A Novel Human Epithelial Enteroid Model of Necrotizing Enterocolitis

Ares, Guillermo; Buonpane, Christie; Yuan, Carrie; Wood, Douglas R.; Hunter, Catherine J.

doi:10.3791/59194

Cited by 20 publications

(11 citation statements)

References 17 publications

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“…Necrotizing enterocolitis (NEC) is a devastating intestinal disease of premature infants (4,5,21). Despite decades of research, the complex pathophysiology is incompletely understood, and despite advances in neonatal care, the mortality remains high (21).…”

Section: Introductionmentioning

confidence: 99%

ROCK1 inhibitor stabilizes E-cadherin and improves barrier function in experimental necrotizing enterocolitis

Buonpane

Yuan

Wood

et al. 2020

American Journal of Physiology-Gastrointestinal and Liver Physi

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Necrotizing enterocolitis (NEC) is a devastating gastrointestinal disease of newborns. Although incompletely understood, NEC is associated with intestinal barrier dysfunction. E-cadherin, an adherens junction, is a protein complex integral in maintaining normal barrier homeostasis. Rho-associated protein kinase-1 (ROCK1) is a kinase that regulates the E-cadherin complex, and p120-catenin is a subunit of the E-cadherin complex that has been implicated in stabilizing the cadherin complex at the plasma membrane. We hypothesized that E-cadherin is decreased in NEC and that inhibition of ROCK1 would protect against adherens junction disruption. To investigate this, a multimodal approach was used: In vitro Caco-2 model of NEC (LPS/TNFα), rap pup model (hypoxia + bacteria-containing formula), and human intestinal samples. E-cadherin was decreased in NEC compared with controls, with relocalization from the cell border to an intracellular location. ROCK1 exhibited a time-dependent response to disease, with increased early expression in NEC and decreased expression at later time points and disease severity. Administration of ROCK1 inhibitor (RI) resulted in preservation of E-cadherin expression at the cell border, preservation of intestinal villi on histological examination, and decreased apoptosis. ROCK1 upregulation in NEC led to decreased association of E-cadherin to p120 and increased intestinal permeability. RI helped maintain the stability of the E-cadherin-p120 complex, leading to improved barrier integrity and protection from experimental NEC. NEW & NOTEWORTHY This paper is the first to describe the effect of ROCK1 on E-cadherin expression in the intestinal epithelium and the protective effects of ROCK inhibitor on E-cadherin stability in necrotizing enterocolitis.

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Section: Introductionmentioning

confidence: 99%

ROCK1 inhibitor stabilizes E-cadherin and improves barrier function in experimental necrotizing enterocolitis

Buonpane

Yuan

Wood

et al. 2020

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…23 Ares et al have also reported a human epithelial model of NEC that involves culturing human organoids after isolation of intestinal stem cells from patients undergoing bowel resection. 37 The authors demonstrate that administration of LPS in the growth media of organoids leads to changes in histology, genetic and protein expression, as well as an inflammatory response resembling human NEC including increased mRNA and protein expression of TLR4. 37 Development of NEC is preceded by changes in the intestinal microbial population including reduced diversity and colonization by specific species.…”

Section: Neonatal Intestinal Organoids As Ex Vivo Models Of Necmentioning

confidence: 99%

“…37 The authors demonstrate that administration of LPS in the growth media of organoids leads to changes in histology, genetic and protein expression, as well as an inflammatory response resembling human NEC including increased mRNA and protein expression of TLR4. 37 Development of NEC is preceded by changes in the intestinal microbial population including reduced diversity and colonization by specific species. 38 Hence, it is essential to have a clear understanding of the role of microbes in the development of the neonatal intestine, as well as in pathogenesis of NEC.…”

Section: Neonatal Intestinal Organoids As Ex Vivo Models Of Necmentioning

confidence: 99%

Necrotizing Enterocolitis: State of the Art in Translating Experimental Research to the Bedside

Ganji

Lee

et al. 2019

Eur J Pediatr Surg

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Necrotizing enterocolitis (NEC) is a devastating intestinal disease that continues to have high morbidity and mortality among preterm neonates, despite medical advancements in neonatology and neonatal care. To investigate the pathogenesis of the disease and explore novel form of treatment, a variety of experimental models of NEC have been developed and used by various investigators. These experimental models range from in vitro evaluation of intestinal epithelial cells and intestinal organoids to in vivo models of the disease in neonatal mice, rats, and piglets. Most recently, human-derived intestinal organoids have also been developed and investigated. In this review, we will briefly discuss these experimental models and the contributions that they have made to our understanding of NEC. We will also point to the ischemia/reperfusion (I/R) model of intestinal injury which has been used as an indirect model of NEC by some investigators. Advancements in laboratory research into this devastating disease have continued to expand our knowledge on the pathogenesis and prevention of NEC as well as the effectiveness of therapeutic options for management of this severe disease.

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“…The journey from the crypt to detachment from the tip takes approximately 4 to 5 days (Vachon et al, 2000; van der Flier and Clevers, 2009). Crypt cells from the human small intestine have been successfully cultured as primary monolayer cells (Browning and Trier, 1969;Panja, 2000;Benoit et al, 2010;Beaulieu and Ménard, 2012) as well as three-dimensional organoids (enteroids) as an in vitro model for the evaluation of intestinal physiology and diseases including the evaluation of major histocompatibility complex class II regulation (Wang et al, 2018;Wosen et al, 2019), interaction of the enterocytes with pathogenic microbiota (In et al, 2019;Stewart et al, 2020), modeling infectious diarrheal diseases (Kovbasnjuk et al, 2013;Foulke-Abel et al, 2014, elucidation of pathogenesis of intestinal diseases such as inflammatory bowel disease (Rees et al, 2019) and necrotizing enterocolitis (Senger et al, 2018;Ares et al, 2019), and the elucidation of the cell and molecular pathways controlling stem cell maturation into enterocytes (Das et al, 2015;Mahe et al, 2015;Schilderink et al, 2016). As of this writing, there are no reports defining the expression and activity of drug metabolizing enzymes in human enteroids.…”

Section: Stem Cell-derived Modelsmentioning

confidence: 99%

In Vitro Human Cell–Based Experimental Models for the Evaluation of Enteric Metabolism and Drug Interaction Potential of Drugs and Natural Products

2020

Drug Metab Dispos

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Elements of key enteric drug metabolism and disposition pathways are reviewed to aid the assessment of the applicability of current cell-based enteric experimental systems for the evaluation of enteric metabolism and drug interaction potential. Enteric nuclear receptors include vitamin D receptor, constitutive androstane receptor, pregnane X receptor, farnesoid X receptor, liver X receptor, aryl hydrocarbon receptor, and peroxisome proliferator-activated receptor. Enteric drug metabolizing enzyme pathways include both cytochrome P450 (P450) and non-P450 drug metabolizing enzymes based on gene expression, proteomics, and activity. Both uptake and efflux transporters are present in the small intestine, with P-glycoprotein found to be responsible for most drug-drug and food-drug interactions. The cell-based in vitro enteric systems reviewed are 1) immortalized cell line model: the human colon adenocarcinoma (Caco-2) cells; 2) human stem cell-derived enterocyte models: stem cell enteric systems, either from intestinal crypt cells or induced pluripotent stem cells; and 3) primary cell models: human intestinal slices, cryopreserved human enterocytes, permeabilized cofactor-supplemented (MetMax) cryopreserved human enterocytes, and cryopreserved human intestinal mucosa. The major deficiency with both immortalized cell lines and stem cell-derived enterocytes is that drug metabolizing enzyme activities, although they are detectable, are substantially lower than those for the intestinal mucosa in vivo. Human intestine slices, cryopreserved human enterocytes, MetMax cryopreserved human enterocytes, and cryopreserved human intestinal mucosa retain robust enteric drug metabolizing enzyme activity and represent appropriate models for the evaluation of metabolism and metabolism-dependent drug interaction potential of orally administered xenobiotics including drugs, botanical products, and dietary supplements. SIGNIFICANCE STATEMENT Enteric drug metabolism plays an important role in the bioavailability and metabolic fate of orally administered drugs as well as in enteric drugdrug and food-drug interactions. The current status of key enteric drug metabolism and disposition pathways and in vitro human cell-based enteric experimental systems for the evaluation of the metabolism and drug interaction potential of orally administered substances is reviewed.

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A Novel Human Epithelial Enteroid Model of Necrotizing Enterocolitis

Cited by 20 publications

References 17 publications

ROCK1 inhibitor stabilizes E-cadherin and improves barrier function in experimental necrotizing enterocolitis

ROCK1 inhibitor stabilizes E-cadherin and improves barrier function in experimental necrotizing enterocolitis

Necrotizing Enterocolitis: State of the Art in Translating Experimental Research to the Bedside

In Vitro Human Cell–Based Experimental Models for the Evaluation of Enteric Metabolism and Drug Interaction Potential of Drugs and Natural Products

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