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：
7305496
负责人：
TAO WANG
金额：
$ 24.4万
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/7305496
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

DESCRIPTION (provided by applicant): 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 alterations 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 cognition 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.

描述（由申请人提供）：精神发育迟滞 (MR) 是儿童和青少年残疾的最常见原因，占总人口的 2-3%。 MR患者往往需要长期的医疗和支持性护理或服务，给家庭和社会带来巨大的成本和负担。据估计，每名 MR 患者的平均终生费用将超过 100 万美元。 X 连锁智力低下 (XLMR) 的发生率是每 600 名男性中就有 1 人发生，并且具有遗传异质性。 X染色体上>150-200个负责位点中，<60个基因已被克隆。 XLMR 基因的鉴定对于诊断、咨询、预防、患者管理和合理开发有效疗法至关重要。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) 来鉴定新型 XLMR 基因的初步研究的令人兴奋的结果。该 XCM 将用于筛选 XLMR 雄性淋巴母细胞的 RNA，以识别转录水平发生显着变化的基因。该策略基于以下事实：任何给定位点的一小部分突变（估计>30%）会导致稳态转录本丰度发生巨大变化。该方法旨在检测由于启动子突变、基因缺失或重复等机制导致 mRNA 丰度变化的突变，以及与移码突变相关的异常 RNA 剪接和与无义介导的 mRNA 相关的无义突变。它的优点是可以快速筛选整个X染色体上的基因，而不需要庞大的谱系。具体来说，我们将 (1) 使用 XCM 筛选来自 XLMR 男性的 120 个淋巴母细胞系，以识别新的候选基因 (2) 通过对先证者和大量 XLMR 男性中的新候选基因进行测序来识别负责任的遗传缺陷 (3) 功能性基因缺陷研究 4-6 个新型 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导致智力低下。该研究结果将有助于增进我们对智力低下遗传原因的认识，并为认知功能障碍患者的诊断、遗传咨询、预防和临床护理制定合理的策略。