DIRECT MUTAGENICITY TESTING IN MAN

人类直接突变性测试

基本信息

批准号：
3169368
负责人：
RICHARD J ALBERTINI
金额：
$ 25.88万
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：
1981
资助国家：
美国
起止时间：
1981-09-01 至 1993-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3169368
关键词：
6 thioguanine B cell receptor Epstein Barr virus T lymphocyte alkylating agents autosomal recessive trait biopsy blood /lymphatic neoplasm cancer risk carcinogen testing cell transformation clone cells combination cancer therapy congenital disorders cytogenetics drug related neoplasm /cancer drug resistance early diagnosis environment related neoplasm /cancer enzyme linked immunosorbent assay gene frequency gene mutation gene rearrangement genetic markers histocompatibility typing human population genetics human subject hypoxanthine phosphoribosyltransferase immunogenetics immunoglobulin genes immunosuppression interleukin 2 interleukin 4 lymphocyte major histocompatibility complex messenger RNA molecular cloning multiple myeloma mutagen testing mutagens mutant natural gene amplification neoplasm /cancer diagnosis neoplasm /cancer genetics phenylalanine point mutation receptor site directed mutagenesis ultraviolet radiation viral carcinogenesis

项目摘要

The objective of this research program is to develop and validate methods for studying in vivo somatic cell mutation in humans. Such assays are necessary for genetic risk estimates, as well as answering basic questions concerning the molecular nature of mutation in humans. Knowledge of the mechanism of mutation has important implications for understanding human disease. These studies aim to fully characterize "spontaneous" gene mutation occurring in vivo in human lymphocytes, and compare "spontaneous" with model alkylating agent induced gene mutation occurring in vivo in human lymphocytes. These objectives will be attained by achieving the following specific aim: (1) Completing population studies of in vivo hypoxanthine-guanine phosphoribosyltransferase gene (hprt) mutation in peripheral blood T-lymphocytes in normal humans, as well as defining in vivo amplifications of T-cell clones in normal individuals, ascertained as "outliers" with respect to mutant frequency values (> 50 x 10-6) and clonality of hprt mutants. (2) Characterizing wild type and hprt mutant T-cell clones recovered from the peripheral blood of normal individuals of surface marker phenotypes, specific T-cell receptor (TRC) gene rearrangement patterns, and hprt gene alterations. (3) Quantifying and characterizing L-phenylalanine mustard induced hprt mutation in vivo in peripheral blood lymphocytes of patients with multiple myeloma receiving such treatment. (4) Quantifying and characterizing in vivo hprt mutation in lymphocytes obtained from draining lymph nodes from breast cancer or myeloma patients and comparing findings with those obtained from peripheral blood in order to compare mutation in dividing versus non-dividing cells. (5) Developing a clonal assay for quantifying and characterizing gene mutations arising in vivo at the autosomal HLA locus in human lymphocytes, for comparison and with the X-linked hprt locus mutation frequency results. (6) Developing a clonal assay for quantifying and characterizing gene mutations arising in vivo in human B-lymphocytes, including definition of clonality as reflected by immunoglobulin (Ig) gene rearrangement patterns, for comparison with the T-lymphocyte hprt locus results.

该研究计划的目标是开发和验证研究人类体内体细胞突变的方法。这样的分析对于遗传风险评估是必要的，以及回答有关分子性质的基本问题人类的突变。对突变机制的了解对于理解人类疾病具有重要意义。这些研究旨在全面表征“自发”基因突变发生在人体淋巴细胞体内，并比较“自发” 模型烷化剂诱导体内发生基因突变在人类淋巴细胞中。这些目标将通过以下方式实现实现以下具体目标： (1) 完成人口体内次黄嘌呤鸟嘌呤磷酸核糖转移酶的研究正常人外周血T淋巴细胞基因（hprt）突变人类，以及定义 T 细胞克隆的体内扩增在正常个体中，被确定为“异常值” 突变频率值 (> 50 x 10-6) 和 hprt 克隆性突变体。 (2) 表征野生型和hprt突变T细胞从正常人的外周血中回收的克隆表面标记表型、特定 T 细胞受体 (TRC) 基因重排模式和 hprt 基因改变。 (3)量化并表征 L-苯丙氨酸芥子诱导的 hprt 突变体内多发性硬化症患者的外周血淋巴细胞骨髓瘤接受这样的治疗。 (4) 量化和表征从获得的淋巴细胞中获得的体内 HPRT 突变引流乳腺癌或骨髓瘤患者的淋巴结，以及与从外周血获得的结果进行比较为了比较分裂细胞与非分裂细胞的突变。 (5) 开发用于定量和表征的克隆测定法人类常染色体 HLA 基因座体内发生的基因突变淋巴细胞，用于比较并与 X 连锁 hprt 位点突变频率结果。 (6) 开发克隆分析量化和表征体内产生的基因突变人类 B 淋巴细胞，包括所反映的克隆性定义通过免疫球蛋白 (Ig) 基因重排模式进行比较与T淋巴细胞hprt位点结果。