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FOXO1 inhibition yields functional insulin-producing cells in human gut organoid cultures

Nature Communications volume 5, Article number: 4242 (2014) | Download Citation

Abstract

Generation of surrogate sources of insulin-producing β-cells remains a goal of diabetes therapy. While most efforts have been directed at differentiating embryonic or induced pluripotent stem (iPS) cells into β-like-cells through endodermal progenitors, we have shown that gut endocrine progenitor cells of mice can be differentiated into glucose-responsive, insulin-producing cells by ablation of transcription factor Foxo1. Here we show that FOXO1 is present in human gut endocrine progenitor and serotonin-producing cells. Using gut organoids derived from human iPS cells, we show that FOXO1 inhibition using a dominant-negative mutant or lentivirus-encoded small hairpin RNA promotes generation of insulin-positive cells that express all markers of mature pancreatic β-cells, release C-peptide in response to secretagogues and survive in vivo following transplantation into mice. The findings raise the possibility of using gut-targeted FOXO1 inhibition or gut organoids as a source of insulin-producing cells to treat human diabetes.

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References

  1. 1.

    et al. A scalable system for production of functional pancreatic progenitors from human embryonic stem cells. PLoS ONE 7, e37004 (2012).

  2. 2.

    et al. Functional beta-cell maturation is marked by an increased glucose threshold and by expression of urocortin 3. Nat. Biotechnol. 30, 261–264 (2012).

  3. 3.

    et al. iPSC-derived beta cells model diabetes due to glucokinase deficiency. J. Clin. Invest. 123, 3146–3153 (2013).

  4. 4.

    et al. Generation of pluripotent stem cells from patients with type 1 diabetes. Proc. Natl Acad. Sci. USA 106, 15768–15773 (2009).

  5. 5.

    , & Minireview: development and differentiation of gut endocrine cells. Endocrinology 145, 2639–2644 (2004).

  6. 6.

    , , , & Generation of functional insulin-producing cells in the gut by Foxo1 ablation. Nat. Genet. 44, 406–412 (2012).

  7. 7.

    et al. Overlap of endocrine hormone expression in the mouse intestine revealed by transcriptional profiling and flow cytometry. Endocrinology 153, 3054–3065 (2012).

  8. 8.

    , , , & Pancreatic beta cell dedifferentiation as a mechanism of diabetic beta cell failure. Cell 150, 1223–1234 (2012).

  9. 9.

    et al. Conversion of adult pancreatic alpha-cells to beta-cells after extreme beta-cell loss. Nature 464, 1149–1154 (2010).

  10. 10.

    et al. Beta cells can be generated from endogenous progenitors in injured adult mouse pancreas. Cell 132, 197–207 (2008).

  11. 11.

    & FoxOs at the crossroads of cellular metabolism, differentiation, and transformation. Cell 117, 421–426 (2004).

  12. 12.

    et al. Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro. Nature 470, 105–109 (2011).

  13. 13.

    , , , & OLFM4 is a robust marker for stem cells in human intestine and marks a subset of colorectal cancer cells. Gastroenterology 137, 15–17 (2009).

  14. 14.

    et al. EphB3 marks delaminating endocrine progenitor cells in the developing pancreas. Dev. Dyn. 241, 1008–1019 (2012).

  15. 15.

    , , & Current view: intestinal stem cells and signaling. Gastroenterology 134, 849–864 (2008).

  16. 16.

    , , & Neurogenin3 is required for the development of the four endocrine cell lineages of the pancreas. Proc. Natl Acad. Sci. USA 97, 1607–1611 (2000).

  17. 17.

    et al. Expression of neurogenin3 reveals an islet cell precursor population in the pancreas. Development 127, 3533–3542 (2000).

  18. 18.

    , , & Neurogenin 3 is essential for the proper specification of gastric enteroendocrine cells and the maintenance of gastric epithelial cell identity. Genes Dev. 16, 1488–1497 (2002).

  19. 19.

    et al. Convergence of the insulin and serotonin programs in the pancreatic beta-cell. Diabetes 60, 3208–3216 (2011).

  20. 20.

    , , , & Expression of PCSK1 (PC1/3), PCSK2 (PC2) and PCSK3 (furin) in mouse small intestine. Regul. Pept. 152, 54–60 (2009).

  21. 21.

    , , , & Differential processing of pro-glucose-dependent insulinotropic polypeptide in gut. Am. J. Physiol. Gastrointest. Liver Physiol. 298, G608–G614 (2010).

  22. 22.

    et al. Co-localisation of the Kir6.2/SUR1 channel complex with glucagon-like peptide-1 and glucose-dependent insulinotrophic polypeptide expression in human ileal cells and implications for glycaemic control in new onset type 1 diabetes. Eur. J. Endocrinol. 156, 663–671 (2007).

  23. 23.

    , , & Generating human intestinal tissue from pluripotent stem cells in vitro. Nat. Protoc. 6, 1920–1928 (2011).

  24. 24.

    , , & The forkhead transcription factor Foxo1 (Fkhr) confers insulin sensitivity onto glucose-6-phosphatase expression. J. Clin. Invest. 108, 1359–1367 (2001).

  25. 25.

    et al. A Foxo/Notch pathway controls myogenic differentiation and fiber type specification. J. Clin. Invest. 117, 2477–2485 (2007).

  26. 26.

    et al. Glucose-dependent insulin release from genetically engineered K cells. Science 290, 1959–1962 (2000).

  27. 27.

    et al. Serotonin regulates glucose-stimulated insulin secretion from pancreatic beta cells during pregnancy. Proc. Natl Acad. Sci. USA 110, 19420–19425 (2013).

  28. 28.

    et al. De novo formation of insulin-producing ‘neo-beta cell islets’ from intestinal crypts. Cell Rep. 6, 1046–1058 (2014).

  29. 29.

    et al. Inhibition of Notch signaling ameliorates insulin resistance in a FoxO1-dependent manner. Nat. Med. 17, 961–967 (2011).

  30. 30.

    et al. The forkhead transcription factor Foxo1 links insulin signaling to Pdx1 regulation of pancreatic beta cell growth. J. Clin. Invest. 110, 1839–1847 (2002).

  31. 31.

    et al. Effects of the novel Foxo1 inhibitor AS1708727 on plasma glucose and triglyceride levels in diabetic db/db mice. Eur. J. Pharmacol. 645, 185–191 (2010).

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Acknowledgements

R.B. is supported by a fellowship from the Manpei Suzuki Diabetes Foundation. K.S.F. was supported by a fellowship from the Swedish Society for Medical Research. K.T. was supported by the Japan Society for the Promotion of Science. This work was supported by NIH grants DK58282 and DK63608, by a collaborative research agreement with Astra-Zeneca Corporation, by the JPB Foundation, by the New York Stem Cell Foundation and by the Brehm Coalition. We thank members of the Accili and Leibel laboratories for discussion of the data and critical reading of the manuscript. We thank A. Stewart and N. Fiaschi-Taesch (Mount Sinai School of Medicine, NY) for gifting human islets.

Author information

Affiliations

  1. Department of Medicine, Columbia University College of Physicians and Surgeons, New York, New York 10032, USA

    • Ryotaro Bouchi
    •  & Domenico Accili
  2. New York Stem Cell Foundation Research Institute, New York, New York 10032, USA

    • Kylie S. Foo
    • , Haiqing Hua
    •  & Dieter Egli
  3. Department of Pediatrics, Columbia University College of Physicians and Surgeons, New York, New York 10032, USA

    • Kylie S. Foo
    • , Haiqing Hua
    • , Dieter Egli
    •  & Rudolph L. Leibel
  4. Tokyo Medical and Dental University, Tokyo 113-8510, Japan

    • Kyoichiro Tsuchiya
  5. Department of Surgery, Columbia University College of Physicians and Surgeons, New York, New York 10032, USA

    • Yoshiaki Ohmura
    • , P. Rodrigo Sandoval
    •  & Lloyd E. Ratner

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Contributions

R.B. designed and performed experiments, analysed data and wrote the manuscript. K.S.F., H.H. and K.T. designed and performed experiments. P.R.S., Y.O. and L.E.R. procured surgical specimens; D.E., R.L.L. and D.A. designed experiments, oversaw research and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Domenico Accili.

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https://doi.org/10.1038/ncomms5242

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