Role of DNA Methylation and Hydroxymethylation in Epidermal Homeostasis

描述（由申请人提供）：这是一份申请K08临床科学家指导职业发展奖的申请，申请对象是加州大学旧金山分校的皮肤科医生Jeffrey Cheng博士。候选人的长期目标是成为表皮生物学和疾病表观遗传机制的主要独立研究者。这笔资助对于支持郑博士向知识独立和自己的研究项目方向过渡将是非常宝贵的。具体来说，K08奖将支持一个密集的5年培训计划，以发展强大的科学知识基础和计算生物学、上皮生物学和表观遗传学/表观基因组学方面的新技术技能，并提高拨款和科学写作技能。Joseph Costello博士将指导郑博士的科学培训和职业发展。科斯特洛博士是加州大学旧金山分校的神经外科教授，也是美国国立卫生研究院表观基因组图谱绘制中心的主任，他是研究脑癌及其发展的表观基因组学机制的领先研究员。擅长分析、整合和显示基于深度测序的表观基因组学数据的计算生物学家王婷博士和研究表皮分化和屏障功能的Theodora Mauro博士将共同指导Cheng博士。本研究旨在了解DNA甲基化和羟甲基化在表皮稳态中的作用。虽然DNA甲基化明显影响表皮稳态，但其在启动子外的基因调控作用尚不清楚。DNA羟甲基化在表皮中的作用尚未被研究。新出现的证据表明DNA甲基化和羟甲基化在增强子中起着重要作用。与此一致的是，初步的DNA甲基化数据显示，增殖性角化细胞中的大多数低甲基化区域与增强子重叠。为了探索DNA甲基化在增强子和DNA羟甲基化的表皮作用，Cheng博士将进行下一代基于测序的表观遗传分析分析，以识别增殖和分化角质形成细胞之间的差异甲基化和羟甲基化区域。接下来的实验将通过DNA甲基化或羟甲基化模式来验证角质形成细胞相关的候选增强子，并评估表皮DNA羟甲基化酶和5-羟甲基化缺失的功能后果。这些数据将成为帮助郑博士实现理解表皮疾病中表观遗传机制失调的长期目标的基础。

DESCRIPTION (provided by applicant): This is an application for a K08 Clinical Scientist Mentored Career Development Award for Dr. Jeffrey Cheng, a Dermatologist at the University of California, San Francisco. The candidate's long-term goal is to become a leading independent investigator of epigenetic mechanisms in epidermal biology and disease. This grant will be invaluable in supporting Dr. Cheng's transition towards intellectual independence and direction of his own research program. Specifically, this K08 award will support an intensive 5-year training plan to develop a strong scientific knowledge base and new technical skills in computational biology, epithelial biology, and epigenetics/epigenomics and to improve grant and scientific writing skills. Dr. Joseph Costello will mentor Dr. Cheng's scientific training and carer development. Dr. Costello, a Professor of Neurosurgery at UCSF and Director of a NIH Roadmap Epigenome Mapping Center, is a leading epigenomics researcher who studies epigenetic mechanisms in brain cancer and development. Dr. Ting Wang, a computational biologist with expertise in analyzing, integrating, and displaying deep sequencing-based epigenomics data, and Dr. Theodora Mauro, who studies epidermal differentiation and barrier function, will co-mentor Dr. Cheng. The proposed research seeks to understand the role of DNA methylation and hydroxymethylation in epidermal homeostasis. While DNA methylation clearly affects epidermal homeostasis, its gene regulatory role outside of promoters remains unclear. The role of DNA hydroxymethylation in the epidermis has not been studied. Emerging evidence suggests a significant role for DNA methylation and hydroxymethylation at enhancers. Consistent with this, preliminary DNA methylation data reveals that the majority of hypomethylated regions in proliferating keratinocytes overlap with enhancers. To explore the epidermal role of DNA methylation at enhancers and DNA hydroxymethylation, Dr. Cheng will perform next generation sequencing-based epigenetic profiling assays to identify differentially methylated and hydroxymethylated regions between proliferating and differentiating keratinocytes. This will be followed by experiments to validate keratinocyte- relevant candidate enhancers identified by their DNA methylation or hydroxymethylation patterns and assess the functional consequences of epidermal DNA hydroxymethylating enzyme and 5-hyroxymethylation depletion. These data will serve as a foundation to help achieve Dr. Cheng's long-term goal of understanding how epigenetic mechanisms are dysregulated in epidermal disease.