DNA methylation in orofacial clefting

了解可塑性表观遗传机制在出生缺陷中的作用是预防的直接途径 战略布局唇腭裂是人类出生时最常见的结构性畸形之一 缺陷，影响800个新生儿中的1个，并造成严重的个人，家庭和社会负担。预防 OFC的策略是难以捉摸的，因为我们目前对致病因素的理解是不够的。 流行病学和传统遗传学研究表明，OFC是病因复杂的结果， 这是多因素遗传和环境影响的结果。表观遗传机制是一个令人兴奋的新的 重点是理解OFC的起源，因为它们介导了环境影响对环境的影响。 基因组在敏感的胚胎时期。我们的建议特别关注DNA甲基化， 这种表观遗传机制是环境敏感的，并且是预防和治疗的实际目标 战略布局虽然有多方面的证据表明，DNA甲基化在口腔颌面部的生物学作用， 发展不清楚。我们已经建立了新颖的模型，并生成了使我们保持平衡的概念的关键证明 揭示DNA甲基化如何调节口面形态发生，并确定DNA 甲基化在调节OFC易感性中起作用。在这个项目中，整合的全基因组甲基化和 批量和单细胞转录组的方法将被应用于定义的分子和细胞机制， 颅神经嵴DNA甲基化破坏导致的OFC发病机制DNA的作用 多因素OFC易感性中的甲基化将通过整合多种环境和 遗传（Wnt9b KO）和化学（Shh拮抗剂）小鼠模型的DNA甲基化的饮食调节剂 不完全渗透的OFC。最后，表观基因组编辑将被应用于评估 OFC相关甲基化改变基因表达和颅神经嵴生物学。完成 拟议中的研究将带来对DNA甲基化如何调节颅神经嵴的基本见解 生物学和口面形态发生。通过定义环境和饮食介导的甲基化， 转录组反应改变OFC的易感性，这些研究也将提供必要的基础， 识别调节DNA甲基化并导致OFC风险的环境影响。 开展这一调查将推进我们制定预防战略的长期目标， 病因复杂的出生缺陷，通过识别有罪的环境影响，并确定其 行动机制。

Understanding the role of malleable epigenetic mechanisms in birth defects is a direct path to prevention strategies. Orofacial clefts (OFCs) of the lip and palate are among the most common human structural birth defects, affecting 1 in 800 newborns, and pose serious individual, familial, and societal burdens. Prevention strategies for OFCs are elusive because our current understanding of causative factors is inadequate. Epidemiologic and traditional genetic studies have shown that OFCs are etiologically complex outcomes that result from multifactorial genetic and environmental influences. Epigenetic mechanisms are an exciting new focus in understanding the genesis of OFCs because they mediate the effect of environmental influences on the genome during sensitive embryonic periods. Our proposal specifically focuses on DNA methylation because this epigenetic mechanism is environmentally sensitive and a practical target of prevention and therapeutic strategies. While implicated by multiple lines of evidence, the biological role of DNA methylation in orofacial development is unclear. We have established novel models and generated key proofs of concepts that poise us to uncover how DNA methylation regulates orofacial morphogenesis and to define the role that DNA methylation plays in modulating OFC susceptibility. In this project, integrated genome-wide methylation and bulk and single-cell transcriptome approaches will be applied to define molecular and cellular mechanisms of OFC pathogenesis resulting from disrupted DNA methylation in the cranial neural crest. The role of DNA methylation in multifactorial OFC susceptibility will then be defined by integrating multiple environmental and dietary modulators of DNA methylation to genetic (Wnt9b KO) and chemical (Shh antagonist) mouse models of incompletely penetrant OFCs. Finally, epigenome editing will be applied to evaluate the functional impact of OFC-associated methylation changes on gene expression and cranial neural crest biology. Completion of the proposed studies will bring fundamental insight into how DNA methylation regulates cranial neural crest biology and orofacial morphogenesis. By defining environmental- and dietary-mediated methylome- transcriptome responses that alter OFC susceptibility, these studies will also provide a necessary foundation for identification of environmental influences that modulate DNA methylation and contribute to OFC risk. Pursuing this line of investigation will advance our long-term goal of developing prevention strategies for etiologically complex birth defects by identifying culpable environmental influences and defining their mechanisms of action.