The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers

Javier Muñoz; Daniel E. Stange; Arnout G. Schepers; Marc van de Wetering; Bon-Kyoung Koo; Shalev Itzkovitz; Richard Volckmann; Kevin S. Kung; Jan Koster; Sorina Radulescu; Kevin Myant; Rogier Versteeg; Owen J. Sansom; Johan H. van Es; Nick Barker; Alexander van Oudenaarden; Shabaz Mohammed; Albert J. R. Heck; Hans Clevers

doi:https://doi.org/10.1038/emboj.2012.166

The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers

Javier Muñoz, Daniel E. Stange, Arnout G. Schepers, Marc van de Wetering, Bon-Kyoung Koo, Shalev Itzkovitz, Richard Volckmann, Kevin S. Kung, Jan Koster, Sorina Radulescu, Kevin Myant, Rogier Versteeg, Owen J. Sansom, Johan H. van Es, Nick Barker, Alexander van Oudenaarden, Shabaz Mohammed, Albert J. R. Heck, Hans Clevers

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

573 Citations (Scopus)

Abstract

Two types of stem cells are currently defined in small intestinal crypts: cycling crypt base columnar (CBC) cells and quiescent '+4' cells. Here, we combine transcriptomics with proteomics to define a definitive molecular signature for Lgr5(+) CBC cells. Transcriptional profiling of FACS-sorted Lgr5(+) stem cells and their daughters using two microarray platforms revealed an mRNA stem cell signature of 384 unique genes. Quantitative mass spectrometry on the same cell populations identified 278 proteins enriched in intestinal stem cells. The mRNA and protein data sets showed a high level of correlation and a combined signature of 510 stem cell-enriched genes was defined. Spatial expression patterns were further characterized by mRNA in-situ hybridization, revealing that approximately half of the genes were expressed in a gradient with highest levels at the crypt bottom, while the other half was expressed uniquely in Lgr5(+) stem cells. Lineage tracing using a newly established knock-in mouse for one of the signature genes, Smoc2, confirmed its stem cell specificity. Using this resource, we find-and confirm by independent approaches-that the proposed quiescent/'+4' stem cell markers Bmi1, Tert, Hopx and Lrig1 are robustly expressed in CBC cells. The EMBO Journal (2012) 31, 3079-3091. doi:10.1038/emboj.2012.166; Published online 12 June 2012

Original language	English
Pages (from-to)	3079-3091
Journal	EMBO Journal
Volume	31
Issue number	14
DOIs	https://doi.org/10.1038/emboj.2012.166
Publication status	Published - 2012

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https://doi.org/10.1038/emboj.2012.166

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Muñoz, J., Stange, D. E., Schepers, A. G., van de Wetering, M., Koo, B.-K., Itzkovitz, S., Volckmann, R., Kung, K. S., Koster, J., Radulescu, S., Myant, K., Versteeg, R., Sansom, O. J., van Es, J. H., Barker, N., van Oudenaarden, A., Mohammed, S., Heck, A. J. R., & Clevers, H. (2012). The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers. EMBO Journal, 31(14), 3079-3091. https://doi.org/10.1038/emboj.2012.166

@article{48be8b6706e945e9940cfd966f0071de,

title = "The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers",

abstract = "Two types of stem cells are currently defined in small intestinal crypts: cycling crypt base columnar (CBC) cells and quiescent '+4' cells. Here, we combine transcriptomics with proteomics to define a definitive molecular signature for Lgr5(+) CBC cells. Transcriptional profiling of FACS-sorted Lgr5(+) stem cells and their daughters using two microarray platforms revealed an mRNA stem cell signature of 384 unique genes. Quantitative mass spectrometry on the same cell populations identified 278 proteins enriched in intestinal stem cells. The mRNA and protein data sets showed a high level of correlation and a combined signature of 510 stem cell-enriched genes was defined. Spatial expression patterns were further characterized by mRNA in-situ hybridization, revealing that approximately half of the genes were expressed in a gradient with highest levels at the crypt bottom, while the other half was expressed uniquely in Lgr5(+) stem cells. Lineage tracing using a newly established knock-in mouse for one of the signature genes, Smoc2, confirmed its stem cell specificity. Using this resource, we find-and confirm by independent approaches-that the proposed quiescent/'+4' stem cell markers Bmi1, Tert, Hopx and Lrig1 are robustly expressed in CBC cells. The EMBO Journal (2012) 31, 3079-3091. doi:10.1038/emboj.2012.166; Published online 12 June 2012",

author = "Javier Mu{\~n}oz and Stange, {Daniel E.} and Schepers, {Arnout G.} and {van de Wetering}, Marc and Bon-Kyoung Koo and Shalev Itzkovitz and Richard Volckmann and Kung, {Kevin S.} and Jan Koster and Sorina Radulescu and Kevin Myant and Rogier Versteeg and Sansom, {Owen J.} and {van Es}, {Johan H.} and Nick Barker and {van Oudenaarden}, Alexander and Shabaz Mohammed and Heck, {Albert J. R.} and Hans Clevers",

year = "2012",

doi = "https://doi.org/10.1038/emboj.2012.166",

language = "English",

volume = "31",

pages = "3079--3091",

journal = "EMBO Journal",

issn = "0261-4189",

publisher = "Wiley-Blackwell",

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Muñoz, J, Stange, DE, Schepers, AG, van de Wetering, M, Koo, B-K, Itzkovitz, S, Volckmann, R, Kung, KS, Koster, J, Radulescu, S, Myant, K, Versteeg, R, Sansom, OJ, van Es, JH, Barker, N, van Oudenaarden, A, Mohammed, S, Heck, AJR & Clevers, H 2012, 'The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers', EMBO Journal, vol. 31, no. 14, pp. 3079-3091. https://doi.org/10.1038/emboj.2012.166

TY - JOUR

T1 - The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers

AU - Muñoz, Javier

AU - Stange, Daniel E.

AU - Schepers, Arnout G.

AU - van de Wetering, Marc

AU - Koo, Bon-Kyoung

AU - Itzkovitz, Shalev

AU - Volckmann, Richard

AU - Kung, Kevin S.

AU - Koster, Jan

AU - Radulescu, Sorina

AU - Myant, Kevin

AU - Versteeg, Rogier

AU - Sansom, Owen J.

AU - van Es, Johan H.

AU - Barker, Nick

AU - van Oudenaarden, Alexander

AU - Mohammed, Shabaz

AU - Heck, Albert J. R.

AU - Clevers, Hans

PY - 2012

Y1 - 2012

N2 - Two types of stem cells are currently defined in small intestinal crypts: cycling crypt base columnar (CBC) cells and quiescent '+4' cells. Here, we combine transcriptomics with proteomics to define a definitive molecular signature for Lgr5(+) CBC cells. Transcriptional profiling of FACS-sorted Lgr5(+) stem cells and their daughters using two microarray platforms revealed an mRNA stem cell signature of 384 unique genes. Quantitative mass spectrometry on the same cell populations identified 278 proteins enriched in intestinal stem cells. The mRNA and protein data sets showed a high level of correlation and a combined signature of 510 stem cell-enriched genes was defined. Spatial expression patterns were further characterized by mRNA in-situ hybridization, revealing that approximately half of the genes were expressed in a gradient with highest levels at the crypt bottom, while the other half was expressed uniquely in Lgr5(+) stem cells. Lineage tracing using a newly established knock-in mouse for one of the signature genes, Smoc2, confirmed its stem cell specificity. Using this resource, we find-and confirm by independent approaches-that the proposed quiescent/'+4' stem cell markers Bmi1, Tert, Hopx and Lrig1 are robustly expressed in CBC cells. The EMBO Journal (2012) 31, 3079-3091. doi:10.1038/emboj.2012.166; Published online 12 June 2012

AB - Two types of stem cells are currently defined in small intestinal crypts: cycling crypt base columnar (CBC) cells and quiescent '+4' cells. Here, we combine transcriptomics with proteomics to define a definitive molecular signature for Lgr5(+) CBC cells. Transcriptional profiling of FACS-sorted Lgr5(+) stem cells and their daughters using two microarray platforms revealed an mRNA stem cell signature of 384 unique genes. Quantitative mass spectrometry on the same cell populations identified 278 proteins enriched in intestinal stem cells. The mRNA and protein data sets showed a high level of correlation and a combined signature of 510 stem cell-enriched genes was defined. Spatial expression patterns were further characterized by mRNA in-situ hybridization, revealing that approximately half of the genes were expressed in a gradient with highest levels at the crypt bottom, while the other half was expressed uniquely in Lgr5(+) stem cells. Lineage tracing using a newly established knock-in mouse for one of the signature genes, Smoc2, confirmed its stem cell specificity. Using this resource, we find-and confirm by independent approaches-that the proposed quiescent/'+4' stem cell markers Bmi1, Tert, Hopx and Lrig1 are robustly expressed in CBC cells. The EMBO Journal (2012) 31, 3079-3091. doi:10.1038/emboj.2012.166; Published online 12 June 2012

U2 - https://doi.org/10.1038/emboj.2012.166

DO - https://doi.org/10.1038/emboj.2012.166

M3 - Article

C2 - 22692129

SN - 0261-4189

VL - 31

SP - 3079

EP - 3091

JO - EMBO Journal

JF - EMBO Journal

IS - 14

ER -