, Volume 61, Issue 4, pp 906–918 | Cite as

SIRT6-mediated transcriptional suppression of Txnip is critical for pancreatic beta cell function and survival in mice

  • Kunhua Qin
  • Ning Zhang
  • Zhao Zhang
  • Michael Nipper
  • Zhenxin Zhu
  • Jake Leighton
  • Kexin Xu
  • Nicolas Musi
  • Pei Wang



Better understanding of how genetic and epigenetic components control beta cell differentiation and function is key to the discovery of novel therapeutic approaches to prevent beta cell dysfunction and failure in the progression of type 2 diabetes. Our goal was to elucidate the role of histone deacetylase sirtuin 6 (SIRT6) in beta cell development and homeostasis.


Sirt6 endocrine progenitor cell conditional knockout and beta cell-specific knockout mice were generated using the Cre-loxP system. Mice were assayed for islet morphology, glucose tolerance, glucose-stimulated insulin secretion and susceptibility to streptozotocin. Transcriptional regulatory functions of SIRT6 in primary islets were evaluated by RNA-Seq analysis. Reverse transcription-quantitative (RT-q)PCR and immunoblot were used to verify and investigate the gene expression changes. Chromatin occupancies of SIRT6, H3K9Ac, H3K56Ac and active RNA polymerase II were evaluated by chromatin immunoprecipitation.


Deletion of Sirt6 in pancreatic endocrine progenitor cells did not affect endocrine morphology, beta cell mass or insulin production but did result in glucose intolerance and defective glucose-stimulated insulin secretion in mice. Conditional deletion of Sirt6 in adult beta cells reproduced the insulin secretion defect. Loss of Sirt6 resulted in aberrant upregulation of thioredoxin-interacting protein (TXNIP) in beta cells. SIRT6 deficiency led to increased acetylation of histone H3 lysine residue at 9 (H3K9Ac), acetylation of histone H3 lysine residue at 56 (H3K56Ac) and active RNA polymerase II at the promoter region of Txnip. SIRT6-deficient beta cells exhibited a time-dependent increase in H3K9Ac, H3K56Ac and TXNIP levels. Finally, beta cell-specific SIRT6-deficient mice showed increased sensitivity to streptozotocin.


Our results reveal that SIRT6 suppresses Txnip expression in beta cells via deacetylation of histone H3 and plays a critical role in maintaining beta cell function and viability.

Data availability

Sequence data have been deposited in the National Institutes of Health (NIH) Gene Expression Omnibus (GEO) with the accession code GSE104161.


Beta cell Diabetes H3K9Ac Insulin secretion SIRT6 TXNIP 



Beta cell-specific knockout


Chromatin immunoprecipitation


Endocrine pancreas-specific knockout


Histone acetyltransferase


Histone deacetylase


Acetylation of histone H3 lysine residue at 9


Acetylation of histone H3 lysine residue at 18


Acetylation of histone H3 lysine residue at 56


Mouse insulin promoter 1-driven, inducible CreERT transgenic line


Neurogenin 3


National Institutes of Health


Reactive oxygen species


Sirtuin 6




Transcriptional start site


Thioredoxin-interacting protein



We thank Y. Liu (Department of Cell Systems & Anatomy, Greehey Children’s Cancer Research Institute, University of Texas Health Science Centre at San Antonio, USA) for assistance in analysing the RNA-Seq data and thank C. Cervantes (Department of Pharmacology, University of Texas Health Science Centre at San Antonio, USA) and C. Dong (Biochemistry Molecular Biology, Indiana University, USA) for critical reading of the manuscript.

Contribution statement

KQ and PW were responsible for designing the experiments. KQ, NZ, ZZ, MN, ZXZ and JL were responsible for acquisition of data. KQ, NZ, KX, NM and PW analysed and interpreted data. KQ drafted the manuscript. All authors critically revised the manuscript and approved the final version. PW is the guarantor of this work.


PW and KX are CPRIT Scholars and are supported by the Cancer Prevention and Research Institute of Texas. This work was supported by grants from the NIH (R01-DK80157 and R01-DK089229) and the American Diabetes Association to NM. This research also was supported by the San Antonio Nathan Shock Centre of Excellence in Aging Biology (P30 AG013319) and San Antonio Claude D. Pepper Older Americans Independence Centre (P30 AG044271).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2017_4542_MOESM1_ESM.pdf (769 kb)
ESM (PDF 769 kb)


  1. 1.
    Ashcroft FM, Rorsman P (2012) Diabetes mellitus and the beta cell: the last ten years. Cell 148:1160–1171CrossRefPubMedGoogle Scholar
  2. 2.
    Zaccardi F, Webb DR, Yates T, Davies MJ (2016) Pathophysiology of type 1 and type 2 diabetes mellitus: a 90-year perspective. Postgrad Med J 92:63–69CrossRefPubMedGoogle Scholar
  3. 3.
    Xie R, Carrano AC, Sander M (2015) A systems view of epigenetic networks regulating pancreas development and beta-cell function. Wiley Interdiscip Rev Syst Biol Med 7:1–11CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Bernstein D, Golson ML, Kaestner KH (2017) Epigenetic control of β-cell function and failure. Diabetes Res Clin Pract 123:24–36CrossRefPubMedGoogle Scholar
  5. 5.
    Arguelles AO, Meruvu S, Bowman JD, Choudhury M (2016) Are epigenetic drugs for diabetes and obesity at our door step? Drug Discov Today 21:499–509CrossRefPubMedGoogle Scholar
  6. 6.
    Haberland M, Montgomery RL, Olson EN (2009) The many roles of histone deacetylases in development and physiology: implications for disease and therapy. Nat Rev Genet 10:32–42CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Lee KK, Workman JL (2007) Histone acetyltransferase complexes: one size doesn’t fit all. Nat Rev Mol Cell Biol 8:284–295CrossRefPubMedGoogle Scholar
  8. 8.
    Remsberg JR, Ediger BN, Ho WY et al (2017) Deletion of histone deacetylase 3 in adult beta cells improves glucose tolerance via increased insulin secretion. Mol Metab 6:30–37CrossRefPubMedGoogle Scholar
  9. 9.
    Chang HC, Guarente L (2014) SIRT1 and other sirtuins in metabolism. Trends Endocrinol Metab 25:138–145CrossRefPubMedGoogle Scholar
  10. 10.
    Imai S, Guarente L (2014) NAD+ and sirtuins in aging and disease. Trends Cell Biol 24:464–471CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Michishita E, McCord RA, Berber E et al (2008) SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. Nature 452:492–496CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Yang B, Zwaans BM, Eckersdorff M, Lombard DB (2009) The sirtuin SIRT6 deacetylates H3 K56Ac in vivo to promote genomic stability. Cell Cycle 8:2662–2663CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Tasselli L, Xi Y, Zheng W et al (2016) SIRT6 deacetylates H3K18ac at pericentric chromatin to prevent mitotic errors and cellular senescence. Nat Struct Mol Biol 23:434–440CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Tasselli L, Zheng W, Chua KF (2017) SIRT6: novel mechanisms and links to aging and disease. Trends Endocrinol Metab 28:168–185CrossRefPubMedGoogle Scholar
  15. 15.
    Kim HS, Xiao C, Wang RH et al (2010) Hepatic-specific disruption of SIRT6 in mice results in fatty liver formation due to enhanced glycolysis and triglyceride synthesis. Cell Metab 12:224–236CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Schonhoff SE, Giel-Moloney M, Leiter AB (2004) Neurogenin 3-expressing progenitor cells in the gastrointestinal tract differentiate into both endocrine and non-endocrine cell types. Dev Biol 270:443–454CrossRefPubMedGoogle Scholar
  17. 17.
    Wicksteed B, Brissova M, Yan W et al (2010) Conditional gene targeting in mouse pancreatic ss-cells: analysis of ectopic Cre transgene expression in the brain. Diabetes 59:3090–3098CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Pan H, Qin K, Guo Z et al (2014) Negative elongation factor controls energy homeostasis in cardiomyocytes. Cell Rep 7:79–85CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Oropeza D, Jouvet N, Budry L et al (2015) Phenotypic characterization of MIP-CreERT1Lphi mice with transgene-driven islet expression of human growth hormone. Diabetes 64:3798–3807CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Zhang J, Zhang N, Liu M et al (2012) Disruption of growth factor receptor-binding protein 10 in the pancreas enhances beta-cell proliferation and protects mice from streptozotocin-induced beta-cell apoptosis. Diabetes 61:3189–3198CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Neuman JC, Truchan NA, Joseph JW, Kimple ME (2014) A method for mouse pancreatic islet isolation and intracellular cAMP determination. J Vis Exp e50374Google Scholar
  22. 22.
    Benitez CM, Qu K, Sugiyama T et al (2014) An integrated cell purification and genomics strategy reveals multiple regulators of pancreas development. PLoS Genet 10:e1004645CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Dahl JA, Collas P (2008) A rapid micro chromatin immunoprecipitation assay (microChIP). Nat Protoc 3:1032–1045CrossRefPubMedGoogle Scholar
  24. 24.
    Goodyer WR, Gu X, Liu Y, Bottino R, Crabtree GR, Kim SK (2012) Neonatal β cell development in mice and humans is regulated by calcineurin/NFAT. Dev Cell 23:21–34CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Xiong X, Wang G, Tao R et al (2016) Sirtuin 6 regulates glucose-stimulated insulin secretion in mouse pancreatic beta cells. Diabetologia 59:151–160CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Song MY, Wang J, Ka SO, Bae EJ, Park BH (2016) Insulin secretion impairment in Sirt6 knockout pancreatic β cells is mediated by suppression of the FoxO1-Pdx1-Glut2 pathway. Sci Rep 6:30321CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Ayala JE, Samuel VT, Morton GJ et al (2010) Standard operating procedures for describing and performing metabolic tests of glucose homeostasis in mice. Dis Model Mech 3:525–534CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Shalev A (2014) Minireview: thioredoxin-interacting protein: regulation and function in the pancreatic β-cell. Mol Endocrinol 28:1211–1220CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Yoshihara E, Fujimoto S, Inagaki N et al (2010) Disruption of TBP-2 ameliorates insulin sensitivity and secretion without affecting obesity. Nat Commun 1:127CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Rani S, Mehta JP, Barron N et al (2010) Decreasing Txnip mRNA and protein levels in pancreatic MIN6 cells reduces reactive oxygen species and restores glucose regulated insulin secretion. Cell Physiol Biochem 25:667–674CrossRefPubMedGoogle Scholar
  31. 31.
    Luo Y, He F, Hu L et al (2014) Transcription factor Ets1 regulates expression of thioredoxin-interacting protein and inhibits insulin secretion in pancreatic beta-cells. PLoS One 9:e99049CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Xu G, Chen J, Jing G, Shalev A (2012) Preventing β-cell loss and diabetes with calcium channel blockers. Diabetes 61:848–856CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    De Marinis Y, Cai M, Bompada P et al (2016) Epigenetic regulation of the thioredoxin-interacting protein (TXNIP) gene by hyperglycemia in kidney. Kidney Int 89:342–353CrossRefPubMedGoogle Scholar
  34. 34.
    Yu FX, Luo Y (2009) Tandem ChoRE and CCAAT motifs and associated factors regulate Txnip expression in response to glucose or adenosine-containing molecules. PLoS One 4:e8397CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Etchegaray JP, Chavez L, Huang Y et al (2015) The histone deacetylase SIRT6 controls embryonic stem cell fate via TET-mediated production of 5-hydroxymethylcytosine. Nat Cell Biol 17:545–557CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Reinert RB, Kantz J, Misfeldt AA et al (2012) Tamoxifen-induced Cre-loxP recombination is prolonged in pancreatic islets of adult mice. PLoS One 7:e33529CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Deeds MC, Anderson JM, Armstrong AS et al (2011) Single dose streptozotocin-induced diabetes: considerations for study design in islet transplantation models. Lab Anim 45:131–140CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Devis G, Somers G, Van Obberghen E, Malaisse WJ (1975) Calcium antagonists and islet function. I. Inhibition of insulin release by verapamil. Diabetes 24:247–251CrossRefPubMedGoogle Scholar
  39. 39.
    Semple CG, Smith M, Furman BL (1988) Inhibition of glucose-induced insulin secretion by calcium channel blocking drugs in-vitro but not in-vivo in the rat. J Pharm Pharmacol 40:22–26CrossRefPubMedGoogle Scholar
  40. 40.
    Semple CG, Thomson JA, Beastall GH, Lorimer AR (1983) Oral verapamil does not affect glucose tolerance in non-diabetics. Br J Clin Pharmacol 15:570–571CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Champy MF, Selloum M, Zeitler V et al (2008) Genetic background determines metabolic phenotypes in the mouse. Mamm Genome 19:318–331CrossRefPubMedGoogle Scholar
  42. 42.
    Brouwers B, de Faudeur G, Osipovich AB et al (2014) Impaired islet function in commonly used transgenic mouse lines due to human growth hormone minigene expression. Cell Metab 20:979–990CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Carboneau BA, Le TD, Dunn JC, Gannon M (2016) Unexpected effects of the MIP-CreER transgene and tamoxifen on β-cell growth in C57Bl6/J male mice. Physiological reports 4:e12863Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Kunhua Qin
    • 1
    • 2
  • Ning Zhang
    • 3
  • Zhao Zhang
    • 1
  • Michael Nipper
    • 2
  • Zhenxin Zhu
    • 2
  • Jake Leighton
    • 2
  • Kexin Xu
    • 1
  • Nicolas Musi
    • 3
  • Pei Wang
    • 2
  1. 1.Department of Molecular MedicineUniversity of Texas Health Science Centre at San AntonioSan AntonioUSA
  2. 2.Department of Cell Systems & AnatomyUniversity of Texas Health Science Centre at San AntonioSan AntonioUSA
  3. 3.Barshop Institute for Longevity and Aging StudiesUniversity of Texas Health Science Centre at San AntonioSan AntonioUSA

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