• A Corrigendum to this article was published on 10 April 2018

This article has been updated

Abstract

With the goal of modeling human disease of the large intestine, we sought to develop an effective protocol for deriving colonic organoids (COs) from differentiated human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs). Extensive gene and immunohistochemical profiling confirmed that the derived COs represent colon rather than small intestine, containing stem cells, transit-amplifying cells, and the expected spectrum of differentiated cells, including goblet and endocrine cells. We applied this strategy to iPSCs derived from patients with familial adenomatous polyposis (FAP-iPSCs) harboring germline mutations in the WNT-signaling-pathway-regulator gene encoding APC, and we generated COs that exhibit enhanced WNT activity and increased epithelial cell proliferation, which we used as a platform for drug testing. Two potential compounds, XAV939 and rapamycin, decreased proliferation in FAP-COs, but also affected cell proliferation in wild-type COs, which thus limits their therapeutic application. By contrast, we found that geneticin, a ribosome-binding antibiotic with translational 'read-through' activity, efficiently targeted abnormal WNT activity and restored normal proliferation specifically in APC-mutant FAP-COs. These studies provide an efficient strategy for deriving human COs, which can be used in disease modeling and drug discovery for colorectal disease.

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Change history

  • 13 March 2018

    In the version of this article initially published, there were several instances of duplication of confocal microscopy images presented in Figures 1e, 3i and 4c,h,j. These errors occurred during the final preparation of the figures, when representative images were selected to illustrate the results obtained from multiple independent experiments. To ensure that these errors did not affect the paper’s conclusions, the authors verified all raw data used to quantify the results. The results presented in the original Figures 3j and 4d were not affected, but the quantification in Figure 4i was. This quantification was performed again, and the results supported the original conclusions of the paper. In addition, the western blot shown in Figure 3d came from a different replicate experiment than the one represented in Supplementary Figure 9g,h (which shows the raw data). The original blot has been replaced with the correct blot that matches the raw data originally provided. The conclusions of this experiment remain unchanged. All affected images have been now corrected in the HTML and PDF versions of the article.

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Acknowledgements

S.C. is funded by The New York Stem Cell Foundation (R-103) and NIH/NIDDK (1 DP2 DK098093-01, DP3DK111907-01). This study was supported in part by a shared facility contract from the New York Department of Health (NYSTEM, C029156; to T.E. and S.C.); a Tri-institutional Stem Cell Initiative grant (2013-001; to F. M. and S.C.); and a pilot grant from the Center for Advanced Digestion Care (CADC) at Weill Cornell Medical College (to S.C. and S.L.). S.C. is New York Stem Cell Foundation-Robertson Investigator. This work was also supported by grants R03CA176788 (US National Institutes of Health/National Cancer Institute), the MD Anderson Cancer Center Institutional Research Grant (IRG) Program (E.V.), and a gift from the Feinberg Family to E.V.; Cancer Prevention Educational Award (R25T CA057730, US National Institutes of Health/National Cancer Institute (to K.C.); Arnold O. Beckman postdoctoral fellowship to H.J.C. We thank H.E. Varmus (Weill Cornell Medical College) for his support and J. Jin at Weill Cornell Medical College for kindly providing human aortic smooth muscle cells. We are grateful for the technical support and advice provided by H.S. Ralph in the Cell Screening Core Facility, J. McCormick in the Flow Cytometry Facility and L. Cohen-Gould in the Electron Microscopy Facility at Weill Cornell Medical College.

Author information

Author notes

    • Su-Yi Tsai

    Present address: Department of Life Sciences, National Taiwan University, Taipei, Taiwan.

Affiliations

  1. Department of Surgery, Weill Cornell Medical College, New York, New York, USA.

    • Miguel Crespo
    • , Su-Yi Tsai
    • , Sadaf Amin
    • , Miriam Gordillo
    • , Todd Evans
    •  & Shuibing Chen
  2. Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Eduardo Vilar
    •  & Kyle Chang
  3. Department of GI Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Eduardo Vilar
  4. Clinical Cancer Genetics Program, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Eduardo Vilar
  5. Weill Graduate School of Medical Sciences of Cornell University, New York, New York, USA.

    • Sadaf Amin
  6. Department of Medicine, Weill Cornell Medical College, New York, New York, USA.

    • Tara Srinivasan
    • , Mavee Witherspoon
    •  & Steven Lipkin
  7. Genomic Core, Weill Cornell Medical College, New York, New York, USA.

    • Tuo Zhang
    •  & Jenny Zhaoying Xiang
  8. Department of Biochemistry, Weill Cornell Medical College, New York, New York, USA.

    • Nina H Pipalia
    •  & Frederick R Maxfield
  9. Meyer Cancer Center, Weill Cornell Medical College, New York, New York, USA.

    • Huanhuan Joyce Chen
    •  & Shuibing Chen

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Contributions

S.C., T.E., S.L., and F.R.M. designed the project; M.C., S.-Y.T., S.A., T.S., N.H.P.H.J.C., M.W., and M.G. performed experiments; E.V., K.C., T. Z., and J.Z.X. performed the bioinformatics analysis; M.C., E.V., K.C., and S.C. analyzed data; and M.C., E.V., F.R.M., S.L., T.E., and S.C. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Todd Evans or Shuibing Chen.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Figures 1–9 and Supplementary Tables 1–3

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DOI

https://doi.org/10.1038/nm.4355

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