Deletion of the CTG Expansion in Myotonic Dystrophy Type 1 Reverses DMPK Aberrant Methylation in Human Embryonic Stem Cells but not Affected Myoblasts
Deletion of the CTG Expansion in Myotonic Dystrophy Type 1 Reverses DMPK Aberrant Methylation in Human Embryonic Stem Cells but not Affected Myoblasts
ABSTRACT Myotonic dystrophy type 1 (DM1) results from a CTG repeat expansion in the 3’-UTR of DMPK. When the repeat extensively expands, this results in DMPK aberrant methylation, reduction in SIX5 transcription and the development of the congenital form of the disease. To explore whether hypermethylation could be reversed in DM1 embryonic stem cells (hESCs) and patient myoblasts, we monitored methylation levels following removal of the expanded repeat by CRISPR/Cas9-mediated editing. Excision of the repeat in undifferentiated hESCs (CTG2000) resets the locus by abolishing abnormal methylation and H3K9me3 enrichment, and rescues SIX5 transcription. In contrast, in affected myoblasts methylation levels remain unchanged following deletion of a large expansion (CTG2600). Altogether, this provides evidence for a transition from a reversible to an irreversible heterochromatin state by the DM1 mutation upon cell differentiation. These findings should be taken into account when considering gene correction in congenital DM1 and potentially other epigenetically regulated disorders.
Epsztejn-Litman Silvina、Bnaya Ester、Handal Tayma、Zahdeh Fouad、van den Broek Walther J.A.A.、Eiges Rachel、Diab Manar Abu、Yanovsky-Dagan Shira、Wansink Derick G.
Stem Cell Research Laboratory, Shaare Zedek Medical CenterStem Cell Research Laboratory, Shaare Zedek Medical Center||The Hebrew University School of MedicineStem Cell Research Laboratory, Shaare Zedek Medical CenterMedical Genetics Institute, Shaare Zedek Medical CenterDepartment of Cell Biology, Radboud Institute for Molecular Life Sciences (RIMLS), Radboud University Medical CenterStem Cell Research Laboratory, Shaare Zedek Medical Center||The Hebrew University School of MedicineStem Cell Research Laboratory, Shaare Zedek Medical Center||The Hebrew University School of MedicineStem Cell Research Laboratory, Shaare Zedek Medical Center||The Hebrew University School of MedicineDepartment of Cell Biology, Radboud Institute for Molecular Life Sciences (RIMLS), Radboud University Medical Center
基础医学遗传学分子生物学
Epsztejn-Litman Silvina,Bnaya Ester,Handal Tayma,Zahdeh Fouad,van den Broek Walther J.A.A.,Eiges Rachel,Diab Manar Abu,Yanovsky-Dagan Shira,Wansink Derick G..Deletion of the CTG Expansion in Myotonic Dystrophy Type 1 Reverses DMPK Aberrant Methylation in Human Embryonic Stem Cells but not Affected Myoblasts[EB/OL].(2025-03-28)[2025-05-07].https://www.biorxiv.org/content/10.1101/631457.点此复制
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