Genomic studies of CHD7 in CHARGE syndrome

CHARGE 综合征中 CHD7 的基因组研究

• 批准号: 8099335
• 负责人: Peter Christopher Scacheri
• 金额: $ 10.21万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-10 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8099335
• 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; Etiology; Functional RNA; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Genomics; Growth; H19 gene; HOXA10 gene; 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; Nuclear Proteins; 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; imprint; in vivo; loss of function; loss of function mutation; malformation; mouse model; mutant; orofacial; postnatal; premature; programs; promoter

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 miroarrays (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.

CHARGE综合征是一种以多器官畸形为特征的先天性疾病。~70%的 CHARGE综合征病例是由CHD 7基因（编码 染色体结构域解旋酶DMA结合蛋白7）。关于正常功能的信息很少 CHD 7蛋白及其在人类发育和疾病中作用。我们的初步研究表明， CHD 7是直接结合多个基因的核蛋白，所述多个基因包括HOX基因（HOXA 5、HOXA 10和HOXA 11）。 HOXA 11）和印记基因（IGF 2和H19）是正常胚胎发育所必需的。的 拟议的研究测试了这样的假设：CHARGE综合症患者中观察到的畸形是 由特异性CHD 7靶基因的异常转录引起。该假设将在3个特定的 目标。在目标1中，我们将评估CHD 7靶基因的一个子集，以确定CHD 7是否直接调节 他们的表情。具体地，将在细胞培养之前和之后定量50种CHD 7靶标的表达。 通过RNAi敲低CHD 7。此外，为了确定CHARGE综合征中的异常是否是由于 HOX，Igf 2和H19的表达失调，我们将分析这些基因在发育中的表达。 Chd 7突变小鼠是一种极好的CHARGE综合征模型。在目标2中，我们将研究 CHD 7被募集到其靶基因的机制，使用体外和体内试验， 揭示了CHD 7与染色质上各种组蛋白修饰之间的相互作用。在目标3中，我们将确定 CHD 7靶点在早期发育过程中直接依赖于CHD 7，使用无偏倚的基因组学 将微阵列上的染色质免疫沉淀技术（ChIP芯片）与 野生型和突变型小鼠ES细胞的表达谱。通过识别和描述基因 直接受CHD 7调节，我们希望更多地了解（1）正常的人类发育，（2） 构成CHARGE异常谱的孤立出生缺陷的原因，以及（3） 这种罕见的综合征的病因。除了加深我们对临床意义的理解， 通过研究CHD 7调控的基因，我们预计将获得大量关于CHD 7分子调控的知识。 通过描绘CHD 7与其靶基因之间的相互作用来研究转录调控机制。