Most patients with colorectal cancer die as a result of the disease spreading to other organs. However, no prevalent mutations have been associated with metastatic colorectal cancers1,2. Instead, particular features of the tumour microenvironment, such as lack of T-cell infiltration3, low type 1 T-helper cell (TH1) activity and reduced immune cytotoxicity2 or increased TGFβ levels4 predict adverse outcomes in patients with colorectal cancer. Here we analyse the interplay between genetic alterations and the tumour microenvironment by crossing mice bearing conditional alleles of four main colorectal cancer mutations in intestinal stem cells. Quadruple-mutant mice developed metastatic intestinal tumours that display key hallmarks of human microsatellite-stable colorectal cancers, including low mutational burden5, T-cell exclusion3 and TGFβ-activated stroma4,6,7. Inhibition of the PD-1–PD-L1 immune checkpoint provoked a limited response in this model system. By contrast, inhibition of TGFβ unleashed a potent and enduring cytotoxic T-cell response against tumour cells that prevented metastasis. In mice with progressive liver metastatic disease, blockade of TGFβ signalling rendered tumours susceptible to anti-PD-1–PD-L1 therapy. Our data show that increased TGFβ in the tumour microenvironment represents a primary mechanism of immune evasion that promotes T-cell exclusion and blocks acquisition of the TH1-effector phenotype. Immunotherapies directed against TGFβ signalling may therefore have broad applications in treating patients with advanced colorectal cancer.

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Change history

  • Corrected online 15 February 2018

    An incorrect present address was listed for Adrià Cañellas; this has now been corrected online.


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  1. 1.

    et al. Comparative lesion sequencing provides insights into tumor evolution. Proc. Natl Acad. Sci. USA 105, 4283–4288 (2008)

  2. 2.

    et al. The tumor microenvironment and immunoscore are critical determinants of dissemination to distant metastasis. Sci. Transl. Med. 8, 327ra26 (2016)

  3. 3.

    et al. Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science 313, 1960–1964 (2006)

  4. 4.

    et al. Stromal gene expression defines poor-prognosis subtypes in colorectal cancer. Nat. Genet. 47, 320–329 (2015)

  5. 5.

    Cancer Genome Atlas Network. Comprehensive molecular characterization of human colon and rectal cancer. Nature 487, 330–337 (2012)

  6. 6.

    et al. Dependency of colorectal cancer on a TGF-β-driven program in stromal cells for metastasis initiation. Cancer Cell 22, 571–584 (2012)

  7. 7.

    et al. The consensus molecular subtypes of colorectal cancer. Nat. Med. 21, 1350–1356 (2015)

  8. 8.

    & A genetic model for colorectal tumorigenesis. Cell 61, 759–767 (1990)

  9. 9.

    et al. Genetic dissection of colorectal cancer progression by orthotopic transplantation of engineered cancer organoids. Proc. Natl Acad. Sci. USA 114, E2357–E2364 (2017)

  10. 10.

    et al. Transplantation of engineered organoids enables rapid generation of metastatic mouse models of colorectal cancer. Nat. Biotechnol. 35, 577–582 (2017)

  11. 11.

    et al. A distinct role for Lgr5+ stem cells in primary and metastatic colon cancer. Nature 543, 676–680 (2017)

  12. 12.

    et al. Liver-targeted disruption of Apc in mice activates β-catenin signaling and leads to hepatocellular carcinomas. Proc. Natl Acad. Sci. USA 101, 17216–17221 (2004)

  13. 13.

    et al. Analysis of lung tumor initiation and progression using conditional expression of oncogenic K-ras. Genes Dev. 15, 3243–3248 (2001)

  14. 14.

    , , , & Induction of medulloblastomas in p53-null mutant mice by somatic inactivation of Rb in the external granular layer cells of the cerebellum. Genes Dev. 14, 994–1004 (2000)

  15. 15.

    et al. Induced disruption of the transforming growth factor beta type II receptor gene in mice causes a lethal inflammatory disorder that is transplantable. Blood 100, 560–568 (2002)

  16. 16.

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

  17. 17.

    et al. Stromal contribution to the colorectal cancer transcriptome. Nat. Genet. 47, 312–319 (2015)

  18. 18.

    et al. Prospective derivation of a living organoid biobank of colorectal cancer patients. Cell 161, 933–945 (2015)

  19. 19.

    et al. A colorectal tumor organoid library demonstrates progressive loss of niche factor requirements during tumorigenesis. Cell Stem Cell 18, 827–838 (2016)

  20. 20.

    et al. Mutational heterogeneity in cancer and the search for new cancer-associated genes. Nature 499, 214–218 (2013)

  21. 21.

    et al. Signatures of mutational processes in human cancer. Nature 500, 415–421 (2013)

  22. 22.

    et al. First-in-human dose study of the novel transforming growth factor-β receptor I kinase inhibitor LY2157299 monohydrate in patients with advanced cancer and glioma. Clin. Cancer Res. 21, 553–560 (2015)

  23. 23.

    et al. The vigorous immune microenvironment of microsatellite instable colon cancer is balanced by multiple counter-inhibitory checkpoints. Cancer Discov. 5, 43–51 (2015)

  24. 24.

    et al. PD-1 blockade in tumors with mismatch-repair deficiency. N. Engl. J. Med. 372, 2509–2520 (2015)

  25. 25.

    & Abrogation of TGFβ signaling in T cells leads to spontaneous T cell differentiation and autoimmune disease. Immunity 12, 171–181 (2000)

  26. 26.

    , & Cellular mechanisms of fatal early-onset autoimmunity in mice with the T cell-specific targeting of transforming growth factor-β receptor. Immunity 25, 441–454 (2006)

  27. 27.

    & TGF-β directly targets cytotoxic T cell functions during tumor evasion of immune surveillance. Cancer Cell 8, 369–380 (2005)

  28. 28.

    , , & TGF-β signaling regulates CD8+ T cell responses to high- and low-affinity TCR interactions. Int. Immunol. 17, 531–538 (2005)

  29. 29.

    & Elements of cancer immunity and the cancer-immune set point. Nature 541, 321–330 (2017)

  30. 30.

    & T cell exclusion, immune privilege, and the tumor microenvironment. Science 348, 74–80 (2015)

  31. 31.

    et al. Deletion of the developmentally essential gene Atr in adult mice leads to age-related phenotypes and stem cell loss. Cell Stem Cell 1, 113–126 (2007)

  32. 32.

    , , , & A global double-fluorescent Cre reporter mouse. Genesis 45, 593–605 (2007)

  33. 33.

    R Core Team. R: a Language and Environment for Statistical Computing (R Foundation for Statistical Computing, 2017)

  34. 34.

    ggplot2: Elegant Graphics for Data Analysis (Springer, 2009)

  35. 35.

    , , , & Regulation by vascular endothelial growth factor of human colon cancer tumorigenesis in a mouse model of experimental liver metastasis. J. Clin. Invest. 95, 1789–1797 (1995)

  36. 36.

    et al. Orthotopic microinjection of human colon cancer cells in nude mice induces tumor foci in all clinically relevant metastatic sites. Am. J. Pathol. 170, 1077–1085 (2007)

  37. 37.

    , , & A Lego system for conditional inference. Am. Stat. 60, 257–263 (2006)

  38. 38.

    & Data analysis using regression and multilevel/hierarchical models. J. Stat. Softw. 30, 1–5 (2009)

  39. 39.

    , , & Fitting linear mixed-effects models using lme4. J. Stat. Softw. 67, 1–48 (2015)

  40. 40.

    , & B. lmerTest Package: tests in linear mixed effects models J. Stat. Softw. 82, 1–26 (2015)

  41. 41.

    et al. Immune response in silico (IRIS): immune-specific genes identified from a compendium of microarray expression data. Genes Immun. 6, 319–331 (2005)

  42. 42.

    & Controlling the false discovery rate: a practical and powerful approach to multiple testing. J. R. Stat. Soc. B 57, 289–300 (1995)

  43. 43.

    , , , & (eds) Bioinformatics and Computational Biology Solutions using R and Bioconductor. (Springer, 2005)

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We thank E. Sancho for critical reading of this manuscript, all members of the Batlle laboratory for support and discussions, and members of the López-Bigas laboratory for help with revising the manuscript. We are grateful for the assistance of the IRB Barcelona core facilities for histopathology, functional genomics, mouse mutant and advanced digital microscopy; the flow cytometry and animal facilities of the UB/PCB; and the CRG genomic unit. D.V.F.T. held a Juan de la Cierva postdoctoral fellowship from MINECO. This work was supported by grants from the Doctor Josef Steiner Foundation, ERC advanced grant 340176, Instituto de Salud Carlos III, Olga Torres Foundation, BBVA Foundation, grant SAF-2014-53784 (MINECO) and by Fundación Botín. IRB Barcelona is the recipient of a Severo Ochoa Award of Excellence from MINECO.

Author information

Author notes

    • Alexandre Calon
    •  & Elisa I. Rivas

    Present address: Cancer Research Programme, Hospital del Mar Research Institute (IMIM), 08003 Barcelona, Spain (A.C. and E.I.R.).


  1. Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Baldiri i Reixac 10, 08028 Barcelona, Spain

    • Daniele V. F. Tauriello
    • , Sergio Palomo-Ponce
    • , Diana Stork
    • , Antonio Berenguer-Llergo
    • , Jordi Badia-Ramentol
    • , Marta Sevillano
    • , Sales Ibiza
    • , Adrià Cañellas
    • , Xavier Hernando-Momblona
    • , Daniel Byrom
    • , Joan A. Matarin
    • , Alexandre Calon
    • , Elisa I. Rivas
    • , Angel R. Nebreda
    • , Antoni Riera
    • , Camille Stephan-Otto Attolini
    •  & Eduard Batlle
  2. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Barcelona, Spain

    • Daniele V. F. Tauriello
    • , Sergio Palomo-Ponce
    • , Mar Iglesias
    • , Marta Sevillano
    • , Xavier Hernando-Momblona
    •  & Eduard Batlle
  3. Department of Pathology, Hospital del Mar, 08003 Barcelona, Spain

    • Mar Iglesias
  4. Cancer Research Programme, Hospital del Mar Research Institute (IMIM), 08003 Barcelona, Spain

    • Mar Iglesias
  5. Autonomous University of Barcelona (UAB), Spain

    • Mar Iglesias
  6. ICREA, Passeig Lluís Companys 23, 08010 Barcelona, Spain

    • Angel R. Nebreda
    •  & Eduard Batlle
  7. Department of Organic Chemistry, University of Barcelona, Martí i Franqués 1-11, 08028 Barcelona, Spain

    • Antoni Riera


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D.V.F.T., S.P.-P., D.S. and A.Cal. performed animal husbandry and genotyping; M.S. performed immunohistochemistry; D.V.F.T. and M.I. analysed histopathology. D.V.F.T. generated MTOs, which D.V.F.T. and D.S. characterized in vitro; D.S. performed CRISPR experiments; C.S.-O.A. and A.B.-L. performed exome and RNA-seq analyses, other bioinformatics (CMS classifier and patient data), and statistical analyses. S.P.-P., D.V.F.T., J.B.-R., A.Cañ. and X.H.-M. performed mouse isografting; D.V.F.T., D.S. and J.B.-R. quantified immunohistochemistry. D.B., J.A.M. and A.R. synthesized galunisertib. D.V.F.T. coordinated and performed animal treatments and analysed the data. D.V.F.T., J.B.-R., S.I., E.I.R. and A.R.N. performed immunophenotyping experiments. E.B. and D.V.F.T. conceived the study, coordinated experiments and wrote the manuscript. E.B. supervised the study.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Eduard Batlle.

Reviewer Information Nature thanks L. Vermeulen and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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    This file contains the uncropped western blots, a Supplementary Discussion, Supplementary Acknowledgements, Supplementary Methods and Supplementary References

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