Intestinal Enteroids Model Guanylate Cyclase C-Dependent Secretion Induced by Heat-Stable Enterotoxins

Pattison, Amanda M.; Blomain, Erik; Merlino, Dante J.; Wang, Fang; Crissey, Mary Ann S.; Kraft, Crystal; Rappaport, Jeff; Snook, Adam E.; Lynch, John P.

doi:10.1128/iai.00639-16

Cited by 28 publications

(40 citation statements)

References 43 publications

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“…At the molecular level, it is known that the downstream regulators of this pathway, cGMP and PKG, can enhance wild-type and mutant CFTR channel function and trafficking ( Golin-Bisello et al, 2005 ; Leier et al, 2012 ; Poschet et al, 2007 ; Vaandrager et al, 1997 ). Previous studies of mouse derived intestinal organoids showed that agonists of PKG are effective in enhancing CFTR-mediated organoid swelling, in the absence of agonists of the canonical regulation of CFTR, such as PKA ( Pattison et al, 2016 ; Picciotto et al, 1992 ; Seidler et al, 1997 ; Vaandrager et al, 1997 ). Our studies confirm the functional significance of this regulatory pathway in murine colon, and further, our findings suggest that PKG-mediated phosphorylation exerts a ‘priming’ effect on the canonical PKA-mediated activation of CFTR.…”

Section: Discussionmentioning

confidence: 99%

SLC6A14, an amino acid transporter, modifies the primary CF defect in fluid secretion

Ahmadi

Xia

et al. 2018

eLife

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The severity of intestinal disease associated with Cystic Fibrosis (CF) is variable in the patient population and this variability is partially conferred by the influence of modifier genes. Genome-wide association studies have identified SLC6A14, an electrogenic amino acid transporter, as a genetic modifier of CF-associated meconium ileus. The purpose of the current work was to determine the biological role of Slc6a14, by disrupting its expression in CF mice bearing the major mutation, F508del. We found that disruption of Slc6a14 worsened the intestinal fluid secretion defect, characteristic of these mice. In vitro studies of mouse intestinal organoids revealed that exacerbation of the primary defect was associated with reduced arginine uptake across the apical membrane, with aberrant nitric oxide and cyclic GMP-mediated regulation of the major CF-causing mutant protein. Together, these studies highlight the role of this apical transporter in modifying cellular nitric oxide levels, residual function of the major CF mutant and potentially, its promise as a therapeutic target.

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Section: Discussionmentioning

confidence: 99%

SLC6A14, an amino acid transporter, modifies the primary CF defect in fluid secretion

Ahmadi

Xia

et al. 2018

eLife

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“…Should the desired relief still not be achieved then newer prescription agents are available with different mechanisms. These agents include: lubiprostone, a chloride channel type 2 activator and prostaglandin analog 27 ; linaclotide, a GC-C agonist and analog of Escherichia coli heat-stable enterotoxin (STa) 28 , 29 ; and plecanatide, a GC-C agonist and analog of human uroguanylin. 9 Lubiprostone should be administered twice daily with food.…”

Section: Discussionmentioning

confidence: 99%

Randomized clinical trial: efficacy and safety of plecanatide in the treatment of chronic idiopathic constipation

DeMicco

Barrow

Hickey

et al. 2017

Therap Adv Gastroenterol

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Background:Plecanatide, with the exception of a single amino acid replacement, is identical to human uroguanylin and is approved in the United States for adults with chronic idiopathic constipation (CIC). This double-blind, placebo-controlled, phase III study evaluated the efficacy and safety of plecanatide versus placebo in CIC.Methods:Adults meeting modified Rome III CIC criteria were randomized to plecanatide 3 mg (n = 443), 6 mg (n = 449), or placebo (n = 445). Patients recorded bowel movement (BM) characteristics [including spontaneous BMs (SBMs) and complete SBMs (CSBMs)] and rated CIC symptoms in daily electronic diaries. The primary endpoint was the percentage of durable overall CSBM responders (weekly responders for ⩾9 of 12 treatment weeks, including ⩾3 of the last 4 weeks). Weekly responders had ⩾3 CSBMs/week and an increase of ⩾1 CSBM from baseline for the same week.Results:A significantly greater percentage of durable overall CSBM responders resulted with each plecanatide dose compared with placebo (3 mg = 20.1%; 6 mg = 20.0%; placebo = 12.8%; p = 0.004 each dose). Over the 12 weeks, plecanatide significantly improved stool consistency and stool frequency. Significant increases in mean weekly SBMs and CSBMs began in week 1 and were maintained through week 12 in plecanatide-treated patients. Adverse events were mostly mild/moderate, with diarrhea being the most common (3 mg = 3.2%; 6 mg = 4.5%; placebo = 1.3%).Conclusions:Plecanatide resulted in a significantly greater percentage of durable overall CSBM responders and improved stool frequency and secondary endpoints. Plecanatide was well tolerated; the most common AE, diarrhea, occurred in a small number of patients.[ClinicalTrials.gov identifier: NCT02122471]

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“…Intestinal organoid culture Small intestinal organoids derived from GC-C knockout mice were a generous gift from the laboratory of Scott Waldman and were isolated as described previously 21 . Organoids were maintained in 50 µl Matrigel droplets (Corning) in wells of a 24-well plate containing 650 µl IntestiCult mouse organoid growth medium (StemCell Technologies cat# 06005) at 37°C/5% CO2.…”

Section: Cellular Cgmp Assaymentioning

confidence: 99%

Conflicts with diarrheal pathogens trigger rapid evolution of an intestinal signaling axis

Carey

Apple

Hilbert

et al. 2020

Preprint

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The pathogenesis of infectious diarrheal diseases is largely attributed to enterotoxin proteins that disrupt intestinal water absorption, causing severe dehydration. Despite profound health consequences, the impacts of diarrhea-causing microbes on the evolutionary history of host species are largely unknown. We investigated patterns of genetic variation in mammalian Guanylate Cyclase-C (GC-C), an intestinal receptor frequently targeted by bacterial enterotoxins, to determine how hosts might adapt in response to diarrheal infections. Under normal conditions, GC-C interacts with endogenous guanylin peptides to promote water secretion in the intestine, but signaling can be hijacked by bacterially-encoded heat-stable enterotoxins (STa) during infection, which leads to overstimulation of GC-C and diarrhea. Phylogenetic analysis in mammals revealed evidence of recurrent positive selection in the GC-C ligand-binding domain in primates and bats, consistent with selective pressures to evade interactions with STa. Using in vitro assays and transgenic intestinal organoids to model STa-mediated diarrhea, we show that GC-C diversification in these lineages results in substantial variation in toxin susceptibility. In bats, we observe a unique pattern of compensatory coevolution in the endogenous GC-C ligand uroguanylin, reflecting intense bouts of positive selection at the receptor-ligand interface. These findings demonstrate control of water physiology as a previously unrecognized interface for genetic conflict and reveal diarrheal pathogens as a source of selective pressure among diverse mammals.

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Intestinal Enteroids Model Guanylate Cyclase C-Dependent Secretion Induced by Heat-Stable Enterotoxins

Cited by 28 publications

References 43 publications

SLC6A14, an amino acid transporter, modifies the primary CF defect in fluid secretion

SLC6A14, an amino acid transporter, modifies the primary CF defect in fluid secretion

Randomized clinical trial: efficacy and safety of plecanatide in the treatment of chronic idiopathic constipation

Conflicts with diarrheal pathogens trigger rapid evolution of an intestinal signaling axis

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