Letter | Published:

Interplay between metabolic identities in the intestinal crypt supports stem cell function

Nature volume 543, pages 424427 (16 March 2017) | Download Citation

Abstract

The small intestinal epithelium self-renews every four or five days. Intestinal stem cells (Lgr5+ crypt base columnar cells (CBCs)) sustain this renewal and reside between terminally differentiated Paneth cells at the bottom of the intestinal crypt1. Whereas the signalling requirements for maintaining stem cell function and crypt homeostasis have been well studied, little is known about how metabolism contributes to epithelial homeostasis. Here we show that freshly isolated Lgr5+ CBCs and Paneth cells from the mouse small intestine display different metabolic programs. Compared to Paneth cells, Lgr5+ CBCs display high mitochondrial activity. Inhibition of mitochondrial activity in Lgr5+ CBCs or inhibition of glycolysis in Paneth cells strongly affects stem cell function, as indicated by impaired organoid formation. In addition, Paneth cells support stem cell function by providing lactate to sustain the enhanced mitochondrial oxidative phosphorylation in the Lgr5+ CBCs. Mechanistically, we show that oxidative phosphorylation stimulates p38 MAPK activation by mitochondrial reactive oxygen species signalling, thereby establishing the mature crypt phenotype. Together, our results reveal a critical role for the metabolic identity of Lgr5+ CBCs and Paneth cells in supporting optimal stem cell function, and we identify mitochondria and reactive oxygen species signalling as a driving force of cellular differentiation.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    et al. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449, 1003–1007 (2007)

  2. 2.

    et al. UCP2 regulates energy metabolism and differentiation potential of human pluripotent stem cells. EMBO J. 30, 4860–4873 (2011)

  3. 3.

    & Reactive oxygen species prime Drosophila haematopoietic progenitors for differentiation. Nature 461, 537–541 (2009)

  4. 4.

    et al. Mitochondrial complex III ROS regulate adipocyte differentiation. Cell Metab. 14, 537–544 (2011)

  5. 5.

    et al. Mitochondrial reactive oxygen species promote epidermal differentiation and hair follicle development. Sci. Signal. 6, ra8 (2013)

  6. 6.

    et al. mTORC1 in the Paneth cell niche couples intestinal stem-cell function to calorie intake. Nature 486, 490–495 (2012)

  7. 7.

    et al. Engineering stem cell organoids. Cell Stem Cell 18, 25–38 (2016)

  8. 8.

    Adult intestinal stem cells: critical drivers of epithelial homeostasis and regeneration. Nat. Rev. Mol. Cell Biol. 15, 19–33 (2014)

  9. 9.

    et al. Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny. Nat. Methods 11, 106–112 (2014)

  10. 10.

    et al. Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. Nature 469, 415–418 (2011)

  11. 11.

    et al. Identification of Lgr5-independent spheroid-generating progenitors of the mouse fetal intestinal epithelium. Cell Reports 5, 421–432 (2013)

  12. 12.

    et al. Transplantation of expanded fetal intestinal progenitors contributes to colon regeneration after injury. Cell Stem Cell 13, 734–744 (2013)

  13. 13.

    & Growing self-organizing mini-guts from a single intestinal stem cell: mechanism and applications. Science 340, 1190–1194 (2013)

  14. 14.

    et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium. Gastroenterology 141, 1762–1772 (2011)

  15. 15.

    & Dichloroacetate and cancer: new home for an orphan drug? Biochim. Biophys. Acta 1846, 617–629 (2014)

  16. 16.

    , & Dichloroacetate (DCA) as a potential metabolic-targeting therapy for cancer. Br. J. Cancer 99, 989–994 (2008)

  17. 17.

    , , , & Availability of the key metabolic substrates dictates the respiratory response of cancer cells to the mitochondrial uncoupling. Biochim. Biophys. Acta 1837, 51–62 (2014)

  18. 18.

    et al. Galactose enhances oxidative metabolism and reveals mitochondrial dysfunction in human primary muscle cells. PLoS One 6, e28536 (2011)

  19. 19.

    et al. Real-time imaging of the intracellular glutathione redox potential. Nat. Methods 5, 553–559 (2008)

  20. 20.

    , , & p38 MAPK: a dual role in hepatocyte proliferation through reactive oxygen species. Free Radic. Res. 47, 905–916 (2013)

  21. 21.

    & Signal integration by JNK and p38 MAPK pathways in cancer development. Nat. Rev. Cancer 9, 537–549 (2009)

  22. 22.

    , , , & High-sensitivity measurements of multiple kinase activities in live single cells. Cell 157, 1724–1734 (2014)

  23. 23.

    et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459, 262–265 (2009)

  24. 24.

    et al. Secreted phospholipases A2 are intestinal stem cell niche factors with distinct roles in homeostasis, inflammation, and cancer. Cell Stem Cell 19, 38–51 (2016)

  25. 25.

    et al. The intestinal stem cell signature identifies colorectal cancer stem cells and predicts disease relapse. Cell Stem Cell 8, 511–524 (2011)

  26. 26.

    et al. Distinct effects of p38α deletion in myeloid lineage and gut epithelia in mouse models of inflammatory bowel disease. Gastroenterology 138, 1255–1265 (2010)

  27. 27.

    & Stem cells, redox signaling, and stem cell aging. Antioxid. Redox Signal. 20, 1902–1916 (2014)

  28. 28.

    , , , & Reactive oxygen species derived from the mitochondrial respiratory chain are not responsible for the basal levels of oxidative base modifications observed in nuclear DNA of Mammalian cells. Free Radic. Biol. Med. 36, 765–773 (2004)

  29. 29.

    et al. Mitochondrial reactive oxygen species are scavenged by Cockayne syndrome B protein in human fibroblasts without nuclear DNA damage. Proc. Natl Acad. Sci. USA 111, 13487–13492 (2014)

  30. 30.

    et al. FOXM1-induced PRX3 regulates stemness and survival of colon cancer cells via maintenance of mitochondrial function. Gastroenterology 149, 1006–1016 (2015)

  31. 31.

    , , , & A protocol for lentiviral transduction and downstream analysis of intestinal organoids. J. Vis. Exp. (2015)

  32. 32.

    et al. Synthetic combined superoxide dismutase/catalase mimetics are protective as a delayed treatment in a rat stroke model: a key role for reactive oxygen species in ischemic brain injury. J. Pharmacol. Exp. Ther. 284, 215–221 (1998)

  33. 33.

    , & Measuring EGSH and H2O2 with roGFP2-based redox probes. Free Radic. Biol. Med. 51, 1943–1951 (2011)

  34. 34.

    , & Listeria monocytogenes infection causes metabolic shifts in Drosophila melanogaster. PLoS One 7, e50679 (2012)

  35. 35.

    , , , & XCMS: processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching, and identification. Anal. Chem. 78, 779–787 (2006)

  36. 36.

    , , & MetaboAnalyst 3.0—making metabolomics more meaningful. Nucleic Acids Res. 43, W251–W257 (2015)

Download references

Acknowledgements

This work was financially supported by CGC.nl (M.J.R.-C., H.J.S.), Utrecht Life Sciences (M.M.), Dutch Cancer Society ((KWF), EMCR 2012-5473 (M.S.), and UU 2013-6070 (K.C.O.)) and from the Netherlands Institute of Regenerative Medicine (R.F.). We thank H. Bos, T. Dansen and S. van Mil for helpful discussions and proofreading; F. de Sauvage (Genentech) for providing DTR–LGR5–GFP mice; T. Dick (DKFZ) for mtGrx1–roGFP and I. Verlaan (UMC Utrecht) for R-spondin/Wnt3a-conditioned medium.

Author information

Affiliations

  1. Molecular Cancer Research, Center Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584CG Utrecht, The Netherlands

    • Maria J. Rodríguez-Colman
    • , Maaike Meerlo
    • , Edwin Stigter
    • , Marten Hornsveld
    • , Koen C. Oost
    • , Hugo J. Snippert
    •  & Boudewijn M. T. Burgering
  2. Department of Pathology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, The Netherlands

    • Matthias Schewe
    • , Andrea Sacchetti
    •  & Riccardo Fodde
  3. Department of Genetics and Center for Molecular Medicine, Lundlaan 6, 3584 EA Utrecht, The Netherlands

    • Johan Gerrits
    • , Mia Pras-Raves
    •  & Nanda Verhoeven-Duif

Authors

  1. Search for Maria J. Rodríguez-Colman in:

  2. Search for Matthias Schewe in:

  3. Search for Maaike Meerlo in:

  4. Search for Edwin Stigter in:

  5. Search for Johan Gerrits in:

  6. Search for Mia Pras-Raves in:

  7. Search for Andrea Sacchetti in:

  8. Search for Marten Hornsveld in:

  9. Search for Koen C. Oost in:

  10. Search for Hugo J. Snippert in:

  11. Search for Nanda Verhoeven-Duif in:

  12. Search for Riccardo Fodde in:

  13. Search for Boudewijn M. T. Burgering in:

Contributions

M.J.R.-C. and B.M.T.B. conceived the project, designed and performed experiments and wrote the manuscript; M.S. designed and performed organoid reconstitution experiments; M.M. performed experiments; E.S. and J.G. performed metabolic measurements. M.P.-R. and N.V.-D. performed metabolic data analysis. A.S. performed FACS of intestinal cells. M.H. performed p38 IHC. K.C.O. and H.J.S. provided organoid cultures, and H.J.S. co-wrote the manuscript. R.F. designed experiments and co-wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Boudewijn M. T. Burgering.

Reviewer Information Nature thanks T. Sato, A. Schulze and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Figure 1

    This file contains full scan images of western blots used for Extended Data 4a and b. Molecular weight markers are indicated; pp38 (phospho-p38) and p38. Dashed lines indicated cropped areas shown in Extended Data 4a and b.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature21673

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.