TitleDisruption of DNA methylation-mediated cranial neural crest proliferation and differentiation causes orofacial clefts in mice.
Publication TypeJournal Article
Year of Publication2024
AuthorsUlschmid CM, Sun MR, Jabbarpour CR, Steward AC, Rivera-González KS, Cao J, Martin AA, Barnes M, Wicklund L, Madrid A, Papale LA, Joseph DB, Vezina CM, Alisch RS, Lipinski RJ
JournalProc Natl Acad Sci U S A
Date Published2024 Jan 16
KeywordsAnimals, Cell Proliferation, Cleft Lip, Cleft Palate, DNA Methylation, DNA Modification Methylases, Mice, Neural Crest

Orofacial clefts of the lip and palate are widely recognized to result from complex gene-environment interactions, but inadequate understanding of environmental risk factors has stymied development of prevention strategies. We interrogated the role of DNA methylation, an environmentally malleable epigenetic mechanism, in orofacial development. Expression of the key DNA methyltransferase enzyme DNMT1 was detected throughout palate morphogenesis in the epithelium and underlying cranial neural crest cell (cNCC) mesenchyme, a highly proliferative multipotent stem cell population that forms orofacial connective tissue. Genetic and pharmacologic manipulations of DNMT activity were then applied to define the tissue- and timing-dependent requirement of DNA methylation in orofacial development. cNCC-specific inactivation targeting initial palate outgrowth resulted in OFCs, while later targeting during palatal shelf elevation and elongation did not. Conditional deletion reduced cNCC proliferation and subsequent differentiation trajectory, resulting in attenuated outgrowth of the palatal shelves and altered development of cNCC-derived skeletal elements. Finally, we found that the cellular mechanisms of cleft pathogenesis observed in vivo can be recapitulated by pharmacologically reducing DNA methylation in multipotent cNCCs cultured in vitro. These findings demonstrate that DNA methylation is a crucial epigenetic regulator of cNCC biology, define a critical period of development in which its disruption directly causes OFCs, and provide opportunities to identify environmental influences that contribute to OFC risk.

Alternate JournalProc Natl Acad Sci U S A
PubMed ID38194455
PubMed Central IDPMC10801837
Grant ListR03 DE027162 / DE / NIDCR NIH HHS / United States
U01 DK110807 / DK / NIDDK NIH HHS / United States
R56 DE030917 / DE / NIDCR NIH HHS / United States
T32 ES007015 / ES / NIEHS NIH HHS / United States
R01 DE032710 / DE / NIDCR NIH HHS / United States
R01 DK099328 / DK / NIDDK NIH HHS / United States