Letter | Published:

Identification of proliferative and mature β-cells in the islets of Langerhans

Nature 535, 430–434 (21 July 2016) | Download Citation

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Abstract

Insulin-dependent diabetes is a complex multifactorial disorder characterized by loss or dysfunction of β-cells. Pancreatic β-cells differ in size, glucose responsiveness, insulin secretion and precursor cell potential1,2,3,4,5; understanding the mechanisms that underlie this functional heterogeneity might make it possible to develop new regenerative approaches. Here we show that Fltp (also known as Flattop and Cfap126), a Wnt/planar cell polarity (PCP) effector and reporter gene6, acts as a marker gene that subdivides endocrine cells into two subpopulations and distinguishes proliferation-competent from mature β-cells with distinct molecular, physiological and ultrastructural features. Genetic lineage tracing revealed that endocrine subpopulations from Fltp-negative and -positive lineages react differently to physiological and pathological changes. The expression of Fltp increases when endocrine cells cluster together to form polarized and mature 3D islet mini-organs7,8,9. We show that 3D architecture and Wnt/PCP ligands are sufficient to trigger β-cell maturation. By contrast, the Wnt/PCP effector Fltp is not necessary for β-cell development, proliferation or maturation. We conclude that 3D architecture and Wnt/PCP signalling underlie functional β-cell heterogeneity and induce β-cell maturation. The identification of Fltp as a marker for endocrine subpopulations sheds light on the molecular underpinnings of islet cell heterogeneity and plasticity and might enable targeting of endocrine subpopulations for the regeneration of functional β-cell mass in diabetic patients.

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Gene Expression Omnibus

Data deposits

Microarray data have been submitted to GEO with accession number GSE68853.

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Acknowledgements

We thank A. Raducanu, A. Böttcher and E. Schlüssel for comments and discussions; A. Theis, B. Vogel and K. Diemer for technical support; the Human Tissue Laboratory (HTL) of Lund University Diabetes Centre (LUDC) for high-quality RNA sequencing expression data from human pancreatic islets donors; to InSphero for human microislets; the European Consortium for Islet Transplantation (ECIT) for human islets; and R. Scharfmann for the EndoC-β H1 cell line. The human islet research was supported by a Strategic Research Grant from the Swedish Research Council (2009-1039). A.M. was funded by the Helmholtz post-doctoral fellowship program. This work was supported by an Emmy-Noether Fellowship, the European Union (ERC starting grant Ciliary Disease) and the HumEn project from the European Union's Seventh Framework Programme for Research, Technological Development and Demonstration under grant agreement No. 602587 (http://www.hum-en.eu/). This work was funded (in part) for H.L. and J.B. by the Helmholtz Alliance ICEMED – Imaging and Curing Environmental Metabolic Diseases, through the Initiative and Networking Fund of the Helmholtz Association. We thank the Helmholtz Society, Helmholtz Portfolio Theme 'Metabolic Dysfunction and Common Disease, German Research Foundation and German Center for Diabetes Research (DZD e.V.) for financial support. This work was supported with funds for S.S. from the Emmy-Noether Program, the Center for Regenerative Therapies Dresden-DFG Research Center for Regenerative Therapies Dresden, Cluster of Excellence (CRTD), the DFG-Collaborative Research Center/Transregio 127 and the German Ministry for Education and Research to the German Centre for Diabetes Research and to the Network of Competence for Diabetes.

Author information

    • Erik Bader
    •  & Adriana Migliorini

    These authors contributed equally to this work.

Affiliations

  1. Institute of Diabetes and Regeneration Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany

    • Erik Bader
    • , Adriana Migliorini
    • , Moritz Gegg
    • , Noah Moruzzi
    • , Jantje Gerdes
    • , Sara S. Roscioni
    • , Mostafa Bakhti
    • , Elisabeth Brandl
    •  & Heiko Lickert

  2. Institute of Stem Cell Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany

    • Erik Bader
    • , Adriana Migliorini
    • , Moritz Gegg
    •  & Heiko Lickert

  3. Institute of Epidemiology II, Helmholtz Zentrum München, 85764 Neuherberg, Germany

    • Erik Bader
    •  & Rui Wang-Sattler

  4. Department of Molecular Medicine and Surgery, Karolinska University Hospital, SE-17176 Stockholm, Sweden

    • Noah Moruzzi

  5. German Center for Diabetes Research (DZD), 85764 Neuherberg, Germany

    • Martin Irmler
    • , Johannes Beckers
    • , Martin Jastroch
    • , Matthias Tschöp
    • , Fausto Machicao
    • , Harald Staiger
    • , Hans-Ulrich Häring
    • , Helena Chmelova
    • , Julie A. Chouinard
    • , Stephan Speier
    •  & Heiko Lickert

  6. Institute of Experimental Genetics, Helmholtz Zentrum München, 85764 Neuherberg, Germany

    • Martin Irmler
    •  & Johannes Beckers

  7. Technische Universität München, Ismaninger Straße 22, 81675 München, Germany

    • Johannes Beckers
    •  & Heiko Lickert

  8. Research Unit Analytical Pathology, Helmholtz Zentrum München, 85764 Neuherberg, Germany

    • Michaela Aichler
    •  & Annette Feuchtinger

  9. Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München, 85764 Neuherberg, Germany

    • Christin Leitzinger
    •  & Hans Zischka

  10. Institute of Diabetes and Obesity, Helmholtz Zentrum München, 85764 Neuherberg, Germany

    • Martin Jastroch
    •  & Matthias Tschöp

  11. Institute for Diabetes Research and Metabolic Diseases, Helmholtz Zentrum München, University of Tübingen, 72076 Tübingen, Germany

    • Fausto Machicao
    • , Harald Staiger
    •  & Hans-Ulrich Häring

  12. Department of Internal Medicine, Division of Endocrinology, Diabetology, Vascular Disease, Nephrology and Clinical Chemistry, University of Tübingen, 72076 Tübingen, Germany

    • Harald Staiger
    •  & Hans-Ulrich Häring

  13. Paul Langerhans Institute Dresden (PLID), Helmholtz Zentrum München, University Clinic Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany

    • Helena Chmelova
    • , Julie A. Chouinard
    •  & Stephan Speier

  14. DFG-Center for Regenerative Therapies Dresden (CRTD), Faculty of Medicine, Technische Universität Dresden, 01307 Dresden, Germany

    • Helena Chmelova
    • , Julie A. Chouinard
    •  & Stephan Speier

  15. Diabetes and Endocrinology, Lund University Diabetes Centre, 205 02 Malmö, Sweden

    • Nikolay Oskolkov

  16. Department of Immunology, Genetics and Pathology, Uppsala University, 751 05 Uppsala, Sweden

    • Olle Korsgren

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Contributions

E.B., A.M. and M.G. conceived and performed the experiments and wrote the manuscript. M.B. established the 3D in vitro cultures. S.S.R. and E.B. helped to perform human islet experiments. N.M., C.L., H.Z., M.J., M.T. and J.G. conceived and performed mitochondria analysis. M.I. and J.B. performed the microarray analysis. M.A. and A.F. analysed mitochondria by TEM. F.M., H.S. and H-U.H. performed the SNP analysis of FLTP in humans. O.K. provided human islets and N.O. provided RNA sequencing data. R.W-S. supervised and trained E.B. in human epidemiology and genetics. H.C., J.A.C. and S.S. performed the islet transplantation and analyzed the data set. H.L. conceived the study and wrote the manuscript.

Competing interests

H.L., M.G., E.B. and A.M. are inventors of a European Patent (PCT/EP2015/056664) based on this work.

Corresponding author

Correspondence to Heiko Lickert.

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

Extended data

Extended data figures

  1. 1.

    FVR expression is induced in compacted islet structures and correlates with high Nkx6.1 expression.

  2. 2.

    FVR− and FVR+ β-cell subpopulations exhibit differential proliferative capacity and inter-islet heterogeneity in the head and tail of the pancreas.

  3. 3.

    Validation of the sorting scheme and molecular and cellular analysis of FVR− and FVR+ endocrine subpopulations.

  4. 4.

    Transmission electron microscopy (TEM) analysis of Fltp− and Fltp+ β-cell subpopulations and quantification of mitochondria membrane length.

  5. 5.

    Analysis of oxygen consumption rate (OCR), mitochondria network and GSIS in FVR+ and FVR− β-cells.

  6. 6.

    Fltp genetic lineage tracing of endocrine subpopulation dynamics upon islet transplantation and HFD.

  7. 7.

    3D architecture and Wnt/PCP induce β-cell maturation in Min6 cells.

  8. 8.

    Modulation of maturation and proliferation through WNT signalling in human microislets and EndoC-β H1 human β-cell line and SNP association with insulin secretion defects in the FLTP gene.

  9. 9.

    Knockout of Fltp has no effect on β-cell maturation or proliferation.

  10. 10.

    Analysis of the metabolic phenotype of FltpZV/ZV mice.

Supplementary information

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  1. 1.

    Supplementary Information

    This file contains the Raw Data files for Figures 2k, 4h, 4j and Extended Data Figures 7m and 8f.

  2. 2.

    Supplementary Tables

    This file contains Supplementary Tables 1-5.

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DOI

https://doi.org/10.1038/nature18624

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