Characterization of Enhancers in the Epidermal Differentiation Complex

表皮分化复合物中增强剂的表征

• 批准号: 8307242
• 负责人: Cristina de Guzman Strong
• 金额: $ 23.7万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-16 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8307242
• 关键词: 1q21; Affect; American; Atopic Dermatitis; Binding; Bioinformatics; Biological Assay; Canis familiaris; Cells; Chromatin; Chromosomes; Complex; Development Plans; Didelphidae; Disease; EP300 gene; Embryo; Enhancers; Epidermis; Epigenetic Process; Gene Cluster; Gene Duplication; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genome; Globin; Goals; Human; Human Chromosomes; Hypersensitivity; In Vitro; Inflammatory; Institution; Laboratories; LacZ Genes; Light; Link; Locus Control Region; Luciferases; Macaca mulatta; Maps; Mentors; Mentorship; Mus; National Human Genome Research Institute; Pan Genus; Phase; Positioning Attribute; Process; Psoriasis; Quality of life; Rattus; Regulation; Regulatory Element; Reporter; Research; Sequence Alignment; Sequence Analysis; Skin; Staging; Stress; Testing; Transcriptional Regulation; Transgenic Mice; Universities; Washington; base; career development; chromatin immunoprecipitation; comparative; cost; density; genome sequencing; in vivo; keratinocyte; mouse model; promoter; skin disorder; transcription factor

Atopic dermatitis (AD) and psoriasis are two inflammatory skin diseases that affect over 22.5 million Americans collectively and are burdening the U.S. economy with over $7.5 billion dollars in management and quality-of-life issue costs. These diseases share linkage to the epidermal differentiation complex (EDC) on chromosome 1q21. The EDC spans 1.6Mb and contains a majority of genes that are specifically expressed upon terminal differentiation of the epidermis and subsequent skin barrier formation. The proximity and density of these genes on 1q21 suggest coordinate regulation via cis-regulatory elements. To date, only a few regulatory elements have been identified in the EDC. My goal is to identify enhancers that control gene expression in the EDC. Using comparative multi-species sequence analysis, we hypothesize that evolutionarily conserved noncoding sequences (CNS) function as enhancers to coordinate transcriptional regulation of genes within the EDC. To test this hypothesis, we propose the following 2 aims: In Specific Aim 1, we will test the function of the CNS to direct in vitro enhancer activity using luciferase reporter assays in mouse epidermal cells. In Specific Aim 2A, we will determine colocalization of the CNS EDC to open chromatin using DNasel hypersensitivity assays in mouse keratinocytes and barrier deficient mouse models. These aims will be completed during the mentored phase under Dr. Julie Segre at the National Human Genome Research Institute (NHGRI) in Bethesda, MD. The following aims will be pursued during the independent phase under the bridge mentorship of Dr. Anne Bowcock from Washington University: In Specific Aim 2B, we will also determine if these CNS EDC map to regions bound by p300 and monomethylated H3K4 enriched in enhancer sequences using chromatin immunoprecipitation (ChIP). In Specific Aim 3, will assess transcription factor binding to the CNS EDC using bioinformatics and ChIP. In Specific Aim 4, we will test select CNS EDC in vivo as epidermal locus control regions using CNS EDC- hsp68-lacZ transgenic mice. This proposal functions as Cristina de Guzman Strong's career development plan in obtaining an independent research position at an academic institution. Results from the research plan will shed light on the genetic and epigenetic mechanisms underlying epidermal differentiation and barrier formation as well contribute to our understanding of atopic dermatitis and psoriasis.

特应性皮炎（AD）和银屑病是两种炎症性皮肤病，影响超过2250万人 美国人共同承担着超过75亿美元的管理费用， 生活质量问题的成本。这些疾病与表皮分化复合体（EDC）有关， 染色体1q21。EDC跨越1.6Mb，包含大部分特异性表达的基因， 在表皮的终末分化和随后的皮肤屏障形成时。接近和 这些基因在1q21上的密度表明通过顺式调节元件进行协调调节。到目前为止，只有一个 在EDC中确定了一些监管要素。我的目标是找出控制基因的增强子 在EDC中表达。利用比较多物种序列分析，我们假设， 进化上保守的非编码序列（CNS）作为增强子协调转录 EDC内的基因调控。为了验证这一假设，我们提出了以下两个目标： 1中，我们将使用荧光素酶报告基因测定来测试CNS指导体外增强子活性的功能， 小鼠表皮细胞在特定目标2A中，我们将确定CNS EDC的共定位，以打开 在小鼠角质形成细胞和屏障缺陷小鼠模型中使用DNase1超敏性测定染色质。 这些目标将在国家人类学中心的Julie塞格雷博士的指导下完成。 位于马里兰州贝塞斯达的基因组研究所（NHGRI）。会议期间将努力实现以下目标： 在华盛顿大学Anne Bowcock博士的桥梁指导下， 具体目标2B，我们还将确定这些CNS EDC是否映射到p300结合的区域， 使用染色质免疫沉淀法（ChIP）测定富集增强子序列的单甲基化H3K4。在 具体目标3将使用生物信息学和ChIP评估转录因子与CNS EDC的结合。在 具体目标4，我们将使用CNS EDC在体内测试选择的CNS EDC作为表皮基因座控制区。 hsp68-lacZ转基因小鼠。这份提案的功能是Cristina de Guzman Strong的职业发展 在学术机构获得独立研究职位的计划。研究结果 该计划将阐明表皮分化的遗传和表观遗传机制， 屏障的形成也有助于我们理解特应性皮炎和银屑病。

项目成果

The Molecular Revolution in Cutaneous Biology: EDC and Locus Control.

皮肤生物学的分子革命：EDC 和基因座控制。

• DOI: 10.1016/j.jid.2016.03.046
• 发表时间: 2017
• 期刊: The Journal of investigative dermatology
• 作者: Oh,InezY;deGuzmanStrong,Cristina
• 通讯作者: deGuzmanStrong,Cristina