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 Yasui; Mark Williams

doi:10.1136/gutjnl-2012-304067

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 Yasui

Mark Williams

Research output: Contribution to journal › Article › peer-review

87 Citations (Scopus)

37 Downloads (Pure)

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 language	English
Pages (from-to)	610-621
Number of pages	12
Journal	Gut
Volume	63
Issue number	4
Early online date	5 Jul 2013
DOIs	https://doi.org/10.1136/gutjnl-2012-304067
Publication status	Published - 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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10.1136/gutjnl-2012-304067Licence: CC BY

gut_williams
This is an Open Access article distributed in accordance with the terms of the Creative Commons Attribution (CC BY 3.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited. See: http://creativecommons.org/licenses/by/3.0/
Final published version, 4.88 MBLicence: CC BY

Andrea Münsterberg
- A.Munsterberguea.acuk
- School of Biological Sciences - Professor of Developmental Biology
- Cells and Tissues - Member
Person: Research Group Member, Academic, Teaching and Research
Anastasia Sobolewski
- A.Sobolewskiuea.acuk
- School of Chemistry, Pharmacy and Pharmacology - Lecturer in Pharmacology
- Pharmacology and Biological Chemistry - Member
Person: Research Group Member, Academic, Teaching and Research
Mark Williams
- M.R.Williamsuea.acuk
- School of Biological Sciences - Associate Professor
- Norwich Institute for Healthy Aging - Member
Person: Research Centre Member, Academic, Teaching and Research

Epigenetic modulation of Wnt signalling in the ageing human intestinal epithelium: consequences to tissue
Williams, M. (Principal Investigator)
Biotechnology and Biological Sciences Research Council
1/10/08 → 30/09/11
Project: Research
Functional analysis of integrated Wnt signals for self-renewal of the human colonic epithelium
Williams, M. (Principal Investigator) & Münsterberg, A. (Co-Investigator)
Biotechnology and Biological Sciences Research Council
1/12/06 → 31/12/09
Project: Research

Cite this

Reynolds, A., Wharton, N., Parris, A., Mitchell, E., Sobolewski, A., Kam, C., Bigwood, L., El Hadi, A., Münsterberg, A., Lewis, M., Speakman, C., Stebbings, W., Wharton, R., Sargen, K., Tighe, R., Jamieson, C., Hernon, J., Kapur, S., Oue, N., ... Williams, M. (2014). Canonical Wnt signals combined with suppressed TGFβ/BMP pathways promote renewal of the native human colonic epithelium. Gut, 63(4), 610-621. https://doi.org/10.1136/gutjnl-2012-304067

@article{8206ac93838e45398945e233b063b8ea,

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

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. ",

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",

author = "Amy Reynolds and Natalia Wharton and Alyson Parris and Esther Mitchell and Anastasia Sobolewski and Christy Kam and Loren Bigwood and \{El Hadi\}, Ahmed and Andrea M{\"u}nsterberg and Michael Lewis and Christopher Speakman and William Stebbings and Richard Wharton and Kevin Sargen and Richard Tighe and Crawford Jamieson and James Hernon and Sandeep Kapur and Naohide Oue and Wataru Yasui and Mark Williams",

note = "This is an Open Access article distributed in accordance with the terms of the Creative Commons Attribution (CC BY 3.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited. See: http://creativecommons.org/licenses/by/3.0/",

year = "2014",

month = apr,

doi = "10.1136/gutjnl-2012-304067",

language = "English",

volume = "63",

pages = "610--621",

journal = "Gut",

issn = "0017-5749",

publisher = "BMJ Publishing Group",

number = "4",

}

Reynolds, A, Wharton, N, Parris, A, Mitchell, E, Sobolewski, A, Kam, C, Bigwood, L, El Hadi, A, Münsterberg, A, Lewis, M, Speakman, C, Stebbings, W, Wharton, R, Sargen, K, Tighe, R, Jamieson, C, Hernon, J, Kapur, S, Oue, N, Yasui, W & Williams, M 2014, 'Canonical Wnt signals combined with suppressed TGFβ/BMP pathways promote renewal of the native human colonic epithelium', Gut, vol. 63, no. 4, pp. 610-621. https://doi.org/10.1136/gutjnl-2012-304067

TY - JOUR

T1 - Canonical Wnt signals combined with suppressed TGFβ/BMP pathways promote renewal of the native human colonic epithelium

AU - Reynolds, Amy

AU - Wharton, Natalia

AU - Parris, Alyson

AU - Mitchell, Esther

AU - Sobolewski, Anastasia

AU - Kam, Christy

AU - Bigwood, Loren

AU - El Hadi, Ahmed

AU - Münsterberg, Andrea

AU - Lewis, Michael

AU - Speakman, Christopher

AU - Stebbings, William

AU - Wharton, Richard

AU - Sargen, Kevin

AU - Tighe, Richard

AU - Jamieson, Crawford

AU - Hernon, James

AU - Kapur, Sandeep

AU - Oue, Naohide

AU - Yasui, Wataru

AU - Williams, Mark

N1 - This is an Open Access article distributed in accordance with the terms of the Creative Commons Attribution (CC BY 3.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited. See: http://creativecommons.org/licenses/by/3.0/

PY - 2014/4

Y1 - 2014/4

N2 - 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.

AB - 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.

KW - Adult

KW - Aged

KW - Aged, 80 and over

KW - Bone Morphogenetic Proteins

KW - Cell Differentiation

KW - Cell Movement

KW - Cell Proliferation

KW - Colon

KW - Humans

KW - In Situ Hybridization

KW - Intestinal Mucosa

KW - Microscopy, Confocal

KW - Middle Aged

KW - Regeneration

KW - Signal Transduction

KW - Stem Cells

KW - Transforming Growth Factor beta

KW - Wnt Signaling Pathway

U2 - 10.1136/gutjnl-2012-304067

DO - 10.1136/gutjnl-2012-304067

M3 - Article

C2 - 23831735

SN - 0017-5749

VL - 63

SP - 610

EP - 621

JO - Gut

JF - Gut

IS - 4

ER -

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

Abstract

Keywords

Access to Document

Profiles

Andrea Münsterberg

Anastasia Sobolewski

Mark Williams

Projects

Epigenetic modulation of Wnt signalling in the ageing human intestinal epithelium: consequences to tissue

Functional analysis of integrated Wnt signals for self-renewal of the human colonic epithelium

Cite this