Abstract

Epithelial regeneration is critical for barrier maintenance and organ function after intestinal injury. The intestinal stem cell (ISC) niche provides Wnt, Notch and epidermal growth factor (EGF) signals supporting Lgr5+ crypt base columnar ISCs for normal epithelial maintenance1,2. However, little is known about the regulation of the ISC compartment after tissue damage. Using ex vivo organoid cultures, here we show that innate lymphoid cells (ILCs), potent producers of interleukin-22 (IL-22) after intestinal injury3,4, increase the growth of mouse small intestine organoids in an IL-22-dependent fashion. Recombinant IL-22 directly targeted ISCs, augmenting the growth of both mouse and human intestinal organoids, increasing proliferation and promoting ISC expansion. IL-22 induced STAT3 phosphorylation in Lgr5+ ISCs, and STAT3 was crucial for both organoid formation and IL-22-mediated regeneration. Treatment with IL-22 in vivo after mouse allogeneic bone marrow transplantation enhanced the recovery of ISCs, increased epithelial regeneration and reduced intestinal pathology and mortality from graft-versus-host disease. ATOH1-deficient organoid culture demonstrated that IL-22 induced epithelial regeneration independently of the Paneth cell niche. Our findings reveal a fundamental mechanism by which the immune system is able to support the intestinal epithelium, activating ISCs to promote regeneration.

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Acknowledgements

We gratefully acknowledge the technical assistance of the MSKCC Research Animal Resource Center and Molecular Cytology Core Facility. We also thank H. Clevers, H. Farin, S. Middendorp, C. Wiegerinck, J. van Es, M. van de Wetering, N. Sasaki, J. Sun and M. Li for their advice and critical evaluation of our work. This research was supported by National Institutes of Health award numbers K08-HL115355 (A.M.H.), R01-HL125571 (A.M.H.), R01-HL069929 (M.R.M.vdB.), R01-AI100288 (M.R.M.vdB.), R01-AI080455 (M.R.M.vdB.), R01-AI101406 (M.R.M.vdB.), P01-CA023766/Project 4 (R. J. O’Reilly/M.R.M.vdB.), K99-CA176376 (J.A.D.) and P30-CA008748 (MSKCC Core Grant). Support was also received from the US National Institute of Allergy and Infectious Diseases (NIAID contract HHSN272200900059C), the European Union (award GC220918, C. Blackburn), The Experimental Therapeutics Center of MSKCC funded by Mr William H. Goodwin and Mrs Alice Goodwin, The Lymphoma Foundation, Alex’s Lemonade Stand, The Geoffrey Beene Cancer Research Center at MSKCC, The Susan and Peter Solomon Divisional Genomics Program, MSKCC Cycle for Survival, and The Lucille Castori Center for Microbes, Inflammation & Cancer. T.C. was supported by Innovational Research Incentives Scheme Vidi grant 91710377 from the Netherlands Organization for Scientific Research (Zon-MW), and M.R.-H. was supported by the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme FP7/2007-2013 under REA grant agreement no. 289720. A.M.M. was supported by the Bio Medical Exchange Program of the Deutscher Akademischer Austauschdienst. C.A.L. was supported by Dutch Cancer Society clinical fellowship grant 2013-5883 and by a mobility grant from the University Medical Center Utrecht. J.A.D. was supported by a C. J. Martin fellowship from the Australian National Health and Medical Research Council, a Scholar Award from the American Society of Hematology, and the Mechtild Harf Research Grant from the DKMS Foundation for Giving Life. A.M.H. was supported by a Scholar Award from the American Society of Hematology, a New Investigator Award from the American Society for Blood and Marrow Transplantation, and the Amy Strelzer Manasevit Research Program. A provisional patent application has been filed on the use of IL-22 and F-652 as ISC growth factors (US 61/901,151) with A.M.H., C.A.L. and M.R.M.vdB. listed as inventors.

Author information

Author notes

    • Caroline A. Lindemans
    • , Marco Calafiore
    • , Anna M. Mertelsmann
    •  & Margaret H. O’Connor

    These authors contributed equally to this work.

    • Marcel R. M. van den Brink
    •  & Alan M. Hanash

    These authors jointly supervised this work.

Affiliations

  1. Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Caroline A. Lindemans
    • , Marco Calafiore
    • , Anna M. Mertelsmann
    • , Margaret H. O’Connor
    • , Robert R. Jenq
    • , Gillian Lawrence
    • , Juliet A. Ivanov
    • , Ya-Yuan Fu
    • , Shuichiro Takashima
    • , Marcel R. M. van den Brink
    •  & Alan M. Hanash
  2. Department of Pediatrics, University Medical Center Utrecht, 3508 AB Utrecht, The Netherlands

    • Caroline A. Lindemans
    • , Michal Mokry
    •  & Edward E. Nieuwenhuis
  3. Department of Immunology, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Jarrod A. Dudakov
    • , Enrico Velardi
    • , Lauren F. Young
    • , Odette M. Smith
    •  & Marcel R. M. van den Brink
  4. Department of Anatomy and Developmental Biology, Monash University, Clayton 3800, Australia

    • Jarrod A. Dudakov
  5. Department of Medicine, Weill Cornell Medicine, New York, New York 10021, USA

    • Robert R. Jenq
    •  & Lukas E. Dow
  6. Department of Radiation Oncology, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Guoqiang Hua
  7. Department of Molecular Pharmacology, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Guoqiang Hua
    • , Maria L. Martin
    •  & Richard Kolesnick
  8. Department of Cancer Biology & Genetics, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Kevin P. O’Rourke
  9. Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, USA

    • Yuan-Hung Lo
    •  & Noah F. Shroyer
  10. Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands

    • Monica Romera-Hernandez
    •  & Tom Cupedo
  11. Department of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, Florida 32610, USA

    • Chen Liu

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Contributions

C.A.L. and M.C. designed and performed organoid experiments. A.M.M. and M.H.O. performed and analysed in vivo experiments. J.A.D., R.R.J. and E.V. provided input and helped with various assays. L.F.Y., O.M.S. and G.L. performed and monitored bone marrow transplants and maintained the mouse colonies. J.A.I. assisted with organoid quantification. Y.-Y.F. analysed crypt sizes and confocal microscopy. S.T. assisted with ILC co-culture experiments. G.H., M.L.M. and R.K. assisted with ISC isolation and in vivo ISC quantification experiments and provided reagents and expertise. K.P.O. and L.D. assisted with adeno-Cre experiments and optimizing various assays. Y.-H.L. and N.F.S. assisted with Paneth cell deficiency experiments. M.M. and E.E.N. performed the GSEA analyses and assisted with reagents and resources. M.R.-H. performed PCR analyses on purified stem cells and immune cells under the guidance of T.C., and C.L. analysed intestinal histopathology. M.R.M.vdB. and A.M.H. supervised the research. All authors contributed to experimental design, interpretation and manuscript editing.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Alan M. Hanash.

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    Supplementary Information

    Supplementary Figure 1 shows the original uncropped images of western blots presented in Figures 2 and 4 and Supplementary Table 1 shows a list of antibodies used to perform flow cytometry, western blotting, and tissue staining for immunohistochemistry and immunofluorescence.

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https://doi.org/10.1038/nature16460

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