Role of DNA Methylation and Hydroxymethylation in Epidermal Homeostasis

描述(申请人提供)：这是为加州大学旧金山分校皮肤科医生杰弗里·程博士申请的K08临床科学家指导职业发展奖。这位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领先独立研究者。这笔拨款将对郑博士向知识独立的过渡和他自己的研究计划的方向提供无价的支持。具体地说，这项K08奖项将支持一项为期5年的强化培训计划，以发展计算生物学、上皮生物学和表观遗传学/表观基因组学方面强大的科学知识基础和新的技术技能，并提高赠款和科学写作技能。Joseph Costello博士将指导郑博士的科学培训和护理人员的发展。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NIH路线图表观基因组绘制中心的主任，他是领先的表观基因组学研究人员，研究脑癌和发育的表观遗传机制。在分析、整合和显示基于深度测序的表观基因组数据方面拥有专业知识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博士将共同指导程博士。这项拟议的研究试图了解DNA甲基化和羟甲基化在表皮动态平衡中的作用。虽然DNA甲基化明显影响表皮的动态平衡，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剂中起着重要作用。与此一致，初步的DNA甲基化数据显示，增殖角质形成细胞中的大多数低甲基化区域与增强剂重叠。为了探索表皮中DNA甲基化在增强剂和DNA羟甲基化中的作用，郑博士将进行基于下一代测序的表观遗传学分析，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。随后将进行实验，以验证通过DNA甲基化或羟甲基化模式确定的角质形成细胞相关候选增强剂，并评估表皮DNA羟甲基化酶和5-羟甲基化耗竭的功能后果。这些数据将有助于实现程博士的长期目标，即了解表观遗传机制在表皮疾病中是如何失调的。

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.