The somatic DNA methylation (DNAme) landscape is established early in development but remains highly dynamic within focal regions that overlap with gene regulatory elements. The significance of these dynamic changes, particularly in the central nervous system, remains unresolved. Here, we utilize a powerful human embryonic stem cell differentiation model for the generation of motor neurons (MNs) in combination with genetic mutations in the de novo DNAme machinery. We quantitatively dissect the role of DNAme in directing somatic cell fate with high-resolution genome-wide bisulfite-, bulk-, and single-cell-RNA sequencing. We find defects in neuralization and MN differentiation in DNMT3A knockouts (KO) that can be rescued by the targeting of DNAme to key developmental loci using catalytically inactive dCas9. We also find decreased dendritic arborization and altered electrophysiological properties in DNMT3A KO MNs. Our work provides a list of DNMT3A-regulated targets and a mechanistic link between de novo DNAme, cellular differentiation, and human MN function. Kiskinis and colleagues demonstrate that DNA methylation dynamics play a central role in the differentiation of human pluripotent stem cells toward highly specialized motor neurons. Through a combination of molecular and functional analysis they identify key transcriptional mediators of these effects and link DNA methylation to neuronal patterning and function.

Dissecting the Functional Consequences of De Novo DNA Methylation Dynamics in Human Motor Neuron Differentiation and Physiology / Ziller, Michael J.; Ortega, Juan A.; Quinlan, Katharina A.; Santos, David P.; Gu, Hongcang; Martin, Eric J.; Galonska, Christina; Pop, Ramona; Maidl, Susanne; DI PARDO, Alba; Huang, Mei; Meltzer, Herbert Y.; Gnirke, Andreas; Heckman, C. J.; Meissner, Alexander; Kiskinis, Evangelos. - In: CELL STEM CELL. - ISSN 1934-5909. - 22:4(2018), pp. 559-574.e9. [10.1016/j.stem.2018.02.012]

Dissecting the Functional Consequences of De Novo DNA Methylation Dynamics in Human Motor Neuron Differentiation and Physiology

DI PARDO, ALBA
Conceptualization
;
2018

Abstract

The somatic DNA methylation (DNAme) landscape is established early in development but remains highly dynamic within focal regions that overlap with gene regulatory elements. The significance of these dynamic changes, particularly in the central nervous system, remains unresolved. Here, we utilize a powerful human embryonic stem cell differentiation model for the generation of motor neurons (MNs) in combination with genetic mutations in the de novo DNAme machinery. We quantitatively dissect the role of DNAme in directing somatic cell fate with high-resolution genome-wide bisulfite-, bulk-, and single-cell-RNA sequencing. We find defects in neuralization and MN differentiation in DNMT3A knockouts (KO) that can be rescued by the targeting of DNAme to key developmental loci using catalytically inactive dCas9. We also find decreased dendritic arborization and altered electrophysiological properties in DNMT3A KO MNs. Our work provides a list of DNMT3A-regulated targets and a mechanistic link between de novo DNAme, cellular differentiation, and human MN function. Kiskinis and colleagues demonstrate that DNA methylation dynamics play a central role in the differentiation of human pluripotent stem cells toward highly specialized motor neurons. Through a combination of molecular and functional analysis they identify key transcriptional mediators of these effects and link DNA methylation to neuronal patterning and function.
2018
cell fate; DNA methylation; DNMT3A; epigenetics; ESCs; motor neurons; neurogenesis; spinal cord development; Molecular Medicine; Genetics; Cell Biology
01 Pubblicazione su rivista::01a Articolo in rivista
Dissecting the Functional Consequences of De Novo DNA Methylation Dynamics in Human Motor Neuron Differentiation and Physiology / Ziller, Michael J.; Ortega, Juan A.; Quinlan, Katharina A.; Santos, David P.; Gu, Hongcang; Martin, Eric J.; Galonska, Christina; Pop, Ramona; Maidl, Susanne; DI PARDO, Alba; Huang, Mei; Meltzer, Herbert Y.; Gnirke, Andreas; Heckman, C. J.; Meissner, Alexander; Kiskinis, Evangelos. - In: CELL STEM CELL. - ISSN 1934-5909. - 22:4(2018), pp. 559-574.e9. [10.1016/j.stem.2018.02.012]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1208178
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