Cell and Tissue Research

, Volume 368, Issue 3, pp 563–578

Hyperglycemia impedes definitive endoderm differentiation of human embryonic stem cells by modulating histone methylation patterns

  • A. C. H. Chen
  • Y. L. Lee
  • S. W. Fong
  • C. C. Y. Wong
  • E. H. Y. Ng
  • W. S. B. Yeung
Regular Article

Abstract

Exposure to maternal diabetes during fetal growth is a risk factor for the development of type II diabetes (T2D) in later life. Discovery of the mechanisms involved in this association should provide valuable background for therapeutic treatments. Early embryogenesis involves epigenetic changes including histone modifications. The bivalent histone methylation marks H3K4me3 and H3K27me3 are important for regulating key developmental genes during early fetal pancreas specification. We hypothesized that maternal hyperglycemia disrupted early pancreas development through changes in histone bivalency. A human embryonic stem cell line (VAL3) was used as the cell model for studying the effects of hyperglycemia upon differentiation into definitive endoderm (DE), an early stage of the pancreatic lineage. Hyperglycemic conditions significantly down-regulated the expression levels of DE markers SOX17, FOXA2, CXCR4 and EOMES during differentiation. This was associated with retention of the repressive histone methylation mark H3K27me3 on their promoters under hyperglycemic conditions. The disruption of histone methylation patterns was observed as early as the mesendoderm stage, with Wnt/β-catenin signaling being suppressed during hyperglycemia. Treatment with Wnt/β-catenin signaling activator CHIR-99021 restored the expression levels and chromatin methylation status of DE markers, even in a hyperglycemic environment. The disruption of DE development was also found in mouse embryos at day 7.5 post coitum from diabetic mothers. Furthermore, disruption of DE differentiation in VAL3 cells led to subsequent impairment in pancreatic progenitor formation. Thus, early exposure to hyperglycemic conditions hinders DE development with a possible relationship to the later impairment of pancreas specification.

Keywords

hESCs Hyperglycemia Definitive endoderm Chromatin methylation Wnt/β-catenin signaling pathway 

Supplementary material

441_2017_2583_MOESM1_ESM.docx (1.6 mb)
ESM 1(DOCX 1641 kb)

References

  1. Aerts L, Vercruysse L, Van Assche FA (1997) The endocrine pancreas in virgin and pregnant offspring of diabetic pregnant rats. Diabetes Res Clin Pract 38:9–19CrossRefPubMedGoogle Scholar
  2. Amri K, Freund N, Vilar J, Merlet-Benichou C, Lelievre-Pegorier M (1999) Adverse effects of hyperglycemia on kidney development in rats: in vivo and in vitro studies. Diabetes 48:2240–2245CrossRefPubMedGoogle Scholar
  3. Anderson RM, Bosch JA, Goll MG, Hesselson D, Dong PD, Shin D, Chi NC, Shin CH, Schlegel A, Halpern M, Stainier DY (2009) Loss of Dnmt1 catalytic activity reveals multiple roles for DNA methylation during pancreas development and regeneration. Dev Biol 334:213–223CrossRefPubMedPubMedCentralGoogle Scholar
  4. Barker DJ (2004) The developmental origins of well-being. Philos Trans R Soc Lond B Biol Sci 359:1359–1366CrossRefPubMedPubMedCentralGoogle Scholar
  5. Boney CM, Verma A, Tucker R, Vohr BR (2005) Metabolic syndrome in childhood: association with birth weight, maternal obesity, and gestational diabetes mellitus. Pediatrics 115:e290–e296CrossRefPubMedGoogle Scholar
  6. Borowiak M, Maehr R, Chen S, Chen AE, Tang W, Fox JL, Schreiber SL, Melton DA (2009) Small molecules efficiently direct endodermal differentiation of mouse and human embryonic stem cells. Cell Stem Cell 4:348–358CrossRefPubMedPubMedCentralGoogle Scholar
  7. Bouchi R, Foo KS, Hua H, Tsuchiya K, Ohmura Y, Sandoval PR, Ratner LE, Egli D, Leibel RL, Accili D (2014) FOXO1 inhibition yields functional insulin-producing cells in human gut organoid cultures. Nat Commun 5:4242CrossRefPubMedPubMedCentralGoogle Scholar
  8. Bramswig NC, Everett LJ, Schug J, Dorrell C, Liu C, Luo Y, Streeter PR, Naji A, Grompe M, Kaestner KH (2013) Epigenomic plasticity enables human pancreatic alpha to beta cell reprogramming. J Clin Invest 123:1275–1284CrossRefPubMedPubMedCentralGoogle Scholar
  9. Brunner AL, Johnson DS, Kim SW, Valouev A, Reddy TE, Neff NF, Anton E, Medina C, Nguyen L, Chiao E, Oyolu CB, Schroth GP, Absher DM, Baker JC, Myers RM (2009) Distinct DNA methylation patterns characterize differentiated human embryonic stem cells and developing human fetal liver. Genome Res 19:1044–1056CrossRefPubMedPubMedCentralGoogle Scholar
  10. Cai J, Yu C, Liu Y, Chen S, Guo Y, Yong J, Lu W, Ding M, Deng H (2010) Generation of homogeneous PDX1(+) pancreatic progenitors from human ES cell-derived endoderm cells. J Mol Cell Biol 2:50–60CrossRefPubMedGoogle Scholar
  11. Cauza E, Hanusch-Enserer U, Strasser B, Ludvik B, Kostner K, Dunky A, Haber P (2005) Continuous glucose monitoring in diabetic long distance runners.Int J Sports Med 26:774–780Google Scholar
  12. Chen S, Borowiak M, Fox JL, Maehr R, Osafune K, Davidow L, Lam K, Peng LF, Schreiber SL, Rubin LL, Melton D (2009) A small molecule that directs differentiation of human ESCs into the pancreatic lineage. Nat Chem Biol 5:258–265CrossRefPubMedGoogle Scholar
  13. Christman JK (2002) 5-Azacytidine and 5-aza-2′-deoxycytidine as inhibitors of DNA methylation: mechanistic studies and their implications for cancer therapy. Oncogene 21:5483–5495CrossRefPubMedGoogle Scholar
  14. Costello I, Pimeisl IM, Drager S, Bikoff EK, Robertson EJ, Arnold SJ (2011) The T-box transcription factor Eomesodermin acts upstream of Mesp1 to specify cardiac mesoderm during mouse gastrulation. Nat Cell Biol 13:1084–1091CrossRefPubMedPubMedCentralGoogle Scholar
  15. D’Amour KA, Agulnick AD, Eliazer S, Kelly OG, Kroon E, Baetge EE (2005) Efficient differentiation of human embryonic stem cells to definitive endoderm. Nat Biotechnol 23:1534–1541CrossRefPubMedGoogle Scholar
  16. Dhara SK, Hasneen K, Machacek DW, Boyd NL, Rao RR, Stice SL (2008) Human neural progenitor cells derived from embryonic stem cells in feeder-free cultures. Differ Res Biol Divers 76:454–464CrossRefGoogle Scholar
  17. Dhawan S, Georgia S, Tschen SI, Fan G, Bhushan A (2011) Pancreatic beta cell identity is maintained by DNA methylation-mediated repression of Arx. Dev Cell 20:419–429CrossRefPubMedPubMedCentralGoogle Scholar
  18. Ding GL, Wang FF, Shu J, Tian S, Jiang Y, Zhang D, Wang N, Luo Q, Zhang Y, Jin F, Leung PC, Sheng JZ, Huang HF (2012) Transgenerational glucose intolerance with Igf2/H19 epigenetic alterations in mouse islet induced by intrauterine hyperglycemia. Diabetes 61:1133–1142CrossRefPubMedPubMedCentralGoogle Scholar
  19. Feit-Leichman RA, Kinouchi R, Takeda M, Fan Z, Mohr S, Kern TS, Chen DF (2005) Vascular damage in a mouse model of diabetic retinopathy: relation to neuronal and glial changes. Invest Ophthalmol Vis Sci 46:4281–4287CrossRefPubMedGoogle Scholar
  20. Gadue P, Huber TL, Paddison PJ, Keller GM (2006) Wnt and TGF-beta signaling are required for the induction of an in vitro model of primitive streak formation using embryonic stem cells. Proc Natl Acad Sci U S A 103:16806–16811CrossRefPubMedPubMedCentralGoogle Scholar
  21. Gluckman PD, Hanson MA (2007) Developmental plasticity and human disease: research directions. J Intern Med 261:461–471CrossRefPubMedGoogle Scholar
  22. Jackson SA, Schiesser J, Stanley EG, Elefanty AG (2010) Differentiating embryonic stem cells pass through “temporal windows” that mark responsiveness to exogenous and paracrine mesendoderm inducing signals. PLoS One 5:e10706CrossRefPubMedPubMedCentralGoogle Scholar
  23. Jiang W, Wang J, Zhang Y (2013) Histone H3K27me3 demethylases KDM6A and KDM6B modulate definitive endoderm differentiation from human ESCs by regulating WNT signaling pathway. Cell Res 23:122–130CrossRefPubMedGoogle Scholar
  24. Jufvas A, Sjodin S, Lundqvist K, Amin R, Vener AV, Stralfors P (2013) Global differences in specific histone H3 methylation are associated with overweight and type 2 diabetes. Clin Epigenet 5:15CrossRefGoogle Scholar
  25. Kanai-Azuma M, Kanai Y, Gad JM, Tajima Y, Taya C, Kurohmaru M, Sanai Y, Yonekawa H, Yazaki K, Tam PP, Hayashi Y (2002) Depletion of definitive gut endoderm in Sox17-null mutant mice. Development 129:2367–2379PubMedGoogle Scholar
  26. Kelstrup L, Hjort L, Houshmand-Oeregaard A, Clausen TD, Hansen NS, Broholm C, Borch-Johnsen L, Mathiesen ER, Vaag AA, Damm P (2016) Gene expression and DNA methylation of PPARGC1A in muscle and adipose tissue from adult offspring of women with diabetes in pregnancy. Diabetes 65:2900–2910CrossRefPubMedGoogle Scholar
  27. Kim MH, Hong SH, Lee MK (2013) Insulin receptor-overexpressing beta-cells ameliorate hyperglycemia in diabetic rats through Wnt signaling activation. PLoS One 8:e67802CrossRefPubMedPubMedCentralGoogle Scholar
  28. Kubo A, Shinozaki K, Shannon JM, Kouskoff V, Kennedy M, Woo S, Fehling HJ, Keller G (2004) Development of definitive endoderm from embryonic stem cells in culture. Development 131:1651–1662CrossRefPubMedGoogle Scholar
  29. Kumar M, Melton D (2003) Pancreas specification: a budding question. Curr Opin Genet Dev 13:401–407CrossRefPubMedGoogle Scholar
  30. Kwok CT, Leung MH, Qin J, Qin Y, Wang J, Lee YL, Yao KM (2016) The Forkhead box transcription factor FOXM1 is required for the maintenance of cell proliferation and protection against oxidative stress in human embryonic stem cells. Stem Cell Res 16:651–661CrossRefPubMedGoogle Scholar
  31. Li Z, Nie F, Wang S, Li L (2011) Histone H4 Lys 20 monomethylation by histone methylase SET8 mediates Wnt target gene activation. Proc Natl Acad Sci U S A 108:3116–3123CrossRefPubMedPubMedCentralGoogle Scholar
  32. Metcalfe C, Bienz M (2011) Inhibition of GSK3 by Wnt signalling—two contrasting models. J Cell Sci 124:3537–3544CrossRefPubMedGoogle Scholar
  33. Miranda TB, Cortez CC, Yoo CB, Liang G, Abe M, Kelly TK, Marquez VE, Jones PA (2009) DZNep is a global histone methylation inhibitor that reactivates developmental genes not silenced by DNA methylation. Mol Cancer Ther 8:1579–1588CrossRefPubMedPubMedCentralGoogle Scholar
  34. Mochizuki H, Ohnuki Y, Kurosawa H (2011) Effect of glucose concentration during embryoid body (EB) formation from mouse embryonic stem cells on EB growth and cell differentiation. J Biosci Bioeng 111:92–97CrossRefPubMedGoogle Scholar
  35. Ninomiya H, Mizuno K, Terada R, Miura T, Ohnuma K, Takahashi S, Asashima M, Michiue T (2015) Improved efficiency of definitive endoderm induction from human induced pluripotent stem cells in feeder and serum-free culture system. In Vitro Cell Dev Biol Animal 51:1–8CrossRefGoogle Scholar
  36. Pagliuca FW, Millman JR, Gurtler M, Segel M, Van Dervort A, Ryu JH, Peterson QP, Greiner D, Melton DA (2014) Generation of functional human pancreatic beta cells in vitro. Cell 159:428–439CrossRefPubMedPubMedCentralGoogle Scholar
  37. Pasini D, Bracken AP, Jensen MR, Lazzerini Denchi E, Helin K (2004) Suz12 is essential for mouse development and for EZH2 histone methyltransferase activity. EMBO J 23:4061–4071CrossRefPubMedPubMedCentralGoogle Scholar
  38. Rada-Iglesias A, Bajpai R, Swigut T, Brugmann SA, Flynn RA, Wysocka J (2011) A unique chromatin signature uncovers early developmental enhancers in humans. Nature 470:279–283CrossRefPubMedGoogle Scholar
  39. Rezania A, Bruin JE, Arora P, Rubin A, Batushansky I, Asadi A, O’Dwyer S, Quiskamp N, Mojibian M, Albrecht T, Yang YH, Johnson JD, Kieffer TJ (2014) Reversal of diabetes with insulin-producing cells derived in vitro from human pluripotent stem cells. Nat Biotechnol 32:1121–1133CrossRefPubMedGoogle Scholar
  40. Ruiz-Munoz LM, Vidal-Vanaclocha F, Lampreabe I (1997) Enalaprilat inhibits hydrogen peroxide production by murine mesangial cells exposed to high glucose concentrations. Nephrol Dial Transplant 12:456–464CrossRefPubMedGoogle Scholar
  41. Salbaum JM, Kappen C (2012) Responses of the embryonic epigenome to maternal diabetes. Birth Defects Res Part A Clin Mol Teratol 94:770–781CrossRefPubMedGoogle Scholar
  42. Shea K, Geijsen N (2007) Dissection of 6.5 dpc mouse embryos. J Vis Exp 2007:160Google Scholar
  43. Silverman BL, Metzger BE, Cho NH, Loeb CA (1995) Impaired glucose tolerance in adolescent offspring of diabetic mothers. Relationship to fetal hyperinsulinism. Diabetes Care 18:611–617CrossRefPubMedGoogle Scholar
  44. Soria B, Roche E, Berna G, Leon-Quinto T, Reig JA, Martin F (2000) Insulin-secreting cells derived from embryonic stem cells normalize glycemia in streptozotocin-induced diabetic mice. Diabetes 49:157–162CrossRefPubMedGoogle Scholar
  45. Tada S, Era T, Furusawa C, Sakurai H, Nishikawa S, Kinoshita M, Nakao K, Chiba T, Nishikawa S (2005) Characterization of mesendoderm: a diverging point of the definitive endoderm and mesoderm in embryonic stem cell differentiation culture. Development 132:4363–4374CrossRefPubMedGoogle Scholar
  46. Takei S, Ichikawa H, Johkura K, Mogi A, No H, Yoshie S, Tomotsune D, Sasaki K (2009) Bone morphogenetic protein-4 promotes induction of cardiomyocytes from human embryonic stem cells in serum-based embryoid body development. Am J Physiol Heart Circ Physiol 296:H1793–H1803CrossRefPubMedGoogle Scholar
  47. Teo AK, Arnold SJ, Trotter MW, Brown S, Ang LT, Chng Z, Robertson EJ, Dunn NR, Vallier L (2011) Pluripotency factors regulate definitive endoderm specification through eomesodermin. Genes Dev 25:238–250CrossRefPubMedPubMedCentralGoogle Scholar
  48. Valbuena D, Galan A, Sanchez E, Poo ME, Gomez E, Sanchez-Luengo S, Melguizo D, Garcia A, Ruiz V, Moreno R, Pellicer A, Simon C (2006) Derivation and characterization of three new Spanish human embryonic stem cell lines (VAL −3 -4 -5) on human feeder and in serum-free conditions. Reprod Biomed Online 13:875–886CrossRefPubMedGoogle Scholar
  49. Ventura-Sobrevilla J, Boone-Villa VD, Aguilar CN, Roman-Ramos R, Vega-Avila E, Campos-Sepulveda E, Alarcon-Aguilar F (2011) Effect of varying dose and administration of streptozotocin on blood sugar in male CD1 mice. Proc West Pharmacol Soc 54:5–9PubMedGoogle Scholar
  50. Wang P, Rodriguez RT, Wang J, Ghodasara A, Kim SK (2011) Targeting SOX17 in human embryonic stem cells creates unique strategies for isolating and analyzing developing endoderm. Cell Stem Cell 8:335–346CrossRefPubMedPubMedCentralGoogle Scholar
  51. Wohrle S, Wallmen B, Hecht A (2007) Differential control of Wnt target genes involves epigenetic mechanisms and selective promoter occupancy by T-cell factors. Mol Cell Biol 27:8164–8177CrossRefPubMedPubMedCentralGoogle Scholar
  52. Wong AP, Chin S, Xia S, Garner J, Bear CE, Rossant J (2015) Efficient generation of functional CFTR-expressing airway epithelial cells from human pluripotent stem cells. Nat Protoc 10:363–381CrossRefPubMedGoogle Scholar
  53. Wong TY, Phillips AO, Witowski J, Topley N (2003) Glucose-mediated induction of TGF-beta 1 and MCP-1 in mesothelial cells in vitro is osmolality and polyol pathway dependent. Kidney Int 63:1404–1416CrossRefPubMedGoogle Scholar
  54. Xie R, Everett LJ, Lim HW, Patel NA, Schug J, Kroon E, Kelly OG, Wang A, D’Amour KA, Robins AJ, Won KJ, Kaestner KH, Sander M (2013) Dynamic chromatin remodeling mediated by polycomb proteins orchestrates pancreatic differentiation of human embryonic stem cells. Cell Stem Cell 12:224–237CrossRefPubMedPubMedCentralGoogle Scholar
  55. Yang BT, Dayeh TA, Volkov PA, Kirkpatrick CL, Malmgren S, Jing X, Renstrom E, Wollheim CB, Nitert MD, Ling C (2012) Increased DNA methylation and decreased expression of PDX-1 in pancreatic islets from patients with type 2 diabetes. Mol Endocrinol 26:1203–1212CrossRefPubMedGoogle Scholar
  56. Ying L, Mills JA, French DL, Gadue P (2015) OCT4 coordinates with WNT signaling to pre-pattern chromatin at the SOX17 locus during human ES cell differentiation into definitive endoderm. Stem Cell Rep 5:490–498CrossRefGoogle Scholar
  57. Zhang F, Hong S, Stone V, Smith PJ (2007) Expression of cannabinoid CB1 receptors in models of diabetic neuropathy. J Pharmacol Exp Ther 323:508–515CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • A. C. H. Chen
    • 1
  • Y. L. Lee
    • 1
    • 2
    • 3
    • 4
  • S. W. Fong
    • 1
  • C. C. Y. Wong
    • 1
  • E. H. Y. Ng
    • 1
    • 2
    • 3
  • W. S. B. Yeung
    • 1
    • 2
    • 3
  1. 1.Department of Obstetrics and Gynaecology, LKS Faculty of MedicineThe University of Hong KongHong KongPeople’s Republic of China
  2. 2.Shenzhen Key Laboratory of Fertility Regulation, The University of Hong Kong Shenzhen HospitalThe University of Hong KongShenzhenPeople’s Republic of China
  3. 3.Center for Reproduction, Development and Growth, LKS Faculty of MedicineThe University of Hong KongHong KongPeople’s Republic of China
  4. 4.Department of Obstetrics and Gynaecology, LKS Faculty of MedicineThe University of Hong KongHong KongPeople’s Republic of China

Personalised recommendations