A distinct role for Lgr5+ stem cells in primary and metastatic colon cancer

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Cancer stem cells (CSCs) have been hypothesized to represent the driving force behind tumour progression and metastasis, making them attractive cancer targets. However, conclusive experimental evidence for their functional relevance is still lacking for most malignancies. Here we show that the leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5) identifies intestinal CSCs in mouse tumours engineered to recapitulate the clinical progression of human colorectal cancer. We demonstrate that selective Lgr5+ cell ablation restricts primary tumour growth, but does not result in tumour regression. Instead, tumours are maintained by proliferative Lgr5 cells that continuously attempt to replenish the Lgr5+ CSC pool, leading to rapid re-initiation of tumour growth upon treatment cessation. Notably, CSCs are critical for the formation and maintenance of liver metastasis derived from colorectal cancers. Together, our data highlight distinct CSC dependencies for primary versus metastasic tumour growth, and suggest that targeting CSCs may represent a therapeutic opportunity for managing metastatic disease.

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We thank M. Junttila and K. Leong for generation of the AKVL mouse model, A. Bruce for help designing schematic figures, J. Jian for tissue processing, J. Stinson and the NGS laboratory for RNA-sequencing, J. Paw, C. Poon, T. Ho and J. Borneo for flow cytometry support, members of Laboratory Animal Resources for animal care and de Sauvage laboratory members for comments on the manuscript.

Author information


  1. Molecular Oncology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA

    • Felipe de Sousa e Melo
    • , Antonina V. Kurtova
    • , Noelyn Kljavin
    • , Joerg D. Hoeck
    • , Elaine E. Storm
    • , Gerrit J. P. Dijkgraaf
    •  & Frederic J. de Sauvage
  2. Cancer Immunology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA

    • Jonathan M. Harnoss
  3. Research Pathology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA

    • Jeffrey Hung
    • , Jeffrey Eastham Anderson
    •  & Hartmut Koeppen
  4. Molecular Biology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA

    • Zora Modrusan
  5. Bioinformatics and Computational Biology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA

    • Robert Piskol


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F.d.S.M. and F.J.d.S. conceptualized the project and designed experiments. F.d.S.M. engineered and characterized all mutant organoid lines, produced concentrated lentivirus, optimized the orthotopic injection methodology and conducted bioluminescent imaging. F.d.S.M., A.V.K. and N.K. performed subcutaneous transplants, endoscopic imaging, DT treatment, tumour measurements and collected tissues. F.d.S.M. and A.V.K. conducted flow cytometry experiments. J.M.H. performed portal vein injections. G.J.P.D. modified pLKO-SHC201, cloned sgRNAs and designed pGEIL-tRFP. F.d.S.M., J.D.H., J.H., J.E.A. and E.E.S. either acquired or evaluated immunohistochemistry and/or immunofluorescence data. H.K. performed histopathological evaluation of tumour and liver tissues. Z.M. contributed next-generation sequencing data. R.P. provided bioinformatics support. F.d.S.M., G.J.P.D. and F.J.d.S. wrote the manuscript.

Competing interests

All authors are employees of Genentech and own Roche shares.

Corresponding author

Correspondence to Frederic J. de Sauvage.

Reviewer Information Nature thanks F. Greten, R. Shivdasani and the 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

Excel files

  1. 1.

    Supplementary Table 1

    Off-target prediction for Trp53 and Smad4 sgRNAs.

  2. 2.

    Supplementary Table 2

    Time course analysis of transcriptional changes after DT treatment in vivo.

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    Supplementary Table 3

    Quantitative gene set analysis of expression changes in tumours after DT treatment.

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    Supplementary Table 4

    Quantitative gene set analysis of expression changes of selected gene sets.


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