Reg4(+) deep crypt secretory cells function as epithelial niche for Lgr5(+) stem cells in colon

Nobuo Sasaki; Norman Sachs; Kay Wiebrands; Saskia I. J. Ellenbroek; Arianna Fumagalli; Anna Lyubimova; Harry Begthel; Maaike van den Born; Johan H. van Es; Wouter R. Karthaus; Vivian S. W. Li; Carmen López Iglesias; Peter Peters; Jacco van Rheenen; Alexander van Oudenaarden; Hans Clevers

doi:10.1073/pnas.1607327113

Reg4(+) deep crypt secretory cells function as epithelial niche for Lgr5(+) stem cells in colon

Nobuo Sasaki, Norman Sachs, Kay Wiebrands, Saskia I. J. Ellenbroek, Arianna Fumagalli, Anna Lyubimova, Harry Begthel, Maaike van den Born, Johan H. van Es, Wouter R. Karthaus, Vivian S. W. Li, Carmen López Iglesias, Peter Peters, Jacco van Rheenen, Alexander van Oudenaarden, Hans Clevers^*

^*Corresponding author for this work

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

Abstract

Leucine-rich repeat-containing G-protein coupled receptor 5-positive (Lgr5(+)) stem cells reside at crypt bottoms of the small and large intestine. Small intestinal Paneth cells supply Wnt3, EGF, and Notch signals to neighboring Lgr5(+) stem cells. Whereas the colon lacks Paneth cells, deep crypt secretory (DCS) cells are intermingled with Lgr5(+) stem cells at crypt bottoms. Here, we report regenerating isletderived family member 4 (Reg4) as a marker of DCS cells. To investigate a niche function, we eliminatedDCS cells by using the diphtheriatoxin receptor gene knocked into themurine Reg4 locus. Ablation of DCS cells results in loss of stem cells from colonic crypts and disrupts gut homeostasis and colon organoid growth. In agreement, sorted Reg4(+) DCS cells promote organoid formation of single Lgr5(+) colon stem cells. DCS cells can be massively produced from Lgr5(+) colon stem cells in vitro by combined Notch inhibition and Wnt activation. We conclude that Reg4(+) DCS cells serve as Paneth cell equivalents in the colon crypt niche.

Original language	English
Pages (from-to)	E5399-E5407
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	113
Issue number	37
DOIs	https://doi.org/10.1073/pnas.1607327113
Publication status	Published - 13 Sept 2016

Keywords

intestinal stem cell
niche
Lgr5
Reg4
deep crypt secretory cells

Access to Document

10.1073/pnas.1607327113

Cite this

Sasaki, N., Sachs, N., Wiebrands, K., Ellenbroek, S. I. J., Fumagalli, A., Lyubimova, A., Begthel, H., van den Born, M., van Es, J. H., Karthaus, W. R., Li, V. S. W., López Iglesias, C., Peters, P., van Rheenen, J., van Oudenaarden, A., & Clevers, H. (2016). Reg4(+) deep crypt secretory cells function as epithelial niche for Lgr5(+) stem cells in colon. Proceedings of the National Academy of Sciences of the United States of America, 113(37), E5399-E5407. https://doi.org/10.1073/pnas.1607327113

@article{ff0ac64ddcb9449aadbd000c2e392fa4,

title = "Reg4(+) deep crypt secretory cells function as epithelial niche for Lgr5(+) stem cells in colon",

abstract = "Leucine-rich repeat-containing G-protein coupled receptor 5-positive (Lgr5(+)) stem cells reside at crypt bottoms of the small and large intestine. Small intestinal Paneth cells supply Wnt3, EGF, and Notch signals to neighboring Lgr5(+) stem cells. Whereas the colon lacks Paneth cells, deep crypt secretory (DCS) cells are intermingled with Lgr5(+) stem cells at crypt bottoms. Here, we report regenerating isletderived family member 4 (Reg4) as a marker of DCS cells. To investigate a niche function, we eliminatedDCS cells by using the diphtheriatoxin receptor gene knocked into themurine Reg4 locus. Ablation of DCS cells results in loss of stem cells from colonic crypts and disrupts gut homeostasis and colon organoid growth. In agreement, sorted Reg4(+) DCS cells promote organoid formation of single Lgr5(+) colon stem cells. DCS cells can be massively produced from Lgr5(+) colon stem cells in vitro by combined Notch inhibition and Wnt activation. We conclude that Reg4(+) DCS cells serve as Paneth cell equivalents in the colon crypt niche.",

keywords = "intestinal stem cell, niche, Lgr5, Reg4, deep crypt secretory cells",

author = "Nobuo Sasaki and Norman Sachs and Kay Wiebrands and Ellenbroek, {Saskia I. J.} and Arianna Fumagalli and Anna Lyubimova and Harry Begthel and {van den Born}, Maaike and {van Es}, {Johan H.} and Karthaus, {Wouter R.} and Li, {Vivian S. W.} and {L{\'o}pez Iglesias}, Carmen and Peter Peters and {van Rheenen}, Jacco and {van Oudenaarden}, Alexander and Hans Clevers",

year = "2016",

month = sep,

day = "13",

doi = "10.1073/pnas.1607327113",

language = "English",

volume = "113",

pages = "E5399--E5407",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

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Sasaki, N, Sachs, N, Wiebrands, K, Ellenbroek, SIJ, Fumagalli, A, Lyubimova, A, Begthel, H, van den Born, M, van Es, JH, Karthaus, WR, Li, VSW, López Iglesias, C , Peters, P, van Rheenen, J, van Oudenaarden, A & Clevers, H 2016, 'Reg4(+) deep crypt secretory cells function as epithelial niche for Lgr5(+) stem cells in colon', Proceedings of the National Academy of Sciences of the United States of America, vol. 113, no. 37, pp. E5399-E5407. https://doi.org/10.1073/pnas.1607327113

TY - JOUR

T1 - Reg4(+) deep crypt secretory cells function as epithelial niche for Lgr5(+) stem cells in colon

AU - Sasaki, Nobuo

AU - Sachs, Norman

AU - Wiebrands, Kay

AU - Ellenbroek, Saskia I. J.

AU - Fumagalli, Arianna

AU - Lyubimova, Anna

AU - Begthel, Harry

AU - van den Born, Maaike

AU - van Es, Johan H.

AU - Karthaus, Wouter R.

AU - Li, Vivian S. W.

AU - López Iglesias, Carmen

AU - Peters, Peter

AU - van Rheenen, Jacco

AU - van Oudenaarden, Alexander

AU - Clevers, Hans

PY - 2016/9/13

Y1 - 2016/9/13

N2 - Leucine-rich repeat-containing G-protein coupled receptor 5-positive (Lgr5(+)) stem cells reside at crypt bottoms of the small and large intestine. Small intestinal Paneth cells supply Wnt3, EGF, and Notch signals to neighboring Lgr5(+) stem cells. Whereas the colon lacks Paneth cells, deep crypt secretory (DCS) cells are intermingled with Lgr5(+) stem cells at crypt bottoms. Here, we report regenerating isletderived family member 4 (Reg4) as a marker of DCS cells. To investigate a niche function, we eliminatedDCS cells by using the diphtheriatoxin receptor gene knocked into themurine Reg4 locus. Ablation of DCS cells results in loss of stem cells from colonic crypts and disrupts gut homeostasis and colon organoid growth. In agreement, sorted Reg4(+) DCS cells promote organoid formation of single Lgr5(+) colon stem cells. DCS cells can be massively produced from Lgr5(+) colon stem cells in vitro by combined Notch inhibition and Wnt activation. We conclude that Reg4(+) DCS cells serve as Paneth cell equivalents in the colon crypt niche.

AB - Leucine-rich repeat-containing G-protein coupled receptor 5-positive (Lgr5(+)) stem cells reside at crypt bottoms of the small and large intestine. Small intestinal Paneth cells supply Wnt3, EGF, and Notch signals to neighboring Lgr5(+) stem cells. Whereas the colon lacks Paneth cells, deep crypt secretory (DCS) cells are intermingled with Lgr5(+) stem cells at crypt bottoms. Here, we report regenerating isletderived family member 4 (Reg4) as a marker of DCS cells. To investigate a niche function, we eliminatedDCS cells by using the diphtheriatoxin receptor gene knocked into themurine Reg4 locus. Ablation of DCS cells results in loss of stem cells from colonic crypts and disrupts gut homeostasis and colon organoid growth. In agreement, sorted Reg4(+) DCS cells promote organoid formation of single Lgr5(+) colon stem cells. DCS cells can be massively produced from Lgr5(+) colon stem cells in vitro by combined Notch inhibition and Wnt activation. We conclude that Reg4(+) DCS cells serve as Paneth cell equivalents in the colon crypt niche.

KW - intestinal stem cell

KW - niche

KW - Lgr5

KW - Reg4

KW - deep crypt secretory cells

U2 - 10.1073/pnas.1607327113

DO - 10.1073/pnas.1607327113

M3 - Article

C2 - 27573849

SN - 0027-8424

VL - 113

SP - E5399-E5407

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 37

ER -