Functional analysis of methylation in neural crest migration

神经嵴迁移甲基化的功能分析

• 批准号: 8049711
• 负责人: Katie L. Vermillion
• 金额: $ 2.8万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-21 至 2013-04-20
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8049711
• 关键词: Ablation; Biological; Biological Assay; Cell Nucleus; Cells; Cleaved cell; Congenital Abnormality; Congenital Heart Defects; Cytoplasm; DNA Methylation; Data; Defect; Development; Dominant-Negative Mutation; Electroporation; Embryo; Ensure; Epitopes; Face; Folate; Folic Acid Deficiency; Goals; Health; Heart; Histones; Immunofluorescence Immunologic; In Situ Hybridization; Incidence; Lead; Malignant Neoplasms; Mediating; Methylation; Methyltransferase; Molecular; Neoplasm Metastasis; Nervous System Part; Neural Crest; Neural Crest Cell; Nuclear; Oligonucleotides; Pattern; Peripheral Nervous System; Process; Property; Proteins; Reaction; Research; Role; Site; Skeleton; Skin; Specificity; Supplementation; Testing; Work; base; cell motility; cofactor; craniofacial; dietary supplements; insight; knock-down; melanocyte; migration; migratory population; multipotent cell; novel; protective effect; research study; response; slug; tumor progression

DESCRIPTION (provided by applicant): Neural crest cells are a population of migratory, multipotent cells that differentiate into many diverse vertebrate lineages, including the craniofacial skeleton, peripheral nervous system, and melanocytes. Deficiencies in neural crest development lead to a variety of birth defects, including craniofacial clefts and some congenital heart defects. Folate supplements reduce the incidence of these birth defects, but the mechanism of this effect is unknown. One possibility is that folates rescue methylation. Although methylation has never been implicated in neural crest development, my preliminary data show that methylation is required for neural crest migration. Based on the expression of the methyltransferase Ezh2 in the cytoplasm of migratory neural crest cells, I postulate that non-histone protein methylation controls the migratory properties of neural crest cells. This proposal seeks to test this hypothesis and to identify and characterize the methyltransferases that mediate methylation in the neural crest through three specific aims: (1) The spatial and temporal localization of three candidate methyltransferases will be determined by immunofluorescence. (2) Antisense morpholino oligonucleotide-mediated protein ablation will be used to assay the requirement for each candidate methyltransferases during neural crest development. (3) The specific requirement for the methyltransferase Ezh2 in the cytoplasm and the nucleus will be assessed using dominant negative constructs. Once the requirement for particular methyltransferases is established in the neural crest, we can begin to look at the effects of folate-deficiency on methylation, and elucidate the mechanism by which folate supplementation reduces birth defects. This work will also provide insight into the molecular mechanisms of cancer metastasis. PUBLIC HEALTH RELEVANCE: Neural crest cells are cells that move through the developing embryo to form parts of the nervous system, face, heart and skin. The goal of this research is to determine how neural crest cells are able to move throughout the body. This process is similar to cancer and will help us to understand cancer progression, and will also provide us with a better understanding of how defects in neural crest cell movement can lead to birth defects.

描述（由申请人提供）：神经rest细胞是迁移的多能细胞群，它们分化为许多多种脊椎动物谱系，包括颅面骨骼，周围神经系统和黑素细胞。神经波峰发育的缺陷导致各种出生缺陷，包括颅面裂缝和一些先天性心脏缺陷。叶酸补充剂减少了这些先天缺陷的发生率，但这种作用的机制尚不清楚。一种可能性是Folates营救甲基化。尽管甲基化从未与神经rest发育有关，但我的初步数据表明，神经rest迁移需要甲基化。基于甲基转移酶EZH2在迁移神经rest细胞的细胞质中的表达，我假设非固定蛋白甲基化控制神经冠细胞的迁移特性。该建议旨在检验这一假设，并通过三个特定目的介导神经rest中甲基化的甲基化酶进行鉴定和表征：（1）三种候选甲基转移酶的空间和时间定位将通过immunofluorescence确定。 （2）反义吗啡寡核苷酸介导的蛋白消融将在神经rest发育过程中分析每个候选甲基转移酶的需求。 （3）使用显性负构建体评估甲基转移酶EZH2和细胞核中甲基转移酶EZH2的特定要求。一旦在神经波峰中确定了特定的甲基转移酶的需求，我们就可以开始研究叶酸缺陷对甲基化的影响，并阐明叶酸补充减少先天缺陷的机制。这项工作还将洞悉癌症转移的分子机制。公共卫生相关性：神经rest细胞是通过发育中的胚胎移动的细胞，形成神经系统，脸部，心脏和皮肤的一部分。这项研究的目的是确定神经rest细胞如何在整个体内移动。这个过程类似于癌症，将有助于我们了解癌症的进展，还将使我们更好地了解神经克雷斯特细胞运动中的缺陷如何导致出生缺陷。