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 Born, Maaike Den
AU - Van Es, Johan H.
AU - Karthaus, Wouter R.
AU - Li, Vivian S.W.
AU - López-Iglesias, Carmen
AU - Peters, Peter J.
AU - Van Rheenen, Jacco
AU - Van Oudenaarden, Alexander
AU - Clevers, Hans
N1 - Funding Information:
We thank the Hubrecht mouse and imaging facility for taking care of mice and imaging analysis, E. R. Idris for his assistance during his internship course, Dr. Frederic J. de Sauvage (Genentech) for kindly providing us with Lgr5-DTR-eGFP mice, and the laboratory members for valuable discussions. This research was supported by grants to N. Sasaki [Japan Society for the Promotion of Science (JSPS) postdoctoral Fellowships for Research Abroad and Astellas Foundation for Research on Metabolic Disorders] and N. Sasaki, N. Sachs, H.B., M.v.d.B., J.H.v.E., W.R.K., V.S.W.L., and H.C. [National Institutes of Health (NIH)/Massachusetts Institute of Technology (MIT) Subaward 5710002735, Skolteck (SkTech) Skolokovo, and the European Union (EU) FP7 Tornado].
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 investigateanichefunction,weeliminatedDCScellsbyusingthediphtheriatoxin receptor gene knocked into the murine 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 investigateanichefunction,weeliminatedDCScellsbyusingthediphtheriatoxin receptor gene knocked into the murine 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 - Deep crypt secretory cells
KW - Intestinal stem cell
KW - Lgr5
KW - Niche
KW - Reg4
UR - http://www.scopus.com/inward/record.url?scp=85047289665&partnerID=8YFLogxK
U2 - 10.1073/pnas.1607327113
DO - 10.1073/pnas.1607327113
M3 - Article
C2 - 27573849
AN - SCOPUS:85047289665
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 -