthew D, Vuong, Christine, Doherty, Taylor A, Hoffman, Hal M, Niwa, Maho, and Broide, David H
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Asthma ,Lung ,2.1 Biological and endogenous factors ,Aetiology ,Respiratory ,Animals ,Disease Models ,Animal ,Lysophospholipids ,Membrane Proteins ,Mice ,Mice ,Inbred C57BL ,Mice ,Knockout ,Polymerase Chain Reaction ,Respiratory Hypersensitivity ,Sphingosine ,Immunology - Abstract
In this study, we used cre-lox techniques to generate mice selectively deficient in ORMDL3 in airway epithelium (Ormdl3Δ2-3/Δ2-3/CC10) to simulate an inhaled therapy that effectively inhibited ORMDL3 expression in the airway. In contrast to the anticipated reduction in airway hyperresponsiveness (AHR), OVA allergen-challenged Ormdl3Δ2-3/Δ2-3/CC10 mice had a significant increase in AHR compared with wild-type mice. Levels of airway inflammation, mucus, fibrosis, and airway smooth muscle were no different in Ormdl3Δ2-3/Δ2-3/CC10 and wild-type mice. However, levels of sphingosine-1-phosphate (S1P) were significantly increased in Ormdl3Δ2-3/Δ2-3/CC10 mice as well as in airway epithelial cells in which ORMDL3 was inhibited with small interfering RNA. Incubation of S1P with airway smooth muscle cells significantly increased contractility. Overall, Ormdl3Δ2-3/Δ2-3/CC10 mice exhibit increased allergen-induced AHR independent of inflammation and associated with increased S1P generation. These studies raise concerns for inhaled therapies that selectively and effectively inhibit ORMDL3 in airway epithelium in asthma.
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- 2017
6. GSDMB induces an asthma phenotype characterized by increased airway responsiveness and remodeling without lung inflammation
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Das, Sudipta, Miller, Marina, Beppu, Andrew K, Mueller, James, McGeough, Matthew D, Vuong, Christine, Karta, Maya R, Rosenthal, Peter, Chouiali, Fazila, Doherty, Taylor A, Kurten, Richard C, Hamid, Qutayba, Hoffman, Hal M, and Broide, David H
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Genetics ,Lung ,Asthma ,Aetiology ,2.1 Biological and endogenous factors ,Respiratory ,Airway Remodeling ,Animals ,Antigens ,Dermatophagoides ,Arachidonate 5-Lipoxygenase ,Bronchial Hyperreactivity ,Cells ,Cultured ,Collagen ,Cytokines ,Epithelial Cells ,Humans ,Matrix Metalloproteinase 9 ,Mice ,Transgenic ,Neoplasm Proteins ,Phenotype ,RNA ,Messenger ,Respiratory Mucosa ,GSDMB ,asthma ,airway-hyperresponsiveness ,remodeling ,inflammation - Abstract
Gasdermin B (GSDMB) on chromosome 17q21 demonstrates a strong genetic linkage to asthma, but its function in asthma is unknown. Here we identified that GSDMB is highly expressed in lung bronchial epithelium in human asthma. Overexpression of GSDMB in primary human bronchial epithelium increased expression of genes important to both airway remodeling [TGF-β1, 5-lipoxygenase (5-LO)] and airway-hyperresponsiveness (AHR) (5-LO). Interestingly, hGSDMBZp3-Cre mice expressing increased levels of the human GSDMB transgene showed a significant spontaneous increase in AHR and a significant spontaneous increase in airway remodeling, with increased smooth muscle mass and increased fibrosis in the absence of airway inflammation. In addition, hGSDMBZp3-Cre mice showed increases in the same remodeling and AHR mediators (TGF-β1, 5-LO) observed in vitro in GSDMB-overexpressing epithelial cells. GSDMB induces TGF-β1 expression via induction of 5-LO, because knockdown of 5-LO in epithelial cells overexpressing GSDMB inhibited TGF-β1 expression. These studies demonstrate that GSDMB, a gene highly linked to asthma but whose function in asthma is previously unknown, regulates AHR and airway remodeling without airway inflammation through a previously unrecognized pathway in which GSDMB induces 5-LO to induce TGF-β1 in bronchial epithelium.
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- 2016
7. NF-κB Restricts Inflammasome Activation via Elimination of Damaged Mitochondria
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Zhong, Zhenyu, Umemura, Atsushi, Sanchez-Lopez, Elsa, Liang, Shuang, Shalapour, Shabnam, Wong, Jerry, He, Feng, Boassa, Daniela, Perkins, Guy, Ali, Syed Raza, McGeough, Matthew D, Ellisman, Mark H, Seki, Ekihiro, Gustafsson, Asa B, Hoffman, Hal M, Diaz-Meco, Maria T, Moscat, Jorge, and Karin, Michael
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Adaptor Proteins ,Signal Transducing ,Animals ,Heat-Shock Proteins ,Inflammasomes ,Interleukin-1beta ,Lipopolysaccharides ,Macrophages ,Mice ,Mitochondria ,NF-kappa B p50 Subunit ,Reactive Oxygen Species ,Sequestosome-1 Protein ,Ubiquitin-Protein Ligases ,Biological Sciences ,Medical and Health Sciences ,Developmental Biology - Abstract
Nuclear factor κB (NF-κB), a key activator of inflammation, primes the NLRP3-inflammasome for activation by inducing pro-IL-1β and NLRP3 expression. NF-κB, however, also prevents excessive inflammation and restrains NLRP3-inflammasome activation through a poorly defined mechanism. We now show that NF-κB exerts its anti-inflammatory activity by inducing delayed accumulation of the autophagy receptor p62/SQSTM1. External NLRP3-activating stimuli trigger a form of mitochondrial (mt) damage that is caspase-1- and NLRP3-independent and causes release of direct NLRP3-inflammasome activators, including mtDNA and mtROS. Damaged mitochondria undergo Parkin-dependent ubiquitin conjugation and are specifically recognized by p62, which induces their mitophagic clearance. Macrophage-specific p62 ablation causes pronounced accumulation of damaged mitochondria and excessive IL-1β-dependent inflammation, enhancing macrophage death. Therefore, the "NF-κB-p62-mitophagy" pathway is a macrophage-intrinsic regulatory loop through which NF-κB restrains its own inflammation-promoting activity and orchestrates a self-limiting host response that maintains homeostasis and favors tissue repair.
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- 2016
8. Mutation of EpCAM leads to intestinal barrier and ion transport dysfunction
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Kozan, Philip A, McGeough, Matthew D, Peña, Carla A, Mueller, James L, Barrett, Kim E, Marchelletta, Ronald R, and Sivagnanam, Mamata
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Medicinal and Biomolecular Chemistry ,Chemical Sciences ,Digestive Diseases ,Women's Health ,Genetics ,2.1 Biological and endogenous factors ,Animals ,Antigens ,Neoplasm ,Cell Adhesion Molecules ,Cell Line ,Epithelial Cell Adhesion Molecule ,Gene Knockdown Techniques ,Intestinal Mucosa ,Ion Transport ,Mice ,Mice ,Knockout ,Mutation ,Permeability ,RNA Interference ,RNA ,Small Interfering ,Congenital tufting enteropathy ,EpCAM ,Ion transport ,Barrier function ,Immunology ,Medicinal and biomolecular chemistry - Abstract
UnlabelledCongenital tufting enteropathy (CTE) is a devastating diarrheal disease seen in infancy that is typically associated with villous changes and the appearance of epithelial tufts. We previously found mutations in epithelial cell adhesion molecule (EpCAM) to be causative in CTE. We developed a knock-down cell model of CTE through transfection of an EpCAM shRNA construct into T84 colonic epithelial cells to elucidate the in vitro role of EpCAM in barrier function and ion transport. Cells with EpCAM deficiency exhibited decreased electrical resistance, increased permeability, and decreased ion transport. Based on mutations in CTE patients, an in vivo mouse model was developed, with tamoxifen-inducible deletion of exon 4 in Epcam resulting in mutant protein with decreased expression. Tamoxifen treatment of Epcam (Δ4/Δ4) mice resulted in pathological features of villous atrophy and epithelial tufts, similar to those in human CTE patients, within 4 days post induction. Epcam (Δ4/Δ4) mice also showed decreased expression of tight junctional proteins, increased permeability, and decreased ion transport in the intestines. Taken together, these findings reveal mechanisms that may underlie disease in CTE.Key messagesKnock-down EpCAM cell model of congenital tufting enteropathy was developed. In vivo inducible mouse model was developed resulting in mutant EpCAM protein. Cells with EpCAM deficiency demonstrated barrier and ion transport dysfunction. Tamoxifen-treated Epcam (Δ4/Δ4) mice demonstrated pathological features. Epcam (Δ4/Δ4) mice showed improper barrier function and ion transport.
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- 2015
9. Independent roles of the priming and the triggering of the NLRP3 inflammasome in the heart
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Toldo, Stefano, Mezzaroma, Eleonora, McGeough, Matthew D, Peña, Carla A, Marchetti, Carlo, Sonnino, Chiara, Van Tassell, Benjamin W, Salloum, Fadi N, Voelkel, Norbert F, Hoffman, Hal M, and Abbate, Antonio
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Cardiovascular ,Heart Disease ,Aetiology ,2.1 Biological and endogenous factors ,Animals ,Cardiomyopathies ,Carrier Proteins ,Caspase 1 ,Disease Models ,Animal ,Inflammasomes ,Lipopolysaccharides ,Mice ,Transgenic ,NLR Family ,Pyrin Domain-Containing 3 Protein ,Cardiomyopathy ,Caspase-1 ,Cryopyrinopathy ,Inflammasome ,NLRP3 ,Priming - Abstract
AimsThe NLRP3 inflammasome is activated in the ischaemic heart promoting caspase-1 activation, inflammation, and cell death. Ischaemic injury establishes both a priming signal (transcription of inflammasome components) and a trigger (NLRP3 activation). Whether NLRP3 activation, without priming, induces cardiac dysfunction and/or failure is unknown. The aim of this study was to assess the independent and complementary roles of the priming and the triggering signals in the heart, in the absence of ischaemia or myocardial injury.Methods and resultsWe used mice with mutant NLRP3 (constitutively active), NLRP3-A350V, under the control of tamoxifen-driven expression of the Cre recombinase (Nlrp3-A350V/CreT mice). The mice were treated for 10 days with tamoxifen before measuring the activity of caspase-1, the effector enzyme in the inflammasome. Tamoxifen treatment induced the inflammasome in the spleen but not in the heart, despite expression of the mutant NLRP3-A350V. The components of the inflammasome were significantly less expressed in the heart compared with the spleen. Subclinical low-dose lipopolysaccharide (LPS; 2 mg/kg) in Nlrp3-A350V/CreT mice induced the expression of the components of the inflammasome (priming), measured using real-time PCR and western blot, leading to the formation of an active inflammasome (caspase-1 activation) in the heart and LV systolic dysfunction while low-dose LPS was insufficient to induce LV systolic dysfunction in wild-type mice (all P < 0.01 for mutant vs. wild-type mice).ConclusionThe signalling pathway governing the inflammasome formation in the heart requires a priming signal in order for an active NLRP3 to induce caspase-1 activation and LV dysfunction.
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- 2015
10. NLRP3 inflammasome activation is required for fibrosis development in NAFLD
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Wree, Alexander, McGeough, Matthew D, Peña, Carla A, Schlattjan, Martin, Li, Hongying, Inzaugarat, Maria Eugenia, Messer, Karen, Canbay, Ali, Hoffman, Hal M, and Feldstein, Ariel E
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Medicinal and Biomolecular Chemistry ,Chemical Sciences ,Hepatitis ,Liver Disease ,Digestive Diseases ,Chronic Liver Disease and Cirrhosis ,Nutrition ,2.1 Biological and endogenous factors ,Aetiology ,Oral and gastrointestinal ,Inflammatory and immune system ,Animals ,Carrier Proteins ,Collagen Type I ,Collagen Type I ,alpha 1 Chain ,Female ,Fibrosis ,Humans ,Inflammasomes ,Liver ,Male ,Mice ,Transgenic ,NLR Family ,Pyrin Domain-Containing 3 Protein ,Non-alcoholic Fatty Liver Disease ,RNA ,Messenger ,NLRP3 ,Inflammation ,Liver fibrosis ,NASH ,Steatoheptatitis ,Immunology ,Medicinal and biomolecular chemistry - Abstract
NLR inflammasomes, caspase 1 activation platforms critical for processing key pro-inflammatory cytokines, have been implicated in the development of nonalcoholic fatty liver disease (NAFLD). As the direct role of the NLRP3 inflammasome remains unclear, we tested effects of persistent NLRP3 activation as a contributor to NAFLD development and, in particular, as a modulator of progression from benign hepatic steatosis to steatohepatitis during diet-induced NAFLD. Gain of function tamoxifen-inducible Nlrp3 knock-in mice allowing for in vivo temporal control of NLRP3 activation and loss of function Nlrp3 knockout mice were placed on short-term choline-deficient amino acid-defined (CDAA) diet, to induce isolated hepatic steatosis or long-term CDAA exposure, to induce severe steatohepatitis and fibrosis, respectively. Expression of NLRP3 associated proteins was assessed in liver biopsies of a well-characterized group of patients with the full spectrum of NAFLD. Nlrp3(-/-) mice were protected from long-term feeding CDAA-induced hepatomegaly, liver injury, and infiltration of activated macrophages. More importantly, Nlrp3(-/-) mice showed marked protection from CDAA-induced liver fibrosis. After 4 weeks on CDAA diet, wild-type (WT) animals showed isolated hepatic steatosis while Nlrp3 knock-in mice showed severe liver inflammation, with increased infiltration of activated macrophages and early signs of liver fibrosis. In the liver samples of patients with NAFLD, inflammasome components were significantly increased in those patients with nonalcoholic steatohepatitis (NASH) when compared to those with non-NASH NAFLD with mRNA levels of pro-IL1 beta correlated to levels of COL1A1. Our study uncovers a crucial role for the NLRP3 inflammasome in the development of NAFLD. These findings may lead to novel therapeutic strategies aimed at halting the progression of hepatic steatosis to the more severe forms of this disease. Key message: Mice with NLRP3 inflammasome loss of function are protected from diet-induced steatohepatitis. NLRP3 inflammasome gain of function leads to early and severe onset of diet-induced steatohepatitis in mice. Patients with severe NAFLD exhibit increased levels of NLRP3 inflammasome components and levels of pro-IL1β mRNA correlate with the expression of COL1A1.
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- 2014
11. ORMDL3 Transgenic Mice Have Increased Airway Remodeling and Airway Responsiveness Characteristic of Asthma
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Miller, Marina, Rosenthal, Peter, Beppu, Andrew, Mueller, James L, Hoffman, Hal M, Tam, Arvin B, Doherty, Taylor A, McGeough, Matthew D, Pena, Carla A, Suzukawa, Maho, Niwa, Maho, and Broide, David H
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Medical Physiology ,Biomedical and Clinical Sciences ,Genetics ,Lung ,Prevention ,Asthma ,2.1 Biological and endogenous factors ,Aetiology ,Respiratory ,Activating Transcription Factor 6 ,Airway Remodeling ,Allergens ,Animals ,Antibody Specificity ,Bronchial Hyperreactivity ,Chemokines ,CC ,Chemokines ,CXC ,Cytokines ,Disease Models ,Animal ,Eosinophils ,Gene Expression ,Gene Order ,Gene Targeting ,Humans ,Immunoglobulin E ,Inflammation ,Membrane Proteins ,Methacholine Chloride ,Mice ,Mice ,Transgenic ,Ovalbumin ,Th2 Cells ,Transgenes ,Unfolded Protein Response ,eIF-2 Kinase ,Immunology ,Biochemistry and cell biology - Abstract
Orosomucoid-like (ORMDL)3 has been strongly linked with asthma in genetic association studies. Because allergen challenge induces lung ORMDL3 expression in wild-type mice, we have generated human ORMDL3 zona pellucida 3 Cre (hORMDL3(zp3-Cre)) mice that overexpress human ORMDL3 universally to investigate the role of ORMDL3 in regulating airway inflammation and remodeling. These hORMDL3(zp3-Cre) mice have significantly increased levels of airway remodeling, including increased airway smooth muscle, subepithelial fibrosis, and mucus. hORMDL3(zp3-Cre) mice had spontaneously increased airway responsiveness to methacholine compared to wild-type mice. This increased airway remodeling was associated with selective activation of the unfolded protein response pathway transcription factor ATF6 (but not Ire1 or PERK). The ATF6 target gene SERCA2b, implicated in airway remodeling in asthma, was strongly induced in the lungs of hORMDL3(zp3-Cre) mice. Additionally, increased levels of expression of genes associated with airway remodeling (TGF-β1, ADAM8) were detected in airway epithelium of these mice. Increased levels of airway remodeling preceded increased levels of airway inflammation in hORMDL3(zp3-Cre) mice. hORMDL3(zp3-Cre) mice had increased levels of IgE, with no change in levels of IgG, IgM, and IgA. These studies provide evidence that ORMDL3 plays an important role in vivo in airway remodeling potentially through ATF6 target genes such as SERCA2b and/or through ATF6-independent genes (TGF-β1, ADAM8).
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- 2014
12. NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice.
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Wree, Alexander, Eguchi, Akiko, McGeough, Matthew D, Pena, Carla A, Johnson, Casey D, Canbay, Ali, Hoffman, Hal M, and Feldstein, Ariel E
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Neutrophils ,Hepatocytes ,Animals ,Mice ,Inbred C57BL ,Humans ,Mice ,Mice ,Mutant Strains ,Hepatitis ,Liver Cirrhosis ,Disease Models ,Animal ,Growth Disorders ,Carrier Proteins ,Receptors ,Interleukin-1 ,Antirheumatic Agents ,Apoptosis ,Point Mutation ,Interleukin 1 Receptor Antagonist Protein ,Hepatic Stellate Cells ,Inflammasomes ,NLR Family ,Pyrin Domain-Containing 3 Protein ,Liver Disease ,Digestive Diseases ,Genetics ,Biotechnology ,Chronic Liver Disease and Cirrhosis ,2.1 Biological and endogenous factors ,Development of treatments and therapeutic interventions ,5.2 Cellular and gene therapies ,Aetiology ,Oral and gastrointestinal ,Good Health and Well Being ,Medical Biochemistry and Metabolomics ,Clinical Sciences ,Immunology ,Gastroenterology & Hepatology - Abstract
UnlabelledInflammasome activation plays a central role in the development of drug-induced and obesity-associated liver disease. However, the sources and mechanisms of inflammasome-mediated liver damage remain poorly understood. Our aim was to investigate the effect of NLRP3 inflammasome activation on the liver using novel mouse models. We generated global and myeloid cell-specific conditional mutant Nlrp3 knock-in mice expressing the D301N Nlrp3 mutation (ortholog of D303N in human NLRP3), resulting in a hyperactive NLRP3. To study the presence and significance of NLRP3-initiated pyroptotic cell death, we separated hepatocytes from nonparenchymal cells and developed a novel flow-cytometry-based (fluorescence-activated cell sorting; FACS) strategy to detect and quantify pyroptosis in vivo based on detection of active caspase 1 (Casp1)- and propidium iodide (PI)-positive cells. Liver inflammation was quantified histologically by FACS and gene expression analysis. Liver fibrosis was assessed by Sirius Red staining and quantitative polymerase chain reaction for markers of hepatic stellate cell (HSC) activation. NLRP3 activation resulted in shortened survival, poor growth, and severe liver inflammation; characterized by neutrophilic infiltration and HSC activation with collagen deposition in the liver. These changes were partially attenuated by treatment with anakinra, an interleukin-1 receptor antagonist. Notably, hepatocytes from global Nlrp3-mutant mice showed marked hepatocyte pyroptotic cell death, with more than a 5-fold increase in active Casp1/PI double-positive cells. Myeloid cell-restricted mutant NLRP3 activation resulted in a less-severe liver phenotype in the absence of detectable pyroptotic hepatocyte cell death.ConclusionsOur data demonstrate that global and, to a lesser extent, myeloid-specific NLRP3 inflammasome activation results in severe liver inflammation and fibrosis while identifying hepatocyte pyroptotic cell death as a novel mechanism of NLRP3-mediated liver damage.
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- 2014
13. Pharmacology of a Potent and Novel Inhibitor of the NOD-Like Receptor Pyrin Domain-Containing Protein 3 (NLRP3) Inflammasome that Attenuates Development of Nonalcoholic Steatohepatitis and Liver Fibrosis
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Povero, Davide, primary, Lazic, Milos, additional, McBride, Christopher, additional, Ambrus-Aikelin, Geza, additional, Stansfield, Ryan, additional, Johnson, Casey D., additional, Santini, Angelina M., additional, Pranadinata, Rama F., additional, McGeough, Matthew D., additional, Stafford, Jeffrey A., additional, Hoffman, Hal M., additional, Feldstein, Ariel E., additional, Veal, James M., additional, and Bain, Gretchen, additional
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- 2023
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14. TNF regulates transcription of NLRP3 inflammasome components and inflammatory molecules in cryopyrinopathies
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McGeough, Matthew D., Wree, Alexander, Haimovich, Ariela, Inzaugarat, Maria E., Johnson, Casey D., Pena, Carla A., Goldbach-Mansky, Raphaela, Broderick, Lori, Feldstein, Ariel E., and Hoffman, Hal M.
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Tumor necrosis factor -- Comparative analysis -- Research ,Cryopyrin-associated periodic syndromes -- Research ,Transcription (Genetics) -- Comparative analysis -- Research ,Health care industry - Abstract
The NLRP3 inflammasome is a protein complex responsible for caspase-1-dependent maturation of the proinflammatory cytokines IL-1[beta] and IL-18. Gain-of-function missense mutations in NLRP3 cause the disease spectrum known as the cryopyrin-associated periodic syndromes (CAPS). In this study, we generated Nlrp3-knockin mice on various KO backgrounds including Il1b/Il18-, caspase-1-, caspase-11- (Casp1/11-), and 7nf-deficient strains. The [Nlrp3.sup.L351P] [Il1b.sup.-/-] [Il18.sup.-/-] mutant mice survived and grew normally until adulthood and, at 6 months of age, exhibited marked splenomegaly and leukophilia. Injection of these mice with low-dose LPS resulted in elevated serum TNF levels compared with [Nlrp3.sup.L351P] [Casp1/11.sup.-/-] mice and [Il1b.sup.-/-] [Il18.sup.-/-] littermates. Treatment of [Nlrp3.sup.A350V] mice with the TNF inhibitor etanercept resulted in all pups surviving to adulthood, with normal body and spleen/body weight ratios. [Nlrp3.sup.A350V] [Tnf.sup.-/-] mice showed a similar phenotypic rescue, with marked reductions in serum IL-1[beta] and IL-18, reduced myeloid inflammatory infiltrate in the skin and spleen, and substantial decreases in splenic mRNA expression of both inflammasome components (Nlrp3, Pycard, pro-Caspf) and pro-cytokines (Il1b, Il18). Likewise, we observed a reduction in the expression of both pro-Casp1 and pro-Il1b in cultured [Nlrp3.sup.A350V] [Tnf.sup.-/-] BM-derived DCs. Our data show that TNF is an important transcriptional regulator of NLRP3 inflammasome components in murine inflammasomopathies. Moreover, these results may have therapeutic implications for CAPS patients with partial responses to IL-1-targeted therapies., Introduction The inflammasome complex is formed by NLRP3, along with the adaptor protein pycard (ASC) and pro-caspase-1, which, when activated by various endogenous and microbe-derived danger signals, directs the autocleavage [...]
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- 2017
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15. The effect of a fennel seed extract on the STAT signaling and intestinal barrier function
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Das, Barun, primary, Rabalais, John, additional, Kozan, Philip, additional, Lu, Tina, additional, Durali, Nassim, additional, Okamoto, Kevin, additional, McGeough, Matthew D., additional, Lee, Beom Jae, additional, Barrett, Kim E., additional, Marchelletta, Ronald, additional, and Sivagnanam, Mamata, additional
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- 2022
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16. Divergence of IL-1, IL-18, and cell death in NLRP3 inflammasomopathies
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Brydges, Susannah D., Broderick, Lori, McGeough, Matthew D., Pena, Carla A., Mueller, James L., and Hoffman, Hal M.
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- 2013
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17. ASK1 inhibition reduces cell death and hepatic fibrosis in an Nlrp3 mutant liver injury model
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Schuster-Gaul, Susanne, primary, Geisler, Lukas Jonathan, additional, McGeough, Matthew D., additional, Johnson, Casey D., additional, Zagorska, Anna, additional, Li, Li, additional, Wree, Alexander, additional, Barry, Vivian, additional, Mikaelian, Igor, additional, Jih, Lily J., additional, Papouchado, Bettina G., additional, Budas, Grant, additional, Hoffman, Hal M., additional, and Feldstein, Ariel E., additional
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- 2020
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18. Enteroids expressing a disease-associated mutant of EpCAM are a model for congenital tufting enteropathy
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Das, Barun, primary, Okamoto, Kevin, additional, Rabalais, John, additional, Kozan, Philip A., additional, Marchelletta, Ronald R., additional, McGeough, Matthew D., additional, Durali, Nassim, additional, Go, Maria, additional, Barrett, Kim E., additional, Das, Soumita, additional, and Sivagnanam, Mamata, additional
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- 2019
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19. Inflammasome-Mediated Disease Animal Models Reveal Roles for Innate but Not Adaptive Immunity
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Brydges, Susannah D., Mueller, James L., McGeough, Matthew D., Pena, Carla A., Misaghi, Amirhossein, Gandhi, Chhavi, Putnam, Chris D., Boyle, David L., Firestein, Gary S., Horner, Anthony A., Soroosh, Pejman, Watford, Wendy T., OʼShea, John J., Kastner, Daniel L., and Hoffman, Hal M.
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- 2009
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20. NLRP3 inflammasome activation in hepatic stellate cells induces murine liver fibrosis
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Inzaugarat, Maria Eugenia, Johnson, Casey D., Holtmann, Theresa Maria, McGeough, Matthew D., Trautwein, Christian, Papouchado, Bettina G., Schwabe, Robert, Hoffman, Hal M., Wree, Alexander, and Feldstein, Ariel E.
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Lipopolysaccharides ,Liver Cirrhosis ,Male ,integumentary system ,Inflammasomes ,Mice, Transgenic ,Article ,Mice, Inbred C57BL ,Liver ,NLR Family, Pyrin Domain-Containing 3 Protein ,Hepatic Stellate Cells ,Animals ,Female ,Myofibroblasts ,Biomarkers - Abstract
The NLRP3 inflammasome plays an important role in liver fibrosis development. However, the mechanisms involved in NLRP3-induced fibrosis are unclear. Our aim was to test the hypothesis that the NLRP3 inflammasome in hepatic stellate cells (HSC) can directly regulate their activation and contribute to liver fibrosis. Primary HSC isolated from WT, Nlrp3(−/−), or Nlrp3(L351PneoR) knock-in crossed to inducible (estrogen receptor Cre - CreT) mice were incubated with LPS and ATP, or 4OH-tamoxifen, respectively. HSC-specific Nlrp3(L351P)-knock-in mice were generated by crossing transgenic mice expressing lecithin retinol acyltransferase (Lrat)-driven Cre and maintained on standard rodent chow for 6 months. Mice were then sacrificed; liver tissue and serum were harvested. Nlrp3 inflammasome activation along with HSC phenotype and fibrosis were assessed by RT-PCR, Western blot, FACS, ELISA, immunofluorescence and immunohistochemistry. Stimulated WT HSC displayed increased levels of NLRP3 inflammasome-induced ROS production and Cathepsin B activity, accompanied by an upregulation of mRNA and protein levels of fibrotic makers, an effect abrogated in Nlrp3(−/−) HSC. Nlrp3(L351P) CreT HSC also showed elevated mRNA and protein expression of fibrotic markers 24h after inflammasome activation induced with 4OH-tamoxifen. Protein and mRNA expression levels of fibrotic markers were also found to be increased in isolated HSC and whole liver tissue from Nlrp3(L351P) Lrat Cre mice compared to WT. Liver sections from 24 week-old Nlrp(L351P) Lrat Cre mice showed fibrotic changes with increased αSMA and desmin positive cells and collagen deposition, independent of inflammatory infiltrates; these changes were also observed after LPS challenge in 8 week-old Nlrp(L351P) Lrat Cre mice. CONCLUSION: Our results highlight a direct role for the NLRP3 inflammasome in the activation of HSC directly triggering liver fibrosis.
- Published
- 2019
21. Increased Neutrophil Secretion Induced by NLRP3 Mutation Links the Inflammasome to Azurophilic Granule Exocytosis
- Author
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Johnson, Jennifer L., primary, Ramadass, Mahalakshmi, additional, Haimovich, Ariela, additional, McGeough, Matthew D., additional, Zhang, Jinzhong, additional, Hoffman, Hal M., additional, and Catz, Sergio D., additional
- Published
- 2017
- Full Text
- View/download PDF
22. Interleukin-6 is a Marker of Inflammation with No Direct Role in Inflammasome-Mediated Mouse Models1
- Author
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McGeough, Matthew D., Pena, Carla A., Mueller, James L., Pociask, Derek A., Broderick, Lori, Hoffman, Hal M., and Brydges, Susannah D.
- Subjects
Mice, Knockout ,Mice, 129 Strain ,Inflammasomes ,Interleukin-6 ,Interleukin-1beta ,Reproducibility of Results ,Mice, Transgenic ,Article ,Cryopyrin-Associated Periodic Syndromes ,Immunophenotyping ,Mice, Inbred C57BL ,Disease Models, Animal ,Mice ,Gene Targeting ,NLR Family, Pyrin Domain-Containing 3 Protein ,Animals ,Gene Knock-In Techniques ,Inflammation Mediators ,Carrier Proteins ,Biomarkers - Abstract
IL-6 is a known downstream target of IL-1β and is consistently increased in serum from patients with NLRP3 inflammasome-mediated conditions. Therefore, IL-6 could be a therapeutic target in the treatment of IL-1β-provoked inflammation. IL-6 was increased in serum with accompanying neutrophilia in tissues of an inducible mouse model of Muckle-Wells syndrome. However, an IL-6-null background failed to provide phenotypic rescue and did not significantly impact inflammatory cytokine levels. In a second model of IL-1β-driven inflammation, NLRP3 activation by monosodium urate crystals similarly increased IL-6. Consistent with our Muckle-Wells syndrome model, ablation of IL-6 did not impact an acute neutrophilic response in this in vivo evaluation of gouty arthritis. Taken together, our results indicate that IL-6 is a reliable marker of inflammation, with no direct role in inflammasome-mediated disease.
- Published
- 2012
23. Independent roles of the priming and the triggering of the NLRP3 inflammasome in the heart
- Author
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Toldo, Stefano, primary, Mezzaroma, Eleonora, additional, McGeough, Matthew D., additional, Peña, Carla A., additional, Marchetti, Carlo, additional, Sonnino, Chiara, additional, Van Tassell, Benjamin W., additional, Salloum, Fadi N., additional, Voelkel, Norbert F., additional, Hoffman, Hal M., additional, and Abbate, Antonio, additional
- Published
- 2014
- Full Text
- View/download PDF
24. Mutation of EpCAM leads to intestinal barrier and ion transport dysfunction
- Author
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Kozan, Philip A., primary, McGeough, Matthew D., additional, Peña, Carla A., additional, Mueller, James L., additional, Barrett, Kim E., additional, Marchelletta, Ronald R., additional, and Sivagnanam, Mamata, additional
- Published
- 2014
- Full Text
- View/download PDF
25. Abstract 14850: Independent Role of the Priming and Triggering of the NLRP3 Inflammasome in the Heart
- Author
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Toldo, Stefano, primary, Mezzaroma, Eleonora, additional, McGeough, Matthew D, additional, Pena, Carla A, additional, Marchetti, Carlo, additional, Sonnino, Chiara, additional, Van Tassell, Benjamin W, additional, Mauro, Adolfo G, additional, Salloum, Fadi N, additional, Hoffman, Hal M, additional, and Abbate, Antonio, additional
- Published
- 2014
- Full Text
- View/download PDF
26. Functional consequences of EpCam mutation in mice and men
- Author
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Mueller, James L., primary, McGeough, Matthew D., additional, Peña, Carla A., additional, and Sivagnanam, Mamata, additional
- Published
- 2014
- Full Text
- View/download PDF
27. Mo1982 Epcam Deficient Mice Demonstrate Intestinal Permeability Defects
- Author
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Sivagnanam, Mamata, primary, Mueller, James L., additional, McGeough, Matthew D., additional, Pena, Carla, additional, and Hoffman, Hal M., additional
- Published
- 2012
- Full Text
- View/download PDF
28. Constitutively Activated NLRP3 Inflammasome Causes Inflammation and Abnormal Skeletal Development in Mice
- Author
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Bonar, Sheri L., primary, Brydges, Susannah D., additional, Mueller, James L., additional, McGeough, Matthew D., additional, Pena, Carla, additional, Chen, Debbie, additional, Grimston, Susan K., additional, Hickman-Brecks, Cynthia L., additional, Ravindran, Soumya, additional, McAlinden, Audrey, additional, Novack, Deborah V., additional, Kastner, Daniel L., additional, Civitelli, Roberto, additional, Hoffman, Hal M., additional, and Mbalaviele, Gabriel, additional
- Published
- 2012
- Full Text
- View/download PDF
29. Epcam Deficient Mice Show Significant Morbidity and Pathology Resembling Congenital Tufting Enteropathy
- Author
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Sivagnanam, Mamata, primary, Mueller, James L., additional, Mcgeough, Matthew D., additional, Pena, Carla, additional, and Hoffman, Hal M., additional
- Published
- 2011
- Full Text
- View/download PDF
30. Enteroids expressing a disease-associated mutant of EpCAM are a model for congenital tufting enteropathy.
- Author
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Das B, Okamoto K, Rabalais J, Kozan PA, Marchelletta RR, McGeough MD, Durali N, Go M, Barrett KE, Das S, and Sivagnanam M
- Subjects
- Animals, Cell Differentiation, Cell Proliferation, Cells, Cultured, Diarrhea, Infantile pathology, Epithelial Cell Adhesion Molecule metabolism, Female, Goblet Cells cytology, Goblet Cells metabolism, Goblet Cells physiology, Intestinal Mucosa metabolism, Intestinal Mucosa pathology, Malabsorption Syndromes pathology, Male, Mice, Mice, Inbred C57BL, Paneth Cells cytology, Paneth Cells metabolism, Paneth Cells physiology, Diarrhea, Infantile genetics, Epithelial Cell Adhesion Molecule genetics, Intestinal Mucosa cytology, Malabsorption Syndromes genetics, Tissue Culture Techniques methods
- Abstract
Congenital tufting enteropathy (CTE) is an autosomal recessive disease characterized by severe intestinal failure in infancy and mutations in the epithelial cell adhesion molecule ( EPCAM ) gene. Previous studies of CTE in mice expressing mutant EpCAM show neonatal lethality. Hence, to study the cellular, molecular, and physiological alterations that result from EpCAM mutation, a tamoxifen-inducible mutant EpCAM enteroid model has been generated. The presence of mutant EpCAM in the model was confirmed at both mRNA and protein levels. Immunofluorescence microscopy demonstrated the reduced expression of mutant EpCAM. Mutant enteroids had reduced budding potential as well as significantly decreased mRNA expression for epithelial lineage markers ( Mucin 2, lysozyme, sucrase-isomaltase ), proliferation marker Ki67, and secretory pathway transcription factors ( Atoh1 , Hnf1b) . Significantly decreased numbers of Paneth and goblet cells were confirmed by staining. These findings were correlated with intestinal tissue from CTE patients and the mutant mice model that had significantly fewer Paneth and goblet cells than in healthy counterparts. FITC-dextran studies demonstrated significantly impaired barrier function in monolayers derived from mutant enteroids compared with control monolayers. In conclusion, we have established an ex vivo CTE model. The role of EpCAM in the budding potential, differentiation, and barrier function of enteroids is noted. Our study establishes new facets of EpCAM biology that will aid in understanding the pathophysiology of CTE and role of EpCAM in health and disease. NEW & NOTEWORTHY Here, we develop a novel ex vivo enteroid model for congenital tufting enteropathy (CTE) based on epithelial cell adhesion molecule ( EPCAM) gene mutations found in patients. With this model we demonstrate the role of EpCAM in maintaining the functional homeostasis of the intestinal epithelium, including differentiation, proliferation, and barrier integrity. This study further establishes a new direction in EpCAM biology that will help in understanding the detailed pathophysiology of CTE and role of EpCAM.
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- 2019
- Full Text
- View/download PDF
31. Functional consequences of EpCam mutation in mice and men.
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Mueller JL, McGeough MD, Peña CA, and Sivagnanam M
- Subjects
- Animals, Antigens, Neoplasm metabolism, Case-Control Studies, Cell Adhesion Molecules metabolism, Cell Movement, Cell Proliferation, Claudins metabolism, Diarrhea, Infantile metabolism, Diarrhea, Infantile pathology, Disease Models, Animal, Epithelial Cell Adhesion Molecule, Exons, Genetic Predisposition to Disease, HEK293 Cells, Humans, Intestinal Absorption, Intestinal Mucosa pathology, Intestines pathology, Malabsorption Syndromes metabolism, Malabsorption Syndromes pathology, Mice, Mice, Knockout, Permeability, Phenotype, Transfection, Antigens, Neoplasm genetics, Cell Adhesion Molecules genetics, Diarrhea, Infantile genetics, Intestinal Mucosa metabolism, Malabsorption Syndromes genetics, Mutation
- Abstract
Congenital tufting enteropathy (CTE) is a severe diarrheal disease of infancy characterized by villous changes and epithelial tufts. We previously identified mutations in epithelial cell adhesion molecule (EpCAM) as the cause of CTE. We developed an in vivo mouse model of CTE based on EpCAM mutations found in patients with the aim to further elucidate the in vivo role of EpCAM and allow for a direct comparison to human CTE. Using Cre-LoxP recombination technology, we generated a construct lacking exon 4 in Epcam. Epcam(Δ4/Δ4) mice and CTE patient intestinal tissue integrity was analyzed by histology using both light immunohistochemistry and electron microscopy. Epcam(Δ4/Δ4) mice demonstrate neonatal lethality and growth retardation with pathological features, including epithelial tufts, enterocyte crowding, altered desmosomes, and intercellular gaps, similar to human CTE patients. Mutant EpCAM protein is present at low levels and is mislocalized in the intestine of Epcam(Δ4/Δ4) mice and CTE patients. Deletion of exon 4 was found to decrease expression of both EpCAM and claudin-7 causing a loss of colocalization, functionally disrupting the EpCAM/claudin-7 complex, a finding for the first time confirmed in CTE patients. Furthermore, compared with unaffected mice, mutation of Epcam leads to enhanced permeability and intestinal cell migration, uncovering underlying disease mechanisms.
- Published
- 2014
- Full Text
- View/download PDF
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