Long-term persistence and development of induced pancreatic beta cells generated by lineage conversion of acinar cells

A Corrigendum to this article was published on 07 August 2015

This article has been updated

Abstract

Direct lineage conversion is a promising approach to generate therapeutically important cell types for disease modeling and tissue repair. However, the survival and function of lineage-reprogrammed cells in vivo over the long term has not been examined. Here, using an improved method for in vivo conversion of adult mouse pancreatic acinar cells toward beta cells, we show that induced beta cells persist for up to 13 months (the length of the experiment), form pancreatic islet–like structures and support normoglycemia in diabetic mice. Detailed molecular analyses of induced beta cells over 7 months reveal that global DNA methylation changes occur within 10 d, whereas the transcriptional network evolves over 2 months to resemble that of endogenous beta cells and remains stable thereafter. Progressive gain of beta-cell function occurs over 7 months, as measured by glucose-regulated insulin release and suppression of hyperglycemia. These studies demonstrate that lineage-reprogrammed cells persist for >1 year and undergo epigenetic, transcriptional, anatomical and functional development toward a beta-cell phenotype.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Insulin+ cells reprogrammed from pancreatic acinar cells can persist long-term in vivo.
Figure 2: Progressive formation of islet-like structures by induced insulin+ cells.
Figure 3: Induced beta cells acquire function over time.
Figure 4: DNA methylation dynamics of the induced beta cells.
Figure 5: Transcription network remodeling and stabilization within the induced beta cells.
Figure 6: Cellular and molecular milestones in the evolution of induced beta cells.

Accession codes

Primary accessions

Gene Expression Omnibus

Change history

  • 14 May 2015

    In the version of this article initially published, Yingying Zhang's name was spelled Yinying Zhang. The error has been corrected in the HTML and PDF versions of the article.

References

  1. 1

    Graf, T. & Enver, T. Forcing cells to change lineages. Nature 462, 587–594 (2009).

    CAS  Article  Google Scholar 

  2. 2

    Zhou, Q., Brown, J., Kanarek, A., Rajagopal, J. & Melton, D.A. In vivo reprogramming of adult pancreatic exocrine cells to beta-cells. Nature 455, 627–632 (2008).

    CAS  Article  Google Scholar 

  3. 3

    Vierbuchen, T. et al. Direct conversion of fibroblasts to functional neurons by defined factors. Nature 463, 1035–1041 (2010).

    CAS  Article  Google Scholar 

  4. 4

    Ieda, M. et al. Direct reprogramming of fibroblasts into functional cardiomyocytes by defined factors. Cell 142, 375–386 (2010).

    CAS  Article  Google Scholar 

  5. 5

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

    CAS  Article  Google Scholar 

  6. 6

    Ginsberg, M. et al. Efficient direct reprogramming of mature amniotic cells into endothelial cells by ETS factors and TGFβ suppression. Cell 151, 559–575 (2012).

    CAS  Article  Google Scholar 

  7. 7

    Al-Hasani, K. et al. Adult duct-lining cells can reprogram into beta-like cells able to counter repeated cycles of toxin-induced diabetes. Dev. Cell 26, 86–100 (2013).

    CAS  Article  Google Scholar 

  8. 8

    Guo, Z. et al. In vivo direct reprogramming of reactive glial cells into functional neurons after brain injury and in an Alzheimer's disease model. Cell Stem Cell 14, 188–202 (2014).

    CAS  Article  Google Scholar 

  9. 9

    Baeyens, L. et al. Transient cytokine treatment induces acinar cell reprogramming and regenerates functional beta cell mass in diabetic mice. Nat. Biotechnol. 32, 76–83 (2014).

    CAS  Article  Google Scholar 

  10. 10

    Lee, J. et al. Expansion and conversion of human pancreatic ductal cells into insulin-secreting endocrine cells. eLife 2, e00940 (2013).

    Article  Google Scholar 

  11. 11

    Vierbuchen, T. & Wernig, M. Molecular roadblocks for cellular reprogramming. Mol. Cell 47, 827–838 (2012).

    CAS  Article  Google Scholar 

  12. 12

    Chen, J.X. et al. Inefficient reprogramming of fibroblasts into cardiomyocytes using Gata4, Mef2c, and Tbx5. Circ. Res. 111, 50–55 (2012).

    CAS  Article  Google Scholar 

  13. 13

    Shaner, N.C., Steinbach, P.A. & Tsien, R.Y. A guide to choosing fluorescent proteins. Nat. Methods 2, 905–909 (2005).

    CAS  Article  Google Scholar 

  14. 14

    Teta, M., Long, S.Y., Wartschow, L.M., Rankin, M.M. & Kushner, J.A. Very slow turnover of beta-cells in aged adult mice. Diabetes 54, 2557–2567 (2005).

    CAS  Article  Google Scholar 

  15. 15

    Brennand, K., Huangfu, D. & Melton, D. All β cells contribute equally to islet growth and maintenance. PLoS Biol. 5, e163 (2007).

    Article  Google Scholar 

  16. 16

    Montaña, E., Bonner-Weir, S. & Weir, G.C. Transplanted beta cell response to increased metabolic demand. Changes in beta cell replication and mass. J. Clin. Invest. 93, 1577–1582 (1994).

    Article  Google Scholar 

  17. 17

    Bonner-Weir, S. Life and death of the pancreatic β cells. Trends Endocrinol. Metab. 11, 375–378 (2000).

    CAS  Article  Google Scholar 

  18. 18

    Juang, J.H., Bonner-Weir, S., Wu, Y.J. & Weir, G.C. Beneficial influence of glycemic control upon the growth and function of transplanted islets. Diabetes 43, 1334–1339 (1994).

    CAS  Article  Google Scholar 

  19. 19

    Montaña, E., Bonner-Weir, S. & Weir, G.C. Beta cell mass and growth after syngeneic islet cell transplantation in normal and streptozocin diabetic C57BL/6 mice. J. Clin. Invest. 91, 780–787 (1993).

    Article  Google Scholar 

  20. 20

    Meissner, A. et al. Genome-scale DNA methylation maps of pluripotent and differentiated cells. Nature 454, 766–770 (2008).

    CAS  Article  Google Scholar 

  21. 21

    Li, W. et al. In vivo reprogramming of pancreatic acinar cells to three islet endocrine subtypes. eLife 3, e01846 (2014).

    Article  Google Scholar 

  22. 22

    Blum, B. 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).

    CAS  Article  Google Scholar 

  23. 23

    Dahl, U., Sjodin, A. & Semb, H. Cadherins regulate aggregation of pancreatic beta-cells in vivo. Development 122, 2895–2902 (1996).

    CAS  PubMed  Google Scholar 

  24. 24

    Hopcroft, D.W., Mason, D.R. & Scott, R.S. Structure-function relationships in pancreatic islets: support for intraislet modulation of insulin secretion. Endocrinology 117, 2073–2080 (1985).

    CAS  Article  Google Scholar 

  25. 25

    Meda, P. et al. Rapid and reversible secretion changes during uncoupling of rat insulin-producing cells. J. Clin. Invest. 86, 759–768 (1990).

    CAS  Article  Google Scholar 

  26. 26

    Ravier, M.A. et al. Loss of connexin36 channels alters beta-cell coupling, islet synchronization of glucose-induced Ca2+ and insulin oscillations, and basal insulin release. Diabetes 54, 1798–1807 (2005).

    CAS  Article  Google Scholar 

  27. 27

    Konstantinova, I. et al. EphA-Ephrin-A-mediated beta cell communication regulates insulin secretion from pancreatic islets. Cell 129, 359–370 (2007).

    CAS  Article  Google Scholar 

  28. 28

    Akinci, E., Banga, A., Greder, L.V., Dutton, J.R. & Slack, J.M. Reprogramming of pancreatic exocrine cells towards a beta (β) cell character using Pdx1, Ngn3 and MafA. Biochem. J. 442, 539–550 (2012).

    CAS  Article  Google Scholar 

  29. 29

    Banga, A., Akinci, E., Greder, L.V., Dutton, J.R. & Slack, J.M. In vivo reprogramming of Sox9+ cells in the liver to insulin-secreting ducts. Proc. Natl. Acad. Sci. USA 109, 15336–15341 (2012).

    CAS  Article  Google Scholar 

  30. 30

    Chen, Y.J. et al. De novo formation of insulin-producing “neo-beta cell islets” from intestinal crypts. Cell Reports 6, 1046–1058 (2014).

    CAS  Article  Google Scholar 

  31. 31

    Gotoh, M. et al. Reproducible high yield of rat islets by stationary in vitro digestion following pancreatic ductal or portal venous collagenase injection. Transplantation 43, 725–730 (1987).

    CAS  Article  Google Scholar 

  32. 32

    Cavelti-Weder, C., Li, W., Weir, G.C. & Zhou, Q. Direct lineage conversion of pancreatic exocrine to endocrine beta cells in vivo with defined factors. Methods Mol. Biol. 1150, 247–262 (2014).

    CAS  Article  Google Scholar 

  33. 33

    Gu, H. et al. Preparation of reduced representation bisulfite sequencing libraries for genome-scale DNA methylation profiling. Nat. Protoc. 6, 468–481 (2011).

    CAS  Article  Google Scholar 

  34. 34

    Smyth, G.K. Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Genet. Molec. Biol. 3, 3 (2004).

    Article  Google Scholar 

Download references

Acknowledgements

We thank M. Shear, J. Brown, J. Hollister-Lock, and R. Zhao for technical assistance; H.-H. Chen for assistance with gene profiling; D. Yu and T. Fabbro for statistical analysis; Boston Children's Hospital core facility for an Illumina array; Joslin Specialized Assay Core for insulin measurement; members of the Zhou laboratory for advice and feedback; and D. Melton and D. Breault for discussion and reading of the manuscript. Q.Z. was supported by awards from the US National Institutes of Health. W.L. is supported by a postdoctoral fellowship from the Juvenile Diabetes Research Foundation. C.C.-W. is supported by postdoctoral fellowships from the Swiss Science Foundation and the Swiss Foundation for Grants in Biology and Medicine.

Author information

Affiliations

Authors

Contributions

Experimental design: W.L., C.C.-W., G.W., A.M., D.G., Q.Z. Experimental execution: W.L., C.C.-W., Yinying Z., S.D., G.G., M.S., K.X., T.H., T.Y., M.N., Yuemei Z., S.Z. Data analyses: W.L., C.C.-W., K.C., J.Z., M.S., T.H., A.M., G.W., Q.Z. Figure preparation, manuscript writing and editing: W.L., C.C.-W., K.C., G.W., A.M., Q.Z.

Corresponding author

Correspondence to Qiao Zhou.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–10 and Supplementary Tables 1 and 2 (PDF 1745 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Li, W., Cavelti-Weder, C., Zhang, Y. et al. Long-term persistence and development of induced pancreatic beta cells generated by lineage conversion of acinar cells. Nat Biotechnol 32, 1223–1230 (2014). https://doi.org/10.1038/nbt.3082

Download citation

Further reading

Search

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing