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（申请人的描述）：该提案的主要目的是完成3个组成的职业开发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调查导致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员科学家的一年授权为他提供完成所需的指导和专业知识他的培训的学术和研究部分：2）一个教学计划专注于癌症生物学的特定领域； 3）过渡段从同事/导师到独立研究科学家和合作者在他的提议的独立研究部分。该提案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小儿恶性肿瘤的事件。假设申请人是，癌症儿童的体细胞突变事件将与较高的频率发生，并具有独特的突变光谱正常人口。将测试提议的具体目的假设是：1）确定背景和治疗诱导的儿童体内体细胞突变特定恶性肿瘤（急性淋巴细胞性白血病[ALL]，霍奇金氏病，神经母细胞瘤和肉瘤）并在可能的情况下将其关联随后疾病的分子事件：2）确定是否是否 “非法” V（D）J重组酶介导的突变发生在已知的癌症基因，p53; 3）隔离并识别T淋巴细胞模仿表型来自所有频率极高的儿童的克隆背景躯体突变。躯体突变事件的频率将由HPRT确定 T细胞克隆测定法。 HPRT和p53突变的突变光谱将是由多种方法确定，包括多重PCR，RT-PCR，IPCR， Southern印迹和DNA测序。超重克隆的克隆性将通过TCR伽马和CDR3区DNA序列的RFLP分析确定 TCR-β的分析。这些研究将提供有关频率和突变的第一个数据体内自发或治疗的频谱在患有特定恶性肿瘤的孩子。调查人员也将开始确定与背景相关的潜在诱变作用 “非法” V（d）J重组酶介导的事件，已知的癌症基因， p53。另外，突变表型克隆的隔离将允许表征遗传和生物学的未来研究与儿童白血病有关的缺陷。