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