Wolfram syndrome 1 gene regulates pathways maintaining beta-cell health and survival

Abstract

Wolfram Syndrome 1 (WFS1) protein is an endoplasmic reticulum (ER) factor whose deficiency results in juvenile-onset diabetes secondary to cellular dysfunction and apoptosis. The mechanisms guiding β-cell outcomes secondary to WFS1 function, however, remain unclear. Here, we show that WFS1 preserves normal β-cell physiology by promoting insulin biosynthesis and negatively regulating ER stress. Depletion of Wfs1 in vivo and in vitro causes functional defects in glucose-stimulated insulin secretion and insulin content, triggering Chop-mediated apoptotic pathways. Genetic proof of concept studies coupled with RNA-seq reveal that increasing WFS1 confers a functional and a survival advantage to β-cells under ER stress by increasing insulin gene expression and downregulating the Chop-Trib3 axis, thereby activating Akt pathways. Remarkably, WFS1 and INS levels are reduced in type-2 diabetic (T2DM) islets, suggesting that WFS1 may contribute to T2DM β-cell pathology. Taken together, this work reveals essential pathways regulated by WFS1 to control β-cell survival and function primarily through preservation of ER homeostasis.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1: Glucose tolerance and β-cell morphometry in whole body WFS1–/– 129S6 mice.
Fig. 2: WFS1 positively regulates insulin biosynthesis and β-cell maturity markers.
Fig. 3: WFS1 overexpression suppresses ER stress-mediated cell death.
Fig. 4: WFS1 knockdown promotes β-cell dysfunction, ER stress, and β-cell death.
Fig. 5: WFS1 expression is decreased in type-2 diabetic human donor islets.

References

  1. 1.

    Cnop M, Welsh N, Jonas JC, Jorns A, Lenzen S, Eizirik DL. Mechanisms of pancreatic beta-cell death in type 1 and type 2 diabetes: many differences, few similarities. Diabetes. 2005;54:S97–107.

    CAS  PubMed  Article  Google Scholar 

  2. 2.

    Donath MY, Ehses JA, Maedler K, Schumann DM, Ellingsgaard H, Eppler E, et al. Mechanisms of beta-cell death in type 2 diabetes. Diabetes. 2005;54:S108–13.

    CAS  PubMed  Article  Google Scholar 

  3. 3.

    Back SH, Kaufman RJ. Endoplasmic reticulum stress and type 2 diabetes. Annu Rev Biochem. 2012;81:767–93.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  4. 4.

    Harding HP, Ron D. Endoplasmic reticulum stress and the development of diabetes: a review. Diabetes. 2002;51:S455–61.

    CAS  PubMed  Article  Google Scholar 

  5. 5.

    Inoue H, Tanizawa Y, Wasson J, Behn P, Kalidas K, Bernal-Mizrachi E, et al. A gene encoding a transmembrane protein is mutated in patients with diabetes mellitus and optic atrophy (Wolfram syndrome). Nat Genet. 1998;20:143–8.

    CAS  PubMed  Article  Google Scholar 

  6. 6.

    Urano F. Wolfram syndrome: diagnosis, management, and treatment. Curr Diabetes Rep. 2016;16:6.

    Article  Google Scholar 

  7. 7.

    Sandhu MS, Weedon MN, Fawcett KA, Wasson J, Debenham SL, Daly A, et al. Common variants in WFS1 confer risk of type 2 diabetes. Nat Genet. 2007;39:951–3.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  8. 8.

    Bonnycastle LL, Chines PS, Hara T, Huyghe JR, Swift AJ, Heikinheimo P, et al. Autosomal dominant diabetes arising from a Wolfram syndrome 1 mutation. Diabetes. 2013;62:3943–50.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  9. 9.

    Fonseca SG, Fukuma M, Lipson KL, Nguyen LX, Allen JR, Oka Y, et al. WFS1 is a novel component of the unfolded protein response and maintains homeostasis of the endoplasmic reticulum in pancreatic beta-cells. J Biol Chem. 2005;280:39609–15.

    CAS  PubMed  Article  Google Scholar 

  10. 10.

    Fonseca SG, Ishigaki S, Oslowski CM, Lu S, Lipson KL, Ghosh R, et al. Wolfram syndrome 1 gene negatively regulates ER stress signaling in rodent and human cells. J Clin Investig. 2010;120:744–55.

    CAS  PubMed  Article  Google Scholar 

  11. 11.

    Lu S, Kanekura K, Hara T, Mahadevan J, Spears LD, Oslowski CM, et al. A calcium-dependent protease as a potential therapeutic target for Wolfram syndrome. Proc Natl Acad Sci USA. 2014;111:E5292–301.

    CAS  PubMed  Article  Google Scholar 

  12. 12.

    Luuk H, Koks S, Plaas M, Hannibal J, Rehfeld JF, Vasar E. Distribution of Wfs1 protein in the central nervous system of the mouse and its relation to clinical symptoms of the Wolfram syndrome. J Comp Neurol. 2008;509:642–60.

    CAS  PubMed  Article  Google Scholar 

  13. 13.

    Wang Z, York NW, Nichols CG, Remedi MS. Pancreatic beta cell dedifferentiation in diabetes and redifferentiation following insulin therapy. Cell Metab. 2014;19:872–82.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  14. 14.

    Hohmeier HE, Mulder H, Chen G, Henkel-Rieger R, Prentki M, Newgard CB. Isolation of INS-1-derived cell lines with robust ATP-sensitive K+ channel-dependent and -independent glucose-stimulated insulin secretion. Diabetes. 2000;49:424–30.

    CAS  PubMed  Article  Google Scholar 

  15. 15.

    Clark AL, Kanekura K, Lavagnino Z, Spears LD, Abreu D, Mahadevan J, et al. Targeting cellular calcium homeostasis to prevent cytokine-mediated beta cell death. Sci Rep. 2017;7:5611.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  16. 16.

    Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2013;29:15–21.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  17. 17.

    Liao Y, Smyth GK, Shi W. featureCounts: an efficient general purpose program for assigning sequence reads to genomic features. Bioinformatics. 2014;30:923–30.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  18. 18.

    Patro R, Duggal G, Love MI, Irizarry RA, Kingsford C. Salmon provides fast and bias-aware quantification of transcript expression. Nat Methods. 2017;14:417–9.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  19. 19.

    Wang L, Wang S, Li W. RSeQC: quality control of RNA-seq experiments. Bioinformatics. 2012;28:2184–5.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  20. 20.

    Robinson MD, McCarthy DJ, Smyth GK. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 2010;26:139–40.

    CAS  Article  Google Scholar 

  21. 21.

    Ritchie ME, Phipson B, Wu D, Hu Y, Law CW, Shi W, et al. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 2015;43:e47.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  22. 22.

    Liu R, Holik AZ, Su S, Jansz N, Chen K, Leong HS, et al. Why weight? Modelling sample and observational level variability improves power in RNA-seq analyses. Nucleic Acids Res. 2015;43:e97.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  23. 23.

    Luo W, Friedman MS, Shedden K, Hankenson KD, Woolf PJ. GAGE: generally applicable gene set enrichment for pathway analysis. BMC Bioinform. 2009;10:161.

    Article  CAS  Google Scholar 

  24. 24.

    Ishihara H, Takeda S, Tamura A, Takahashi R, Yamaguchi S, Takei D, et al. Disruption of the WFS1 gene in mice causes progressive beta-cell loss and impaired stimulus-secretion coupling in insulin secretion. Hum Mol Genet. 2004;13:1159–70.

    CAS  PubMed  Article  Google Scholar 

  25. 25.

    Riggs AC, Bernal-Mizrachi E, Ohsugi M, Wasson J, Fatrai S, Welling C, et al. Mice conditionally lacking the Wolfram gene in pancreatic islet beta cells exhibit diabetes as a result of enhanced endoplasmic reticulum stress and apoptosis. Diabetologia. 2005;48:2313–21.

    CAS  PubMed  Article  Google Scholar 

  26. 26.

    Yamada T, Ishihara H, Tamura A, Takahashi R, Yamaguchi S, Takei D, et al. WFS1-deficiency increases endoplasmic reticulum stress, impairs cell cycle progression and triggers the apoptotic pathway specifically in pancreatic beta-cells. Hum Mol Genet. 2006;15:1600–9.

    CAS  PubMed  Article  Google Scholar 

  27. 27.

    Du K, Herzig S, Kulkarni RN, Montminy M. TRB3: a tribbles homolog that inhibits Akt/PKB activation by insulin in liver. Science. 2003;300:1574–7.

    CAS  PubMed  Article  Google Scholar 

  28. 28.

    Takei D, Ishihara H, Yamaguchi S, Yamada T, Tamura A, Katagiri H, et al. WFS1 protein modulates the free Ca(2+) concentration in the endoplasmic reticulum. FEBS Lett. 2006;580:5635–40.

    CAS  PubMed  Article  Google Scholar 

  29. 29.

    Hara T, Mahadevan J, Kanekura K, Hara M, Lu S, Urano F. Calcium efflux from the endoplasmic reticulum leads to beta-cell death. Endocrinology. 2014;155:758–68.

    CAS  PubMed  Article  Google Scholar 

  30. 30.

    Butler AE, Janson J, Bonner-Weir S, Ritzel R, Rizza RA, Butler PC. Beta-cell deficit and increased beta-cell apoptosis in humans with type 2 diabetes. Diabetes. 2003;52:102–10.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  31. 31.

    Evans-Molina C, Hatanaka M, Mirmira RG. Lost in translation: endoplasmic reticulum stress and the decline of beta-cell health in diabetes mellitus. Diabetes Obes Metab. 2013;15:159–69.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  32. 32.

    Henquin JC, Ibrahim MM, Rahier J. Insulin, glucagon and somatostatin stores in the pancreas of subjects with type-2 diabetes and their lean and obese non-diabetic controls. Sci Rep. 2017;7:11015.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  33. 33.

    Noormets K, Koks S, Muldmaa M, Mauring L, Vasar E, Tillmann V. Sex differences in the development of diabetes in mice with deleted wolframin (Wfs1) gene. Exp Clin Endocrinol Diabetes. 2011;119:271–5.

    CAS  PubMed  Article  Google Scholar 

  34. 34.

    Ivask M, Hugill A, Koks S. RNA-sequencing of WFS1-deficient pancreatic islets. Physiol Rep. 2016;4:e12750.

  35. 35.

    de Heredia ML, Cleries R, Nunes V. Genotypic classification of patients with Wolfram syndrome: insights into the natural history of the disease and correlation with phenotype. Genet Med. 2013;15:497–506.

    PubMed  Article  CAS  Google Scholar 

  36. 36.

    Chaussenot A, Bannwarth S, Rouzier C, Vialettes B, Mkadem SA, Chabrol B, et al. Neurologic features and genotype-phenotype correlation in Wolfram syndrome. Ann Neurol. 2011;69:501–8.

    CAS  PubMed  Article  Google Scholar 

  37. 37.

    Cryns K, Sivakumaran TA, Van den Ouweland JM, Pennings RJ, Cremers CW, Flothmann K, et al. Mutational spectrum of the WFS1 gene in Wolfram syndrome, nonsyndromic hearing impairment, diabetes mellitus, and psychiatric disease. Hum Mutat. 2003;22:275–87.

    CAS  PubMed  Article  Google Scholar 

  38. 38.

    Scheuner D, Kaufman RJ. The unfolded protein response: a pathway that links insulin demand with beta-cell failure and diabetes. Endocr Rev. 2008;29:317–33.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  39. 39.

    Fonseca SG, Gromada J, Urano F. Endoplasmic reticulum stress and pancreatic beta-cell death. Trends Endocrinol Metab. 2011;22:266–74.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. 40.

    Lipson KL, Fonseca SG, Ishigaki S, Nguyen LX, Foss E, Bortell R, et al. Regulation of insulin biosynthesis in pancreatic beta cells by an endoplasmic reticulum-resident protein kinase IRE1. Cell Metab. 2006;4:245–54.

    CAS  PubMed  Article  Google Scholar 

  41. 41.

    Seo HY, Kim YD, Lee KM, Min AK, Kim MK, Kim HS, et al. Endoplasmic reticulum stress-induced activation of activating transcription factor 6 decreases insulin gene expression via up-regulation of orphan nuclear receptor small heterodimer partner. Endocrinology. 2008;149:3832–41.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  42. 42.

    Cissell MA, Zhao L, Sussel L, Henderson E, Stein R. Transcription factor occupancy of the insulin gene in vivo. Evidence for direct regulation by Nkx2.2. J Biol Chem. 2003;278:751–6.

    CAS  PubMed  Article  Google Scholar 

  43. 43.

    Gutierrez GD, Bender AS, Cirulli V, Mastracci TL, Kelly SM, Tsirigos A, et al. Pancreatic beta cell identity requires continual repression of non-beta cell programs. J Clin Investig. 2017;127:244–59.

    PubMed  Article  Google Scholar 

  44. 44.

    Le Lay J, Stein R. Involvement of PDX-1 in activation of human insulin gene transcription. J Endocrinol. 2006;188:287–94.

    PubMed  Article  CAS  Google Scholar 

  45. 45.

    Schaffer AE, Taylor BL, Benthuysen JR, Liu J, Thorel F, Yuan W, et al. Nkx6.1 controls a gene regulatory network required for establishing and maintaining pancreatic Beta cell identity. PLoS Genet. 2013;9:e1003274.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  46. 46.

    Shang L, Hua H, Foo K, Martinez H, Watanabe K, Zimmer M, et al. beta-cell dysfunction due to increased ER stress in a stem cell model of Wolfram syndrome. Diabetes. 2014;63:923–33.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  47. 47.

    Song B, Scheuner D, Ron D, Pennathur S, Kaufman RJ. Chop deletion reduces oxidative stress, improves beta cell function, and promotes cell survival in multiple mouse models of diabetes. J Clin Investig. 2008;118:3378–89.

    CAS  PubMed  Article  Google Scholar 

  48. 48.

    Szabat M, Page MM, Panzhinskiy E, Skovso S, Mojibian M, Fernandez-Tajes J, et al. Reduced insulin production relieves endoplasmic reticulum stress and induces beta cell proliferation. Cell Metab. 2016;23:179–93.

    CAS  PubMed  Article  Google Scholar 

  49. 49.

    Sharma RB, O’Donnell AC, Stamateris RE, Ha B, McCloskey KM, Reynolds PR, et al. Insulin demand regulates beta cell number via the unfolded protein response. J Clin Investig. 2015;125:3831–46.

    PubMed  Article  Google Scholar 

  50. 50.

    Porat S, Weinberg-Corem N, Tornovsky-Babaey S, Schyr-Ben-Haroush R, Hija A, Stolovich-Rain M, et al. Control of pancreatic beta cell regeneration by glucose metabolism. Cell Metab. 2011;13:440–9.

    CAS  PubMed  Article  Google Scholar 

  51. 51.

    Marchetti P, Bugliani M, Lupi R, Marselli L, Masini M, Boggi U, et al. The endoplasmic reticulum in pancreatic beta cells of type 2 diabetes patients. Diabetologia. 2007;50:2486–94.

    CAS  PubMed  Article  Google Scholar 

  52. 52.

    Laybutt DR, Preston AM, Akerfeldt MC, Kench JG, Busch AK, Biankin AV, et al. Endoplasmic reticulum stress contributes to beta cell apoptosis in type 2 diabetes. Diabetologia. 2007;50:752–63.

    CAS  PubMed  Article  Google Scholar 

  53. 53.

    Eizirik DL, Cnop M. ER stress in pancreatic beta cells: the thin red line between adaptation and failure. Sci Signal. 2010;3:pe7.

    PubMed  Article  CAS  Google Scholar 

  54. 54.

    Urano F. Wolfram syndrome iPS cells: the first human cell model of endoplasmic reticulum disease. Diabetes. 2014;63:844–6.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  55. 55.

    Papa FR. Endoplasmic reticulum stress, pancreatic beta-cell degeneration, and diabetes. Cold Spring Harb Perspect Med. 2012;2:a007666.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  56. 56.

    Stitzel ML, Sethupathy P, Pearson DS, Chines PS, Song L, Erdos MR, et al. Global epigenomic analysis of primary human pancreatic islets provides insights into type 2 diabetes susceptibility loci. Cell Metab. 2010;12:443–55.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

Download references

Acknowledgements

We thank the Genome Technology Access Center in the Department of Genetics at Washington University School of Medicine for help with genomic analysis. The Center is partially supported by NCI Cancer Center Support Grant P30CA91842 to the Siteman Cancer Center and by ICTS/CTSA Grant UL1TR000448 from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH), and NIH Roadmap for Medical Research. This publication is solely the responsibility of the authors and does not necessarily represent the official view of NCRR or NIH. The authors would like to acknowledge Cris Brown, Akari Takesato, and Lucas Peng for their technical assistance. This work was partly supported by the grants from the National Institutes of Health/NIDDK (DK112921, DK020579) and National Institutes of Health/NCATS (TR002065, TR000448) and philanthropic supports from the Unravel Wolfram Syndrome Fund, the Feiock Fund, the Silberman Fund, the Stowe Fund, the Ellie White Foundation for Rare Genetic Disorders, the Eye Hope Foundation, and the Snow Foundation to FU, DA was supported by the NIH training grant (F30DK111070).

Author information

Affiliations

Authors

Corresponding author

Correspondence to Fumihiko Urano.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Abreu, D., Asada, R., Revilla, J.M.P. et al. Wolfram syndrome 1 gene regulates pathways maintaining beta-cell health and survival. Lab Invest 100, 849–862 (2020). https://doi.org/10.1038/s41374-020-0408-5

Download citation

Further reading

Search

Quick links