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IN VIVO SOMATIC MUTATIONS IN CHILDREN WITH CANCER

癌症儿童体内体细胞突变

基本信息

批准号：
6376745
负责人：
BARRY Alan FINETTE
金额：
$ 16.12万
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：
1998-09-30 至 2002-09-29
项目状态：
已结题

项目摘要

DESCRIPTION (Applicant's Description): The primary objective of this proposal is to complete a 3 component career development plan that will result in the applicant's acquiring the necessary skills and experience to develop an independent translational research program investigating the genetic mechanisms responsible for malignant transformation in children. The 3 components of this plan are: 1) a one-year co-mentorship with two senior cancer researcher scientists who will provide him with the guidance and expertise required to complete the academic and research portion of his training: 2) a didactic program that focuses on specific areas of cancer biology; and 3) a transition segment from colleague/mentor to independent research scientist and collaborator during the independent research portion of his proposal. The principal research objectives of this proposal are to investigate the molecular mechanisms and biologic consequences of in vivo somatic genetic events responsible for pediatric malignancies. The hypothesis of the applicants is that somatic mutational events in children with cancer will occur at a higher frequency and with a unique mutational spectra compared to a normal population. The specific aims which will test the proposed hypotheses are: 1) to determine the frequency and mutational spectra of background and t h erapy induced in vivo somatic mutations in children with specific malignancies (acute lymphocytic leukemia [ALL], Hodgkin's disease, neuroblastoma, and sarcomas) and where possible, to correlate these molecular events with subsequent diseases: 2) to determine if "illegitimate" V(D)J recombinase mediated mutations occur at a known cancer gene, p53; and 3) to isolate and identify T-lymphocyte imitator phenotype clones from children with relapsed ALL who have an extremely high frequency of background somatic mutations. The frequency of somatic mutational events will be determined by the hprt T-cell cloning assay. Mutational spectra of hprt and p53 mutations will be determined by a variety of methods including multiplex PCR, RT-PCR, IPCR, Southern blotting and DNA sequencing. Clonality of hypermutable clones will be determined by RFLP analysis of TCR gamma and CDR3 region DNA sequence analysis of TCR-beta. These studies will provide the first data on the frequency and mutational spectrum of in vivo spontaneous or treatment induced somatic mutations in children with specific malignancies. The investigators will also begin to determine the potential mutagenic effects associated [with] background "illegitimate" V(D)J recombinase mediated events in a known cancer gene, p53. In addition, the isolation of mutator phenotype clones will allow for future m e chanistic studies characterizing the genetic and biologic defect(s) associated with leukemogenesis in children.

描述（申请人描述）：本提案的主要目标是完成3个组成部分的职业生涯发展计划，将导致申请人获得必要的技能和经验，以发展独立的翻译研究研究恶性肿瘤的遗传机制的计划儿童的转变。该计划的三个组成部分是：1）a 与两名高级癌症研究科学家进行为期一年的共同指导，为他提供所需的指导和专业知识，他的训练的学术和研究部分：2）一个教学计划，专注于癌症生物学的特定领域;以及3）过渡部分从同事/导师到独立研究科学家和合作者在他的独立研究计划中本建议的主要研究目标是调查体内体细胞遗传的分子机制和生物学后果导致儿童恶性肿瘤的事件。的假设本申请人的一个观点是，癌症儿童的体细胞突变事件将与之相比，以更高的频率发生并具有独特的突变谱一个正常的人口。具体目标将测试拟议的假设是：1）确定频率和突变谱背景和激素诱导的儿童体内体细胞突变特异性恶性肿瘤（急性淋巴细胞白血病[ALL]，霍奇金病，神经母细胞瘤和肉瘤），并在可能的情况下，分子事件与随后的疾病：2）以确定是否 “非法”V（D）J重组酶介导的突变发生在已知的癌症中。 p53基因; 3）分离和鉴定T淋巴细胞模仿者表型来自复发ALL儿童的克隆，背景体细胞突变。体细胞突变事件的频率将由HPRT决定 T细胞克隆测定。将对hprt和p53突变的突变谱进行分析。通过多种方法包括多重PCR、RT-PCR、IPCR， Southern印迹和DNA测序。超变克隆的克隆性将通过TCR γ和CDR 3区DNA序列的RFLP分析确定 TCR-β的分析。这些研究将提供第一批关于频率和突变的数据。体内自发或治疗诱导的体细胞突变谱患有特定恶性肿瘤的儿童。调查人员还将开始确定与背景相关的潜在致突变效应已知癌症基因中的“非法”V（D）J重组酶介导的事件，第53页。此外，突变子表型克隆的分离将允许未来的遗传和生物学机制研究与儿童白血病发生相关的缺陷。