Tet-mediated imprinting erasure in H19 locus following reprogramming of spermatogonial stem cells to induced pluripotent stem cells.
Selective methylation of CpG islands at imprinting control regions (ICR) determines the monoparental expression of a subset of genes. Currently, it is unclear whether artificial reprogramming induced by the expression of Yamanaka factors disrupts these marks and whether cell type of origin affects the dynamics of reprogramming. In this study, spermatogonial stem cells (SSC) that harbor paternalized imprinting marks, and fibroblasts were reprogrammed to iPSC (SSCiPSC and fiPSC). The SSCiPSC were able to form teratomas and generated chimeras with a higher skin chimerism than those derived from fiPSC. RNA-seq revealed extensive reprogramming at the transcriptional level with 8124 genes differentially expressed between SSC and SSCiPSC and only 490 between SSCiPSC and fiPSC. Likewise, reprogramming of SSC affected 26 of 41 imprinting gene clusters known in the mouse genome. A closer look at H19 ICR revealed complete erasure in SSCiPSC in contrast to fiPSC. Imprinting erasure in SSCiPSC was maintained even after in vivo differentiation into teratomas. Reprogramming of SSC from Tet1 and Tet2 double knockout mice however lacked demethylation of H19 ICR. These results suggest that imprinting erasure during reprogramming depends on the epigenetic landscape of the precursor cell and is mediated by TETs at the H19 locus.
Department of Animal and Avian Sciences, University of Maryland, MD, USA.|Animal Bioscience and Biotechnology Laboratory, USDA-ARS, Beltsville, MD, USA.|Departamento de Reproducción Animal, INIA, Madrid, Spain.
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Bermejo-Álvarez
Departamento de Reproducción Animal, INIA, Madrid, Spain.
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P
Ramos-Ibeas
Department of Animal and Avian Sciences, University of Maryland, MD, USA.|Animal Bioscience and Biotechnology Laboratory, USDA-ARS, Beltsville, MD, USA.
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KE
Park
Animal Bioscience and Biotechnology Laboratory, USDA-ARS, Beltsville, MD, USA.
A P
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Powell
Department of Animal and Avian Sciences, University of Maryland, MD, USA.
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Vansandt
Animal Improvement Program Laboratory, USDA-ARS, Beltsville, MD, USA.
Bickhart
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Derek
Departamento de Reproducción Animal, INIA, Madrid, Spain.
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MA
Ramirez
Departamento de Reproducción Animal, INIA, Madrid, Spain.
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Gutiérrez-Adán
Department of Animal and Avian Sciences, University of Maryland, MD, USA.|Animal Bioscience and Biotechnology Laboratory, USDA-ARS, Beltsville, MD, USA.
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BP
Telugu
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D065150:Cellular Reprogramming; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism*; D049308:Dioxygenases; D020869:Gene Expression Profiling; D056426:Genetic Loci; D018392:Genomic Imprinting; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D013093:Spermatogonia / Q000166:cytology*
D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; C540839:TET1 protein, mouse; D049308:Dioxygenases; C551914:Tet2 protein, mouse