X chromosome cDNA microarray Screening and Functional Study of Novel XLMR genes

X染色体cDNA微阵列新型XLMR基因的筛选及功能研究

基本信息

批准号：
7494168
负责人：
TAO WANG
金额：
$ 23.91万
依托单位：
JOHNS HOPKINS 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：
2007-09-10 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7494168
关键词：
Accounting Affect Biological Assay Candidate Disease Gene Caring Cell Line Child Chromosomes, Human, X Clinical Clinical Management Cognition Collection Condition Counseling Data Defect Development Diagnosis Disabled Persons Disease Dissociation Economics Enzymes Exons Family Female Frameshift Mutation Gene Deletion General Population Genes Genetic Genetic Counseling Genetic Heterogeneity Goals Human Immunohistochemistry Impairment In Situ Hybridization In Vitro Individual Knowledge Link Luciferases Mediating Medical Mental Retardation Messenger RNA Molecular Mutation Nonsense Mutation Northern Blotting Nucleic Acid Regulatory Sequences Numbers Pathogenesis Patient Care Patients Persons Phenotype Physiological Pilot Projects Prevention Proteins RNA RNA Interference RNA Splicing Role Screening procedure Services Social Work Societies Supportive care Tertiary Protein Structure Transcript Transfection Variant Western Blotting X Chromosome X-Linked Mental Retardation base cDNA Arrays cognitive function cost design disability gene function genetic pedigree handicapping condition insight knock-down life time cost lymphoblast male novel novel strategies proband promoter segregation young adult

项目摘要

Mental retardation (MR) is the most common cause of handicaps in children and young adults and accounts for 2-3% in the general population. Patients with MR often require long-term medical and supportive care or services and accumulate enormous costs and burden to the families and the society. The average lifetime costs per person with MR were estimated to be more than $1 million. X-linked mental retardation (XLMR) occurs in 1 in 600 males and is genetically heterogeneous. Among the > 150-200 responsible loci on the X chromosome, <60 genes have been cloned. Identification of XLMR genes is essential for diagnosis, counseling, prevention, patient management, and rational development of effective therapy. It will also provide insight into the mechanism of human cognitive function. Our long-term goals are to understand the molecular basis and mechanism of X-linked mental retardation. In this application, we wish to capitalize on the exciting results from our pilot study of using a human X chromosome cDNA microarray (XCM) to identify novel XLMR genes. This XCM will be used to screen RNA from lymphoblasts of XLMR males to identify genes that show significant alternations in transcript levels. This strategy is based on the fact that a fraction of the mutations (estimated to be >30%) at any given locus result in dramatic alternations in the abundance of steady state transcripts. This approach is designed to detect mutations that result in a change in the abundance of mRNA due to mechanism such as promoter mutations, gene deletions or duplications, and abnormal RNA splicing associated with frameshift mutation and nonsense mutations associated with nonsense-mediated mRNA. It carries the advantage of fast screening of genes on the entire X chromosome without the need for large pedigree. Specifically we will (1) use XCM to screen 120 lymphoblast cell lines from XLMR males to identify novel candidate genes (2) identify responsible genetic defects by sequencing of the novel candidate genes in proband and in a large collection of XLMR males (3) functional studies of 4-6 novel XLMR genes to understand their physiological roles in human congnition and how mutations in these gene cause mental retardation. Results of the study will help to advance our knowledge on the genetic causes of mental retardation and to develop rational strategies for diagnosis, genetic counseling, prevention, and clinical care of patients with impairment of cognitive function..

精神发育迟滞 (MR) 是儿童和年轻人残疾的最常见原因。占总人口的 2-3%。 MR 患者通常需要长期的医疗和支持护理或服务，给家庭和社会积累了巨大的成本和负担。平均寿命每名 MR 患者的费用估计超过 100 万美元。 X 连锁智力低下 (XLMR) 男性发病率为六百分之一，并且具有遗传异质性。 X 上有 > 150-200 个负责位点染色体，已克隆<60个基因。 XLMR 基因的鉴定对于诊断至关重要，咨询、预防、患者管理以及合理制定有效的治疗方法。它还将提供对人类认知功能机制的深入了解。我们的长期目标是了解 X连锁智力低下的分子基础和机制。在此应用程序中，我们希望利用我们使用人类 X 染色体 cDNA 微阵列 (XCM) 来鉴定的初步研究结果令人兴奋新的 XLMR 基因。该 XCM 将用于筛选 XLMR 男性淋巴母细胞的 RNA，以鉴定转录水平显着改变的基因。该策略基于以下事实：任何给定位点的突变（估计>30%）都会导致丰度发生巨大变化稳态转录本。该方法旨在检测导致基因发生变化的突变。由于启动子突变、基因缺失或重复等机制导致的 mRNA 丰度，以及与移码突变相关的异常RNA剪接和与无义介导的 mRNA。具有快速筛选全X染色体基因的优势不需要庞大的血统。具体来说，我们将（1）使用XCM筛选120个淋巴母细胞系从 XLMR 男性中鉴定新的候选基因 (2) 通过对先证者和大量 XLMR 男性中的新候选基因 (3) 4-6 的功能研究新型 XLMR 基因，以了解它们在人类认知中的生理作用以及这些基因的突变如何基因导致智力低下。研究结果将有助于增进我们对遗传的认识精神发育迟滞的原因并制定合理的诊断、遗传咨询、预防策略以及认知功能障碍患者的临床护理。