The Effect of Wnt Pathway Modulators on Human IPSC-Derived Pancreatic Beta Cell Maturation

Overview

Journal Front Endocrinol (Lausanne)

Specialty Endocrinology

Date 2019 May 30

PMID 31139151

Citations 23

Authors

Heidrun Vethe

Luiza Ghila

Magnus Berle

Laurence Hoareau

Oystein A Haaland

Hanne Scholz

Joao A Paulo

Simona Chera

Helge Raeder

Affiliations

Soon will be listed here.

Abstract

Current published protocols for targeted differentiation of human stem cells toward pancreatic β-cells fail to deliver sufficiently mature cells with functional properties comparable to human islet β-cells. We aimed to assess whether Wnt-modulation could promote the final protocol stages of β-cell maturation, building our hypothesis on our previous findings of Wnt activation in immature hiPSC-derived stage 7 (S7) cells compared to adult human islets and with recent data reporting a link between Wnt/PCP and β-cell maturation. In this study, we stimulated canonical and non-canonical Wnt signaling in hiPSC-derived S7 cells using syntetic proteins including WNT3A, WNT4, WNT5A and WNT5B, and we inhibited endogenous Wnt signaling with the Tankyrase inhibitor G007-LK (TKi). Whereas neither canonical nor non-canonical Wnt stimulation alone was able to mature hiPSC-derived S7 cells, WNT-inhibition with TKi increased the fraction of monohormonal cells and global proteomics of TKi-treated S7 cells showed a proteomic signature more similar to adult human islets, suggesting that inhibition of endogenous Wnt contributes toward final β-cell maturation.

Citing Articles

Tankyrase inhibition promotes endocrine commitment of hPSC-derived pancreatic progenitors.

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Cell identity dynamics and insight into insulin secretagogues when employing stem cell-derived islets for disease modeling.

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Heterozygous missense variant in GLI2 impairs human endocrine pancreas development.

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Targeted Gene Silencing by Using GapmeRs in Differentiating Human-Induced Pluripotent Stem Cells (hiPSC) Toward Pancreatic Progenitors.

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References

Topol L, Jiang X, Choi H, Garrett-Beal L, Carolan P, Yang Y . Wnt-5a inhibits the canonical Wnt pathway by promoting GSK-3-independent beta-catenin degradation. J Cell Biol. 2003; 162(5):899-908. PMC: 2172823. DOI: 10.1083/jcb.200303158. View

Veeman M, Axelrod J, Moon R . A second canon. Functions and mechanisms of beta-catenin-independent Wnt signaling. Dev Cell. 2003; 5(3):367-77. DOI: 10.1016/s1534-5807(03)00266-1. View

Nelson W, Nusse R . Convergence of Wnt, beta-catenin, and cadherin pathways. Science. 2004; 303(5663):1483-7. PMC: 3372896. DOI: 10.1126/science.1094291. View

Logan C, Nusse R . The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol. 2004; 20:781-810. DOI: 10.1146/annurev.cellbio.20.010403.113126. View

Mikels A, Nusse R . Purified Wnt5a protein activates or inhibits beta-catenin-TCF signaling depending on receptor context. PLoS Biol. 2006; 4(4):e115. PMC: 1420652. DOI: 10.1371/journal.pbio.0040115. View

Gordon M, Nusse R . Wnt signaling: multiple pathways, multiple receptors, and multiple transcription factors. J Biol Chem. 2006; 281(32):22429-33. DOI: 10.1074/jbc.R600015200. View

Beausoleil S, Villen J, Gerber S, Rush J, Gygi S . A probability-based approach for high-throughput protein phosphorylation analysis and site localization. Nat Biotechnol. 2006; 24(10):1285-92. DOI: 10.1038/nbt1240. View

Elias J, Gygi S . Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry. Nat Methods. 2007; 4(3):207-14. DOI: 10.1038/nmeth1019. View

Rulifson I, Karnik S, Heiser P, Ten Berge D, Chen H, Gu X . Wnt signaling regulates pancreatic beta cell proliferation. Proc Natl Acad Sci U S A. 2007; 104(15):6247-52. PMC: 1847455. DOI: 10.1073/pnas.0701509104. View

10.

Nusse R . Wnt signaling and stem cell control. Cell Res. 2008; 18(5):523-7. DOI: 10.1038/cr.2008.47. View

11.

Schinner S . Wnt-signalling and the metabolic syndrome. Horm Metab Res. 2009; 41(2):159-63. DOI: 10.1055/s-0028-1119408. View

12.

Komiya Y, Habas R . Wnt signal transduction pathways. Organogenesis. 2009; 4(2):68-75. PMC: 2634250. DOI: 10.4161/org.4.2.5851. View

13.

Friberg A, Stahle M, Brandhorst H, Korsgren O, Brandhorst D . Human islet separation utilizing a closed automated purification system. Cell Transplant. 2009; 17(12):1305-13. DOI: 10.3727/096368908787648100. View

14.

Wisniewski J, Zougman A, Nagaraj N, Mann M . Universal sample preparation method for proteome analysis. Nat Methods. 2009; 6(5):359-62. DOI: 10.1038/nmeth.1322. View

15.

Dorrell C, Abraham S, Lanxon-Cookson K, Canaday P, Streeter P, Grompe M . Isolation of major pancreatic cell types and long-term culture-initiating cells using novel human surface markers. Stem Cell Res. 2009; 1(3):183-94. DOI: 10.1016/j.scr.2008.04.001. View

16.

Elias J, Gygi S . Target-decoy search strategy for mass spectrometry-based proteomics. Methods Mol Biol. 2009; 604:55-71. PMC: 2922680. DOI: 10.1007/978-1-60761-444-9_5. View

17.

Huttlin E, Jedrychowski M, Elias J, Goswami T, Rad R, Beausoleil S . A tissue-specific atlas of mouse protein phosphorylation and expression. Cell. 2010; 143(7):1174-89. PMC: 3035969. DOI: 10.1016/j.cell.2010.12.001. View

18.

Nostro M, Sarangi F, Ogawa S, Holtzinger A, Corneo B, Li X . Stage-specific signaling through TGFβ family members and WNT regulates patterning and pancreatic specification of human pluripotent stem cells. Development. 2011; 138(5):861-71. PMC: 3035090. DOI: 10.1242/dev.055236. View

19.

Sokol S . Maintaining embryonic stem cell pluripotency with Wnt signaling. Development. 2011; 138(20):4341-50. PMC: 3177306. DOI: 10.1242/dev.066209. View

20.

Riedel M, Asadi A, Wang R, Ao Z, Warnock G, Kieffer T . Immunohistochemical characterisation of cells co-producing insulin and glucagon in the developing human pancreas. Diabetologia. 2011; 55(2):372-81. DOI: 10.1007/s00125-011-2344-9. View