HEART AND LUNG DEFECTS IN DOWN SYNDROME

唐氏综合症的心脏和肺部缺陷

• 批准号: 2226112
• 负责人: JULIE RUTH KORENBERG
• 金额: $ 44.24万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2226112
• 关键词: Downs syndrome; artificial chromosomes; cardiopulmonary disease; chromosome 21; complementary DNA; congenital heart disorder; embryo /fetus cell /tissue; family genetics; gene expression; genetic library; genetic mapping; human fetus tissue; human genetic material tag; human tissue; in situ hybridization; laboratory mouse; lung disorder; medical complication; molecular cloning; nucleic acid sequence; pulsed field gel electrophoresis; regulatory gene; southern blotting; Downs syndrome; artificial chromosomes; cardiopulmonary disease; chromosome 21; complementary DNA; congenital heart disorder; embryo /fetus cell /tissue; family genetics; gene expression; genetic library; genetic mapping; human fetus tissue; human genetic material tag; human tissue; in situ hybridization; laboratory mouse; lung disorder; medical complication; molecular cloning; nucleic acid sequence; pulsed field gel electrophoresis; regulatory gene; southern blotting

The ultimate goal of the proposed research is the definition of genes responsible for congenital heart defects in Down syndrome. The proposed work represents a joint effort to narrow the region responsible for congenital heart defects in Down syndrome and then to isolate genes expressed from that region in an embryonic heart cDNA library. We have defined a small region (3-5 mb) of chromosome 21 likely to contain the gene(s) for DS-CHD. Based on a clue that both heart and lung defects in Down syndrome may result from the same tendency of Down syndrome fetal heart and lung fibroblasts to aggregate avidly, we will also prepare and screen cDNA libraries from trisomy 21 fetal heart and lung fibroblasts that show abnormal adhesion as well as from an embryonic lung cDNA library. The expression of candidate sequences by in situ hybridization in the developing endocardial cushions and/or pulmonary acinar cells will be utilized to determine candidate genes that could mediate the heart and lung defects in Down syndrome. If linked to chromosome 21, autosomal dominant pedigrees with endocardial cushion defects (ECD) will also be investigated and used to narrow the DS-CHD region. Thus, in this grant, we propose to: 1) Use cases involving partial aneusomy for chromosome 21 to further decrease the size of the DS-CHD region. In addition, we will utilize the trisomy 16 mouse that also develops endocardial cushion defects to delimit the size of the segment of chromosome 21 pathogenic for CHD. 2) Construct human embryonic heart and lung and trisomy 21 fetal cardiac and pulmonary fibroblast libraries as these represent the target tissues for cardiopulmonary defects in Down syndrome. 3) Screen these libraries and isolate genes by a novel recombination- based assay (RBA) we developed for this purpose. Portions of genes in this subregion will be identified by other methodologies such as computer analyses, exon amplification or hybrid selection and these sequences will be placed in our R6K supF plasmid, pMAD3. The RBA will then be performed both to confirm the tissue and time of expression and to isolate a longer gene fragment. 4) In situ hybridization of candidate genes will be performed to detect messages expressed in the endocardial cushions and/or acinar lung cells during organogenesis. Thus a gene "map" will be elaborated for sequences in a particular region of chromosome 21 known to be involved in abnormal cardiogenesis. At the least, this map will serve as a guideline for future studies to couple a "genic" initiative to characterize the time and tissue of transcription of particular regions to the "genomic" initiative. The efficient accomplishment of such a "genic" initiative will increase the value of the "genomic" initiative.

拟议研究的最终目标是基因的定义 负责唐氏综合症先天性心脏缺陷。 提议 工作代表了缩小负责地区的联合努力 唐氏综合症的先天性心脏缺陷，然后分离基因 在胚胎心脏cDNA文库中从该区域表达。 我们有 定义了一个小区域（3-5 MB）21的染色体可能包含 DS-CHD的基因。 基于心脏和肺部缺陷的线索 唐氏综合症的唐氏综合症胎儿的趋势可能导致唐氏综合症 心脏和肺成纤维细胞狂热地汇总，我们还将准备和 屏幕cDNA图书馆21胎儿心脏和肺成纤维细胞 它显示异常粘附以及胚胎肺cDNA 图书馆。 通过原位杂交表达候选序列 在发展中的心内膜垫和/或肺腺泡细胞中 用于确定可以介导心脏和的候选基因 唐氏综合症的肺部缺陷。 如果链接到21号染色体，常染色体 具有心内垫缺陷（ECD）的主要血统也将是 研究并用于缩小DS-CHD区域。 因此，在这笔赠款中，我们建议： 1）涉及21染色体的部分动脉瘤的用例以进一步 减小DS-CHD区域的大小。 此外，我们将 利用也会发展出心内膜垫的三体鼠标 缺陷以划定染色体21的片段的大小 冠心病的致病性。 2）构建人类胚胎心脏，肺和三体胎21胎儿 心脏和肺成纤维细胞库，因为它们代表 针对唐氏综合症的心肺缺陷的靶向组织。 3）通过新的重组筛选这些文库和分离基因 我们为此目的开发的基于测定（RBA）。 基因的部分 在此子区域中，将由其他方法（例如 计算机分析，外显子扩增或混合选择，这些 序列将放置在我们的R6K SUPF质粒PMAD3中。 RBA 然后将执行以确认组织和时间 表达并分离出更长的基因片段。 4）将进行候选基因的原位杂交 检测内膜垫和/或腺泡中表达的消息 器官发生过程中的肺部细胞。 因此，将在特定区域详细阐述基因“映射” 已知参与异常心脏发生的21染色体。 在 至少，这张地图将作为未来研究的指南 特征转录的时间和组织的“基因”计划 “基因组”倡议的特定区域。 高效 完成这样的“基因”计划将增加 “基因组”倡议。