Gene Mutation and Rescue in Human Diaphragmatic Hernia

人类膈疝的基因突变与挽救

• 批准号: 7892730
• 负责人: PATRICIA K DONAHOE
• 金额: $ 1.2万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7892730
• 关键词: Affect; Age; Animal Model; Animals; Bacterial Artificial Chromosomes; Biochemical; Biochemical Pathway; Biological Assay; Birth; Blood; Boston; Candidate Disease Gene; Cell Line; Cells; Cheek structure; Clinical; Code; Complex; Congenital diaphragmatic hernia; Cytogenetics; DNA; DNA Sequence Rearrangement; Databases; Defect; Development; Diagnosis; Diaphragmatic Hernia; Drosophila genus; Equilibrium; Family; Family member; Fetal Lung; Gene Mutation; Generations; Genes; Genetic; Genome; Genomics; Genotype; Hot Spot; Human; Immunohistochemistry; In Situ Hybridization; Intervention; Knowledge; Ligands; Loss of Heterozygosity; Lung; Metaphase; Molecular; Molecular Genetics; Monozygotic Twinning; Monozygotic twins; Mus; Mutate; Mutation; Organ Culture Techniques; Parents; Pathology; Patient Care; Patients; Pediatric Hospitals; Phenotype; Preparation; Proteins; Recruitment Activity; Research Personnel; Resolution; Respiratory Diaphragm; Role; Siblings; Site; Source; Specimen; Structure of parenchyma of lung; Surgeon; Swab; Syndrome; Techniques; Testing; Therapeutic; Therapeutic Intervention; Tissue Sample; Unbalanced Translocation; base; cohort; comparative genomic hybridization; design; established cell line; experience; genetic analysis; genetic linkage analysis; genome-wide; improved; indexing; kindred; malformation; member; microdeletion; mouse model; patient population; prevent; programs; tissue culture; tool

Congenital Diaphragmatic Hernia (CDH) is a frequent and often fatal developmental condition of diaphragm defects associated with lung hypoplasia caused by diverse factors that we hypothesize are predominantly genetic, but heterogeneous. Project IV will use a combinationof clinical, molecular, developmental, genomic and cytogenetic strategies to identify mutations causing CDH, with the hope of elucidating perturbed biochemical pathways that can then serve as functional targets for pharmacological intervention or treatment. Aim I: We will recruit a cohort of carefully phenotyped isolated and complex CDH patients from two major cljnica! sites, MGHand Children's Hospital Boston, establish cell lines on each patient, and parents, and siblings, and enter deceased populations of patients, families with multiple affected members, consanguineous families, and monozygotic twins. DMA will be extracted from various sources for mutational analysis of candidate genes, arrayed based Comparative Genomic Hybridization (aCGH), loss of heterozygosity studies (LOH), and metaphase preparations for subtelomeric FISH. Aim II: High resolution cytogenetic tools such as 1 Mbarray CDH, subtelomeric FISH, and multiplex igand-directed probe amplification (MLPA) will be used to identify microdeletions, microduplications. and balanced and unbalanced translocations followed by breakpoint analyses to identify genes in these regions hat have mutations causative for CDH. Aim III: Nonsynonomous SNPs identified will be studied as plausible causative mutations. Genome wide SNP analysis will be used to reveal regions of LOH in consanguineous Donnai Barrow kindreds and in discordant monozygotic twins. Aim IV: Candidate genes from animal models or from LOH regions with CDH in human will be studied br abnormal expression in human CDH lungs or diaphragms and tested in Drosophila or mouse cell-based or organ culture assays to determine functional significance. CDH gene defects uncovered by these studies will serve as targets for pharmacological or other therapeutic interventionsto ameliorate or prevent this severe birth malformation. - .

先天性膈疝（CDH）是一种常见的，往往是致命的膈肌发育条件 与肺发育不全相关的缺陷，由多种因素引起，我们假设主要是 遗传的，但异质的Project IV will use a combination组合of clinical临床，molecular分子，developmental发展，genomic基因 和细胞遗传学策略，以确定导致CDH的突变，希望阐明干扰 然后可以作为药理学干预的功能靶标的生化途径，或 治疗 目的一：我们将从两个国家招募一组经过仔细分型的孤立和复杂CDH患者， cljnica少校!地点，波士顿MGHand儿童医院，在每个病人和父母身上建立细胞系， 和兄弟姐妹，并进入死亡的患者群体，有多个受影响成员的家庭， 近亲家庭和同卵双胞胎将从各种来源提取DMA进行突变 候选基因分析，基于阵列的比较基因组杂交（aCGH）， 杂合性研究（洛）和亚端粒FISH的中期制备。 目的II：高分辨率的细胞遗传学工具，如1 Mbarray CDH，亚端粒FISH和多重 配体定向探针扩增（MLPA）将用于鉴定微缺失、微重复。和 平衡和不平衡易位，随后进行断点分析以鉴定这些区域中的基因 都有导致先天性髋关节脱位的突变 目的III：将识别的非同义SNP作为可能的致病突变进行研究。全基因组 SNP分析将用于揭示近亲Donnai巴罗家系和近亲Donnai家系中的洛缺失区域。 不协调的单卵双胞胎 目的四：研究CDH动物模型或人类CDH洛区的候选基因 br在人CDH肺或横膈膜中的异常表达，并在果蝇或小鼠细胞中进行了检测 或器官培养测定以确定功能意义。这些研究发现的CDH基因缺陷 将作为药理学或其他治疗干预的靶点来改善或预防这种情况 严重的出生畸形- .