TY - JOUR
T1 - DNMT3B supports meso-endoderm differentiation from mouse embryonic stem cells
AU - Lauria, Andrea
AU - Meng, Guohua
AU - Proserpio, Valentina
AU - Rapelli, Stefania
AU - Maldotti, Mara
AU - Polignano, Isabelle Laurence
AU - Anselmi, Francesca
AU - Incarnato, Danny
AU - Krepelova, Anna
AU - Donna, Daniela
AU - Levra Levron, Chiara
AU - Donati, Giacomo
AU - Molineris, Ivan
AU - Neri, Francesco
AU - Oliviero, Salvatore
N1 - Funding Information:
We thank Graziano Martello (University of Padova) for the critical reading of the manuscript. V.P. was supported by Fondazione Umberto Veronesi (FUV). S.O. was supported by the Associazione Italiana per la Ricerca sul Cancro (AIRC) IG 2017 Id. 20240, and IG IG 2022 ID 27155, PRIN 2018, and IIGM institutional funds. GD was supported by AIRC MFAG 1018 Id 21640.
Funding Information:
We thank Graziano Martello (University of Padova) for the critical reading of the manuscript. V.P. was supported by Fondazione Umberto Veronesi (FUV). S.O. was supported by the Associazione Italiana per la Ricerca sul Cancro (AIRC) IG 2017 Id. 20240, and IG IG 2022 ID 27155, PRIN 2018, and IIGM institutional funds. GD was supported by AIRC MFAG 1018 Id 21640.
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/1
Y1 - 2023/1
N2 - The correct establishment of DNA methylation patterns during mouse early development is essential for cell fate specification. However, the molecular targets as well as the mechanisms that determine the specificity of the de novo methylation machinery during differentiation are not completely elucidated. Here we show that the DNMT3B-dependent DNA methylation of key developmental regulatory regions at epiblast-like cells (EpiLCs) provides an epigenetic priming that ensures flawless commitment at later stages. Using in vitro stem cell differentiation and loss of function experiments combined with high-throughput genome-wide bisulfite-, bulk-, and single cell RNA-sequencing we dissected the specific role of DNMT3B in cell fate. We identify DNMT3B-dependent regulatory elements on the genome which, in Dnmt3b knockout (3BKO), impair the differentiation into meso-endodermal (ME) progenitors and redirect EpiLCs towards the neuro-ectodermal lineages. Moreover, ectopic expression of DNMT3B in 3BKO re-establishes the DNA methylation of the master regulator Sox2 super-enhancer, downmodulates its expression, and restores the expression of ME markers. Taken together, our data reveal that DNMT3B-dependent methylation at the epiblast stage is essential for the priming of the meso-endodermal lineages and provide functional characterization of the de novo DNMTs during EpiLCs lineage determination.
AB - The correct establishment of DNA methylation patterns during mouse early development is essential for cell fate specification. However, the molecular targets as well as the mechanisms that determine the specificity of the de novo methylation machinery during differentiation are not completely elucidated. Here we show that the DNMT3B-dependent DNA methylation of key developmental regulatory regions at epiblast-like cells (EpiLCs) provides an epigenetic priming that ensures flawless commitment at later stages. Using in vitro stem cell differentiation and loss of function experiments combined with high-throughput genome-wide bisulfite-, bulk-, and single cell RNA-sequencing we dissected the specific role of DNMT3B in cell fate. We identify DNMT3B-dependent regulatory elements on the genome which, in Dnmt3b knockout (3BKO), impair the differentiation into meso-endodermal (ME) progenitors and redirect EpiLCs towards the neuro-ectodermal lineages. Moreover, ectopic expression of DNMT3B in 3BKO re-establishes the DNA methylation of the master regulator Sox2 super-enhancer, downmodulates its expression, and restores the expression of ME markers. Taken together, our data reveal that DNMT3B-dependent methylation at the epiblast stage is essential for the priming of the meso-endodermal lineages and provide functional characterization of the de novo DNMTs during EpiLCs lineage determination.
UR - http://www.scopus.com/inward/record.url?scp=85146771241&partnerID=8YFLogxK
U2 - 10.1038/s41467-023-35938-x
DO - 10.1038/s41467-023-35938-x
M3 - Article
C2 - 36690616
AN - SCOPUS:85146771241
SN - 2041-1723
VL - 14
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 367
ER -