Hyperglycemia impedes definitive endoderm differentiation of human embryonic stem cells by modulating histone methylation patterns
- 411 Downloads
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.
KeywordshESCs Hyperglycemia Definitive endoderm Chromatin methylation Wnt/β-catenin signaling pathway
We thank the Príncipe Felipe Research Center of Valencia for providing the hES cell line, VAL3. We are also grateful to the Department of Biochemistry, The University of Hong Kong for providing the mES cell line, L4. We additionally thank the Faculty Core Facility, The University of Hong Kong for assistance with flow cytometry and confocal microscopic analyses. This project is supported by the Small Project Funding of the University of Hong Kong Committee on Research and Conference Grants.
C.H.C and Y.L.L. designed and performed the experiments, interpreted the data and wrote the manuscript. S.W.F. and C.Y.W. assisted with the experiments. Y.H.N. and S.B.Y. supervised the project and assisted with the experimental design and with writing the manuscript.
Compliance with ethical standards
Conflict of interests
All authors state that they have no competing financial interests.
- 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
- 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
- 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
- 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
- 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
- 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
- Shea K, Geijsen N (2007) Dissection of 6.5 dpc mouse embryos. J Vis Exp 2007:160Google Scholar
- 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
- 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
- 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