Canonical Wnt signals combined with suppressed TGFβ/BMP pathways promote renewal of the native human colonic epithelium

Amy Reynolds, Natalia Wharton, Alyson Parris, Esther Mitchell, Anastasia Sobolewski, Christy Kam, Loren Bigwood, Ahmed El Hadi, Andrea Münsterberg, Michael Lewis, Christopher Speakman, William Stebbings, Richard Wharton, Kevin Sargen, Richard Tighe, Crawford Jamieson, James Hernon, Sandeep Kapur, Naohide Oue, Wataru YasuiMark Williams

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Abstract

Background: A defining characteristic of the human intestinal epithelium is that it is the most rapidly renewing tissue in the body. However, the processes underlying tissue renewal and the mechanisms that govern their coordination have proved difficult to study in the human gut.
Objective: To investigate the regulation of stem cell-driven tissue renewal by canonical Wnt and TGFβ/bone morphogenetic protein (BMP) pathways in the native human colonic epithelium.
Design: Intact human colonic crypts were isolated from mucosal tissue samples and placed into 3D culture conditions optimised for steady-state tissue renewal. High affinity mRNA in situ hybridisation and immunohistochemistry were complemented by functional genomic and bioimaging techniques. The effects of signalling pathway modulators on the status of intestinal stem cell biology, crypt cell proliferation, migration, differentiation and shedding were determined.
Results: Native human colonic crypts exhibited distinct activation profiles for canonical Wnt, TGFβ and BMP pathways. A population of intestinal LGR5/OLFM4-positive stem/progenitor cells were interspersed between goblet-like cells within the crypt-base. Exogenous and crypt cell-autonomous canonical Wnt signals supported homeostatic intestinal stem/progenitor cell proliferation and were antagonised by TGFβ or BMP pathway activation. Reduced Wnt stimulation impeded crypt cell proliferation, but crypt cell migration and shedding from the crypt surface were unaffected and resulted in diminished crypts.
Conclusions: Steady-state tissue renewal in the native human colonic epithelium is dependent on canonical Wnt signals combined with suppressed TGFβ/BMP pathways. Stem/progenitor cell proliferation is uncoupled from crypt cell migration and shedding, and is required to constantly replenish the crypt cell population.
Original languageEnglish
Pages (from-to)610-621
Number of pages12
JournalGut
Volume63
Issue number4
Early online date5 Jul 2013
DOIs
Publication statusPublished - Apr 2014

Keywords

  • Adult
  • Aged
  • Aged, 80 and over
  • Bone Morphogenetic Proteins
  • Cell Differentiation
  • Cell Movement
  • Cell Proliferation
  • Colon
  • Humans
  • In Situ Hybridization
  • Intestinal Mucosa
  • Microscopy, Confocal
  • Middle Aged
  • Regeneration
  • Signal Transduction
  • Stem Cells
  • Transforming Growth Factor beta
  • Wnt Signaling Pathway

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