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Cited by 263Cancer Cells and MetastasisDigestive system and related healthWnt/β-catenin signaling in development and cancer

Identification of a cKit(+) colonic crypt base secretory cell that supports Lgr5(+) stem cells in mice

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Rothenberg, Michael E., Nusse, Ysbrand, Kalisky, Tomer, Lee, John J., Dalerba, Piero, Scheeren, Ferenc, Lobo, Neethan, Kulkarni, Subhash +6 more·2012·Gastroenterology
DOI 10.1053/j.gastro.2012.02.006PMC PMC3911891
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Abstract

Background & aims Paneth cells contribute to the small intestinal niche of Lgr5(+) stem cells. Although the colon also contains Lgr5(+) stem cells, it does not contain Paneth cells. We investigated the existence of colonic Paneth-like cells that have a distinct transcriptional signature and support Lgr5(+) stem cells. Methods We used multicolor fluorescence-activated cell sorting to isolate different subregions of colon crypts, based on known markers, from dissociated colonic epithelium of mice. We performed multiplexed single-cell gene expression analysis with quantitative reverse transcriptase polymerase chain reaction followed by hierarchical clustering analysis to characterize distinct cell types. We used immunostaining and fluorescence-activated cell sorting analyses with in vivo administration of a Notch inhibitor and in vitro organoid cultures to characterize different cell types. Results Multicolor fluorescence-activated cell sorting could isolate distinct regions of colonic crypts. Four major epithelial subtypes or transcriptional states were revealed by gene expression analysis of selected populations of single cells. One of these, the goblet cells, contained a distinct cKit/CD117(+) crypt base subpopulation that expressed Dll1, Dll4, and epidermal growth factor, similar to Paneth cells, which were also marked by cKit. In the colon, cKit(+) goblet cells were interdigitated with Lgr5(+) stem cells. In vivo, this colonic cKit(+) population was regulated by Notch signaling; administration of a γ-secretase inhibitor to mice increased the number of cKit(+) cells. When isolated from mouse colon, cKit(+) cells promoted formation of organoids from Lgr5(+) stem cells, which expressed Kitl/stem cell factor, the ligand for cKit. When organoids were depleted of cKit(+) cells using a toxin-conjugated antibody, organoid formation decreased. Conclusions cKit marks small intestinal Paneth cells and a subset of colonic goblet cells that are regulated by Notch signaling and support Lgr5(+) stem cells.

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  • Residual iKAP tumor cells shared gene expression signatures with intestinal Paneth cells but differed from Paneth-like cells in the colon and Reg4+ deep crypt secretory cells.

    “Comparative analysis revealed that residual tumor cells shared gene expression signatures with intestinal Paneth cells but differed from Paneth-like cells in the colon35(Figure S1G), as well as Reg4+ deep crypt secretory cells (Figure S1H), which are functionally analogous to Paneth cells in the murine colon.36,37”

    From Paneth-like transition drives resistance to dual targeting of KRAS and EGFR in colorectal cancer (2026) · cites this paper as [37]

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Authors
Rothenberg, M. E., Nusse, Y., Kalisky, T., Lee, J. J., Dalerba, P., Scheeren, F., Lobo, N., Kulkarni, S., Sim, S., Qian, D., Beachy, P. A., Pasricha, P. J., Quake, S. R., & Clarke, M. F.
Affiliations
  • Institute for Stem Cell Biology and Regenerative Medicine (US)
  • Stanford Medicine (US)
  • Howard Hughes Medical Institute (US)
Corresponding author
Clarke, Michael F.
ORCID
Venue
Gastroenterology
Publisher
Elsevier BV
Published
2012-05
Volume
142
Issue
5
Pages
1195-1205.e6
ISSN
0016-5085 (ISSN-L)
DOI
10.1053/j.gastro.2012.02.006
PMC
PMC3911891
OpenAlex
W2106173591
Cited by
263
References
5
Citations by year
2026: 4 · 2025: 7 · 2024: 9 · 2023: 23 · 2022: 20 · 2021: 21 · 2020: 21 · 2019: 24 · 2018: 25 · 2017: 23 · 2016: 20 · 2015: 18 · 2014: 13 · 2013: 21 · 2012: 13
FWCI
10.29
Type
journal article
Language
en
Open access
Licence
Locations
  • Gastroenterology (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • California Institute for Regenerative Medicine
  • National Institutes of Health (5P01CA139490-03, 5P30DK056339, T32 DK0070560)
Topics
  • Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Digestive system and related health
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
Keywords
CD117, goblet cells, Lgr5+ stem cells, Paneth cells, Notch signaling, DLL1, DLL4, stem cell niche, colonic organoids, stem cell factor, Kitl, colonic epithelium
MeSH terms
Animals, Cells, Cultured, Colon, Flow Cytometry, Mice, Inbred C57BL, Stem Cells, Antigens, CD, Cell Adhesion Molecules, Hyaluronan Receptors, Proto-Oncogene Proteins c-kit, Paneth Cells, Goblet Cells, Gene Expression Profiling, Receptors, G-Protein-Coupled, Mice, Receptors, Notch, Single-Cell Analysis

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