SOMATIC MUTATIONS IN CHILDREN

儿童体细胞突变

• 批准号: 6343202
• 负责人: BARRY Alan FINETTE
• 金额: $ 10.97万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-08 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6343202
• 关键词: DNA damage; age difference; child (0-11); clinical research; environmental toxicology; gene frequency; gene mutation; genetic disorder; genetic mapping; human genetic material tag; human population genetics; human subject; human tissue; hypoxanthine phosphoribosyltransferase; molecular pathology; mutagens; newborn human (0-6 weeks); p53 gene /protein; passive smoking; tumor suppressor genes

DESCRIPTION: (Adapted from the Investigator's Abstract) The major objectives of this proposal are to study the mechanism and biological consequences of age-specific in vivo somatic genetic events in children. The hypothesis is that somatic mutations arising during fetal development and early childhood are more reflective of genetic changes with disease potential and environmental exposures than are mutations arising later in life. This hypothesis will be tested by three specific aims: 1) To complete studies comparing somatic mutational events that occur during fetal development and childhood with those that occur in adults; 2) To compare the prevalence of "illegitimate" V(D)J recombinase mediated mutations in a reporter gene, hprt, and a gene of disease significance, p53, and, 3) To determine the frequencies, spectra, and persistence of individual hprt mutations in newborns and children exposed to known genotoxic agents, and to correlate these molecular events with subsequent diseases. The frequency of somatic mutational events will be determined by the hprt T-cell cloning assay. Genetic analysis of hprt and p53 mutations will be performed using several methods: multiplex PCR, RT-PCR, IPCR, Southern blotting and DNA sequencing. These methods have previously been used for detailed molecular characterization of mutational events for inferences regarding genetic mechanisms and genotoxic monitoring. The research plan is designed to study the association/prediction between somatic mutations and specific diseases by determining: 1) whether the background frequency and mutational spectra of in vivo somatic mutations at the hprt locus in children are age-specific, reflecting unique biologic differences at the molecular level when compared to adults. These studies will provide information on the age dependent nature of mutational events in children and provide the database required for comparison studies in children with specific clinical diseases and genotoxic exposures; 2) if V(D)J recombinase mediated rearrangements captured at the hprt locus occur in a clinically specific tumor suppressor gene, p53. These studies may provide insights into the clinical effects of spontaneous age-specific mutagenic mechanism occurring during early human development; and 3) if somatic mutations induced by environmental exposures (cigarette smoke and chemotherapy) in children are predictive of subsequent somatic disease. The testing of potential mutagenic exposures in children is important since somatic mutational events during this period could have significant clinical consequences as well as long term effects as an adult. Results obtained from these studies will provide fundamental insights regarding the clinical relevance of age-specific, spontaneous and environmentally induced somatic mutations in children.

描述：(改编自《调查员摘要》)少校 本方案的目的是研究其发病机制和生物学特性。 儿童体内特定年龄的体细胞遗传事件的后果。 假设是在胎儿发育过程中产生的体细胞突变 早期儿童更能反映疾病的基因变化 潜在的和环境的暴露比后来出现的突变更多 生活。这一假设将通过三个具体目标进行检验：1) 比较胎儿期间发生的体细胞突变事件的完整研究 发展和童年与成人中发生的那些；2)比较 “不合法”V(D)J重组酶介导的突变在一种 报告基因hprt和与疾病相关的基因p53，以及，3)to 确定单个HPRT的频率、频谱和持久性 暴露于已知基因毒性物质的新生儿和儿童的突变，以及 将这些分子事件与随后的疾病联系起来。的频率 体细胞突变事件将由HPRT T细胞克隆决定 化验。HPRT和P53突变的基因分析将使用 几种方法：多重聚合酶链式反应、逆转录聚合酶链式反应、间接聚合酶链式反应、Southern杂交和DNA 测序。这些方法以前已经被用于详细的分子 突变事件的特征用于关于基因的推断 机制和遗传毒性监测。 研究计划旨在研究两者之间的关联/预测 通过确定：1)是否存在体细胞突变和特定疾病 体内体细胞突变的背景频率和突变谱 儿童的HPRT基因座具有年龄特异性，反映了独特的生物学特性 与成人相比，在分子水平上的差异。这些研究 将提供有关突变事件与年龄相关的性质的信息 并提供比较研究所需的数据库 有特定临床疾病和接触过基因毒性的儿童；2)如果 V(D)J重组酶介导的重排发生在HPRT基因座 在临床特定的肿瘤抑制基因P53中。这些研究可能 提供对特定年龄自发的临床效果的洞察 发生在人类早期发育过程中的诱变机制；3)如果 环境暴露诱发的体细胞突变(香烟烟雾和 在儿童中)可预示随后的躯体疾病。这个 对儿童进行潜在的诱变暴露测试很重要，因为 在此期间发生的体细胞突变事件可能具有重要的临床意义 作为成年人的后果和长期影响。取得的成果 这些研究将提供关于临床的基本见解 特定年龄、自发和环境诱导的体细胞的相关性 儿童的基因突变。