SOMATIC MUTATIONS IN CHILDREN

儿童体细胞突变

基本信息

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
项目类别：
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

项目摘要

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其机制和生物学儿童体内特定年龄体细胞遗传事件的后果。该假设是胎儿发育过程中发生的体细胞突变和幼儿期更能反映疾病引起的基因变化潜在和环境暴露比稍后发生的突变生活。该假设将通过三个具体目标进行检验：1）比较胎儿时期发生的体细胞突变事件的完整研究发育和童年与成人发生的情况相同； 2）比较 “非法”V(D)J 重组酶介导突变的普遍存在报告基因 hprt 和具有疾病意义的基因 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通过 hprt T 细胞克隆来确定化验。 hprt 和 p53 突变的遗传分析将使用多种方法：多重 PCR、RT-PCR、IPCR、Southern 印迹和 DNA 测序。这些方法以前已用于详细的分子突变事件的表征，用于遗传推论机制和遗传毒性监测。该研究计划旨在研究之间的关联/预测体细胞突变和特定疾病通过确定：1）是否体内体细胞突变的背景频率和突变谱儿童的 hprt 位点具有年龄特异性，反映了独特的生物学特性与成人相比，分子水平上的差异。这些研究将提供有关突变事件年龄依赖性的信息儿童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 位点捕获的 V(D)J 重组酶介导的重排发生临床特异性肿瘤抑制基因 p53。这些研究可能提供对自发年龄特定的临床效果的见解人类早期发育过程中发生的诱变机制； 3）如果环境暴露（香烟烟雾和儿童的化疗）可预测随后的躯体疾病。这测试儿童潜在的致突变暴露非常重要，因为这一时期的体细胞突变事件可能具有重大的临床意义后果以及作为成年人的长期影响。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相关性儿童的突变。