Genomic studies of CHD7 in CHARGE syndrome

CHARGE 综合征中 CHD7 的基因组研究

基本信息

批准号：
8100186
负责人：
Peter Christopher Scacheri
金额：
$ 30.58万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-20 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8100186
关键词：
11p15 Affect Binding Binding Proteins Binding Sites Biological Assay Body Patterning CHARGE syndrome Cell Culture Techniques Cells Cessation of life Chromatin Chromosomes Clinical Code Coloboma Congenital Abnormality Congenital Heart Defects Cranial Nerves Data Development Disease Ear Embryonic Development Epigenetic Process Equilibrium Ethylnitrosourea Etiology Functional RNA Gene Expression Gene Targeting Genes Genetic Transcription Genomics Growth H19 gene HOXA10 gene Histone H3 Histones Homeobox Genes Human Human Development In Vitro Infant Insulin-Like Growth Factor II Investigation Knowledge Lead Learning Link Location Lysine Maps Mediating Modeling Molecular Molecular Profiling Mus Mutagenesis Mutant Strains Mice Mutation Neural Pathways Nuclear Protein Organ Pathogenesis Pathway interactions Patients Proteins RNA Interference Recombinants Recruitment Activity Research Research Personnel Role Screening procedure Site Specificity Staging Syndrome Techniques Testing Time Transcriptional Regulation Vision body system chromatin immunoprecipitation chromatin modification design embryonic stem cell feeding fetal hearing impairment helicase histone modification human disease imprint in vivo loss of function loss of function mutation malformation mouse model mutant orofacial postnatal premature programs promoter

项目摘要

DESCRIPTION (provided by applicant): CHARGE syndrome is a congenital disease characterized by malformations of multiple organs. ~70% of CHARGE syndrome cases are caused by loss-of-function de novo mutations in the CHD7 gene (coding for chromodomain helicase DMA-binding protein 7). Little information is available about the normal function of the CHD7 protein and its role in human development and disease. Our preliminary studies demonstrate that CHD7 is a nuclear protein that directly binds to multiple genes, including HOX genes (HOXA5, HOXA10, and HOXA11) and imprinted genes (IGF2 and H19) that are essential for normal embryonic development. The proposed research tests the hypothesis that the malformations seen in patients with CHARGE syndrome are caused by aberrant transcription of specific CHD7 target genes. This hypothesis will be tested in 3 Specific Aims. In Aim 1, we will evaluate a subset of the CHD7 target genes to determine if CHD7 directly regulates their expression. Specifically, expression of 50 CHD7 targets will quantified in cell culture before and after knockdown of CHD7 by RNAi. In addition, to determine if anomalies in CHARGE syndrome are due to dysregulated expression of HOX, Igf2, and H19, we will analyze expression of these genes in developing Chd7 mutant mice that are an excellent model CHARGE syndrome. In Aim 2, we will investigate the mechanism by which CHD7 is recruited to its target genes, using in vitro and in vivo assays designed to reveal interactions between CHD7 and various histone modifications on chromatin. In Aim 3, we will identify CHD7 targets that directly depend on CHD7 during early development, using an unbiased genomics approach that combines the technique of chromatin immunoprecipitation on microarrays (ChlP-chip) with expression profiling of wild type and mutant mouse ES cells. By identifying and characterizing the genes that are directly regulated by CHD7, we expect to learn more about (1) normal human development, (2) the causes of the isolated birth defects that make up the spectrum of anomalies in CHARGE, and (3) the etiology of this rare syndrome. In addition to furthering our understanding of the clinical implications of genes regulated by CHD7, we anticipate gaining a significant amount of knowledge about the molecular mechanisms of transcriptional regulation by delineating the interactions between CHD7 and its target genes.

描述（由申请人提供）： CHARGE综合征是一种以多器官畸形为特征的先天性疾病。大约 70% 的 CHARGE 综合征病例是由 CHD7 基因（编码染色质解旋酶 DMA 结合蛋白 7）的功能缺失突变引起的。关于 CHD7 蛋白的正常功能及其在人类发育和疾病中的作用的信息很少。我们的初步研究表明，CHD7 是一种核蛋白，可直接与多个基因结合，包括正常胚胎发育必需的 HOX 基因（HOXA5、HOXA10 和 HOXA11）和印记基因（IGF2 和 H19）。拟议的研究测试了以下假设：CHARGE 综合征患者的畸形是由特定 CHD7 靶基因的异常转录引起的。这一假设将在 3 个具体目标中得到检验。在目标 1 中，我们将评估 CHD7 靶基因的子集，以确定 CHD7 是否直接调节其表达。具体而言，将在通过 RNAi 敲除 CHD7 之前和之后的细胞培养物中对 50 个 CHD7 靶标的表达进行定量。此外，为了确定 CHARGE 综合征的异常是否是由于 HOX、Igf2 和 H19 表达失调所致，我们将分析这些基因在发育中的 Chd7 突变小鼠中的表达，这些小鼠是 CHARGE 综合征的优秀模型。在目标 2 中，我们将使用旨在揭示 CHD7 与染色质上各种组蛋白修饰之间相互作用的体外和体内测定来研究 CHD7 被招募到其靶基因的机制。在目标 3 中，我们将使用无偏基因组学方法，将微阵列染色质免疫沉淀技术（ChlP 芯片）与野生型和突变型小鼠 ES 细胞的表达谱相结合，确定早期发育过程中直接依赖于 CHD7 的 CHD7 靶点。通过识别和表征受 CHD7 直接调节的基因，我们期望了解更多关于 (1) 正常人类发育，(2) 构成 CHARGE 异常谱的孤立出生缺陷的原因，以及 (3)这种罕见综合征的病因。除了进一步了解 CHD7 调控基因的临床意义外，我们预计通过描述 CHD7 与其靶基因之间的相互作用，可以获得有关转录调控分子机制的大量知识。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mutations in the CHD7 gene: the experience of a commercial laboratory.

CHD7 基因突变：商业实验室的经验。

DOI：
10.1089/gtmb.2010.0101
发表时间：
2010-12-15
期刊：
Genetic testing and molecular biomarkers
影响因子：
0
作者：
C. Bartels;C. Scacheri;L. White;P. Scacheri;S. Bale
通讯作者：
S. Bale

ITCH K63-ubiquitinates the NOD2 binding protein, RIP2, to influence inflammatory signaling pathways.

ITCH K63 泛素化 NOD2 结合蛋白 RIP2，以影响炎症信号传导通路。

DOI：
发表时间：
2009-08-11
期刊：
影响因子：
0
作者：
Tao, Mingfang;Scacheri, Peter C;Marinis, Jill M;Harhaj, Edward W;Matesic, Lydia E;Abbott, Derek W
通讯作者：
Abbott, Derek W

Epigenetic signatures distinguish multiple classes of enhancers with distinct cellular functions.

表观遗传特征区分了具有不同细胞功能的多类增强子。