NCI CLINICAL INVESTIGATOR AWARD PROGRAM

基因重排是体细胞可能是一个重要的 导致细胞恶性潜能的机制。 证据 这表明癌基因可能通过易位被激活， 多种人类恶性肿瘤与特定的 核型异常支持这一概念。 然而， 已知的机制，参与分子 重组或基因重组。 使用突变体 单纯疱疹病毒（HSV）作为探针，有可能开发一种 用于真核细胞中遗传重组的测定系统，和 将正常细胞与来自患有 DNA修复途径的异常 之间的重组 单纯疱疹病毒（HSV-1）突变体，其具有非重叠的 胸苷激酶或DNA中的互补突变 可以定量聚合酶基因座。 这些研究将决定 如果共感染HSV-1之间的遗传重组被调节 人类细胞的DNA损伤 抑制剂的作用 大分子合成（蛋白质合成和转录） 基因重组将有助于我们 了解这是否是一种诱导现象， 人体细胞 细胞功能的修饰剂如蛋白酶抑制剂、3- 氨基苯甲酰胺和佛波醇酯调节姐妹染色质 交换并可能影响DNA修复途径。 我们将 确定这些药物是否调节基因重组， 比较用以下物质预处理的细胞中的重组水平： 这两种药物中的任何一种都可以。 这些研究可能 有助于阐明那些细胞通路， 基因重组的表达。 在定量宿主细胞遗传重组的研究中， 具有特定DNA修复障碍的个体（例如，布鲁姆 综合征，范可尼贫血）将决定是否或 这两种突变体表达的重组水平不同， 在正常细胞中观察到的。 具体形式的影响 在基因重组时， 测定，沿着蛋白酶抑制剂和3- 在这种情况下的氨基苯甲酰胺。 为了确定这些细胞暴露是否先前被识别为 重组基因与突变子的表达有关 表型我们将采用诱变测定， HSV-1的ts突变体回复为ts+（非温度 敏感）表型。

Genetic rearrangement is somatic cells may be an important mechanism contributing to a cell's malignant potential. Evidence suggesting that oncogenes may be activated by translocation and the association of multiple human malignancies with a specific karyotypic abnormalities supports this concept. However, little is known about the mechanisms that are involved in molecular rearrangement or genetic recombination. Employing mutants of Herpes Simplex Virus (HSV) as probes it is possible to develop an assay system for genetic recombination in eukaryotic cells, and compare normal cells with those derived from patients having abnormalities in DNA repair pathways. Recombination between Herpes Simplex Virus (HSV-1) mutants having nonoverlapping complementary mutations in the thymidine kinase or DNA polymerase loci can be quantitated. These studies will determine if genetic recombination between co-infected HSV-1 is modulated by prior DNA damage to human cells. The effect of inhibitors of macromolecular synthesis (protein synthesis and transcription of mRNA) on genetic recombination will contribute to our understanding of whether this is an inducible phenomenon in human cells. Modifiers of cell function such as protease inhibitors, 3- aminobenzamide and phorbol esters modulate sister chromatic exchange and may affect DNA repair pathways. We will determine if these agents modulate genetic recombination by comparing the level or recombination in cells pretreated with either of these agents, with untreated cells. These studies may help elucidate those cellular pathways that are important for the expression of genetic recombination. In studies quantitating genetic recombination on host cells from individuals with specific DNA repair disorders (e.g., Bloom Syndrome, Fanconi's anemia) will determine whether either or both of these mutants express levels of recombination different from that observed in normal cells. The effects of specific forms of DNA damage to these cells on genetic recombination will be determined, along with the effects of protease inhibitors and 3- aminobenzamide in this setting. To determine if those cell exposures previously identified as recombinogenic are associated with expression of a mutator phenotype we will employ an assay for mutagenesis that measured reversions of ts mutants of HSV-1 to the ts+ (non-temperature sensitive) phenotype.