DIRECT MUTAGENICITY TESTING IN MAN

人类直接突变性测试

• 批准号: 3169373
• 负责人: RICHARD J ALBERTINI
• 金额: $ 25.19万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 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/3169373
• 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

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.

本研究项目的目标是开发和验证 研究人体内体细胞突变的方法。 等 分析是必要的遗传风险估计，以及 回答有关分子本质的基本问题 人类的突变 关于突变机制的知识 对理解人类疾病有重要意义。 这些 研究旨在充分描述“自发”基因突变 发生在人体淋巴细胞中，并比较“自发” 在体内发生模型烷化剂诱导的基因突变 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-鸟嘌呤磷酸核糖基转移酶的研究 正常人外周血T淋巴细胞hprt基因突变 人，以及定义T细胞克隆的体内扩增 在正常个体中，被确定为“离群值”， 突变频率值（> 50 × 10-6）和HPRT的克隆性 变种人 (2)表征野生型和hprt突变T细胞 从正常个体的外周血回收的克隆 表面标志物表型，特异性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-苯丙氨酸氮芥诱导的hprt突变 在患有多发性骨髓瘤的患者的外周血淋巴细胞中， 骨髓瘤接受这种治疗。 (4)量化和 表征从以下获得的淋巴细胞中的体内hprt突变： 来自乳腺癌或骨髓瘤患者的引流淋巴结， 将结果与从外周血中获得的结果进行比较， 以比较分裂细胞和非分裂细胞中的突变。 (5)开发用于定量和表征的克隆测定 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基因座 突变频率的结果。 (6)开发一种用于 定量和表征体内产生的基因突变， 人B淋巴细胞，包括反映的克隆性定义 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基因座结果一致