Human Enteroids as a Model of Upper Small Intestinal Ion Transport Physiology and Pathophysiology.

Publisher: W.B. Saunders Country of Publication: United States NLM ID: 0374630 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1528-0012 (Electronic) Linking ISSN: 00165085 NLM ISO Abbreviation: Gastroenterology Subsets: MEDLINE

Publication: Philadelphia, PA : W.B. Saunders
Original Publication: Baltimore.

Cell Differentiation*; Intestine, Small/*metabolism ; Membrane Transport Proteins/*metabolism ; Sodium/*metabolism; Gene Expression Regulation ; Humans ; Hydrogen-Ion Concentration ; Intestinal Secretions/metabolism ; Intestine, Small/pathology ; Intestine, Small/physiopathology ; Ion Transport ; Kinetics ; Membrane Transport Proteins/genetics ; Microscopy, Confocal ; Microscopy, Fluorescence, Multiphoton ; Microscopy, Video ; Time-Lapse Imaging ; Tissue Culture Techniques ; Transduction, Genetic ; Transfection

Background & Aims: Human intestinal crypt-derived enteroids are a model of intestinal ion transport that require validation by comparison with cell culture and animal models. We used human small intestinal enteroids to study neutral Na(+) absorption and stimulated fluid and anion secretion under basal and regulated conditions in undifferentiated and differentiated cultures to show their functional relevance to ion transport physiology and pathophysiology.
Methods: Human intestinal tissue specimens were obtained from an endoscopic biopsy or surgical resections performed at Johns Hopkins Hospital. Crypts were isolated, enteroids were propagated in culture, induced to undergo differentiation, and transduced with lentiviral vectors. Crypt markers, surface cell enzymes, and membrane ion transporters were characterized using quantitative reverse-transcription polymerase chain reaction, immunoblot, or immunofluorescence analyses. We used multiphoton and time-lapse confocal microscopy to monitor intracellular pH and luminal dilatation in enteroids under basal and regulated conditions.
Results: Enteroids differentiated upon withdrawal of WNT3A, yielding decreased crypt markers and increased villus-like characteristics. Na(+)/H(+) exchanger 3 activity was similar in undifferentiated and differentiated enteroids, and was affected by known inhibitors, second messengers, and bacterial enterotoxins. Forskolin-induced swelling was completely dependent on cystic fibrosis transmembrane conductance regulator and partially dependent on Na(+)/H(+) exchanger 3 and Na(+)/K(+)/2Cl(-) cotransporter 1 inhibition in undifferentiated and differentiated enteroids. Increases in cyclic adenosine monophosphate with forskolin caused enteroid intracellular acidification in HCO3(-)-free buffer. Cyclic adenosine monophosphate-induced enteroid intracellular pH acidification as part of duodenal HCO3(-) secretion appears to require cystic fibrosis transmembrane conductance regulator and electrogenic Na(+)/HCO3(-) cotransporter 1.
Conclusions: Undifferentiated or crypt-like, and differentiated or villus-like, human enteroids represent distinct points along the crypt-villus axis; they can be used to characterize electrolyte transport processes along the vertical axis of the small intestine. The duodenal enteroid model showed that electrogenic Na(+)/HCO3(-) cotransporter 1 might be a target in the intestinal mucosa for treatment of secretory diarrheas.
(Copyright © 2016 AGA Institute. Published by Elsevier Inc. All rights reserved.)

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Keywords: Intestinal Organoids; NHE3 Activity; Na(+) Absorption; Stimulated Secretion

0 (Membrane Transport Proteins)
9NEZ333N27 (Sodium)

Date Created: 20151219 Date Completed: 20160706 Latest Revision: 20240610

20240610

PMC4766025

10.1053/j.gastro.2015.11.047

26677983