SOMATIC MUTATIONS IN CHILDREN

儿童体细胞突变

• 批准号: 2472538
• 负责人: BARRY Alan FINETTE
• 金额: $ 9.84万
• 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/2472538
• 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); 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：（改编自研究者摘要）主要 该提案的目的是研究机制和生物学 儿童体内年龄特异性体细胞遗传事件的后果。 假设是胎儿发育过程中产生的体细胞突变 和幼儿期更能反映疾病的遗传变化 潜在的和环境的暴露比突变产生后， 生活 这一假设将通过三个具体目标进行检验：1） 完整的研究比较了胎儿期发生的体细胞突变事件， 2）将儿童期和儿童期的发育与成人期的发育进行比较; “非法”V（D）J重组酶介导的突变在一个 报告基因hprt和疾病显著性基因p53，以及，3）为了 确定单个HPRT的频率、频谱和持续时间 暴露于已知遗传毒性物质的新生儿和儿童中的突变，以及 将这些分子事件与随后的疾病联系起来。 的频率 体细胞突变事件将通过hprt T细胞克隆来确定 比色法 将使用以下方法进行hprt和p53突变的遗传分析： 多种方法：多重PCR、RT-PCR、IPCR、Southern blotting和DNA 测序 这些方法以前曾用于详细的分子 突变事件的表征，用于关于遗传学的推断 机制和遗传毒性监测。 研究计划旨在研究以下因素之间的关联/预测： 体细胞突变和特定疾病，通过确定：1）是否 背景频率和突变谱的体内体细胞突变， 儿童HPRT基因座是年龄特异性的，反映了独特的生物学特性 在分子水平上与成年人相比存在差异。 这些研究 将提供关于突变事件的年龄依赖性的信息， 并提供比较研究所需的数据库， 患有特定临床疾病和遗传毒性暴露的儿童; 2）如果 在hprt基因座捕获的V（D）J重组酶介导的重排发生 一种临床特异性肿瘤抑制基因p53 这些研究可能 提供了对自发性年龄特异性 在人类早期发育过程中发生的致突变机制;以及3）如果 环境暴露（香烟烟雾和 化疗）是预测随后的躯体疾病。 的 测试儿童的潜在诱变暴露是重要的，因为 在此期间发生的体细胞突变事件可能具有显著的临床意义。 作为一个成年人的长期影响。 获得的结果 这些研究将提供有关临床的基本见解， 年龄特异性、自发性和环境诱导的躯体 孩子们的突变。