BIOMARKERS OF GENETIC DAMAGE IN HUMAN CELLS

白血病和淋巴瘤各新增8万多例的原因 在美国，人们基本上不知道这一年的情况。一簇簇的白血病已经 发现在超级基金地点和其他危险设施附近，筹集公众资金 对环境化学品所起作用的担忧。需要新的方法来 确定哪些危险因素在白血病的发生和发展中起作用 预测哪些化学物质可能导致白血病。在过去的几年里，许多人 与白血病相关的重要基因变化中 已确认身份。此外，人们已经认识到，这些变化必须发生在 存在于骨髓中的早期祖细胞或干细胞。因此， 有可能导致白血病的化学物质应该进行检测 在培养的细胞中产生与白血病相关的基因变化的能力 干细胞。幸运的是，茎的分离和培养方法 最近从外周血中提取的细胞已经被开发出来。此外, 可检测与白血病相关的重要基因变化的方法 也被开发出来了。这些方法是基于原位荧光的。 杂交(FISH)和逆转录-聚合酶链式反应(RT-PCR)。我们建议 使用这些方法和微核试验来确定是否选择了 化学物质，包括那些在超级基金网站发现的化学物质，可以产生基因 培养的淋巴细胞和祖细胞与白血病的相关性损伤 (干细胞)培养中的细胞。此外，我们计划使用FISH和RT-PCR来 描述200例儿童新病例中发生的基因变化 白血病与项目6。这将允许对 因此，可以观察到与化学物质暴露有新的联系。 尤其是。特别是，各种疾病的发生时间和流行程度 染色体易位将被确定。 最近的证据表明，发生了大量这样的移位 由于V(D)J重组酶活性异常。因此，我们建议 确定V(D)J重组酶活性是否可被诱导 精选的化学品。我们还将确定儿童病例是否 白血病V(D)J重组酶介导的异常事件水平较高 在他们的外周血液中。这将提供对该角色的进一步洞察 V(D)J重组酶活性异常在白血病发生中的作用我们也 建议检验B淋巴细胞可能是更好的替代品的假设 T细胞在人类人群的生物标志物流行病学研究中的作用。我们 将测试各种化学物质在B-细胞中产生遗传损伤的能力 和T淋巴细胞直接比较产生的数据。此外，我们还将 确定基因多态性在谷胱甘肽转移酶中的作用 人类对化学诱导的遗传损伤易感性的同工酶(in 与项目4的协作)。因此，新的分析方法将被开发成 更好地预测哪些化学物质可能导致人类白血病。在……里面 此外，与项目4和6的调查人员一起，我们将 确定哪些因素最有可能导致白血病的发生， 一种通常聚集在超级基金网站附近的疾病。

The causes of the more than 80,000 new cases of leukemia and lymphoma each year in the US are largely unknown. "Clusters" of leukemias have been found near Superfund sites and other hazardous facilities, raising public concern over the role environmental chemicals. New methods are needed to determine what risk factors play a role in leukemia development and to predict which chemicals might cause leukemia. In the past few years, many of the important genetic changes associated with leukemia have been identified. Further, it has been realized that these changes must occur in early progenitor or stem cells residing in the bone marrow. Hence, chemicals with the potential to cause leukemia should be tested for their ability to produce genetic changes associated with leukemia in cultured stem cells. Fortunately, methods for the isolation and culture of stem cells from peripheral blood have recently been developed. In addition, methods which can detect important genetic changes associated with leukemia have also been developed. These methods are based on fluorescence in situ hybridization (FISH) and reverse transcriptase - PCR (rt-PCR). We propose to use these methods and the micronucleus assay to determine if selected chemicals, including those found at Superfund sites, can produce genetic damage of relevance to leukemia in cultured lymphocytes and progenitor (stem) cells in culture. Further, we plan to use FISH and rt-PCR to characterize the genetic changes occurring in 200 new cases of childhood leukemia with Project 6. This will allow for subclassification of the leukemias so that new associations with chemical exposures can be observed. In particular. In particular, the timing and prevalence of various chromosomal translocations will be determined. Recent evidence suggests that a large number of these translocations occur as a result of aberrant V (D)J recombinase activity. We therefore propose to determine if aberrant V(D)J recombinase activity can be induced by selected chemicals. We will also determine if the cases of childhood leukemia have a higher level of aberrant V(D)J recombinase-mediated events in their peripheral blood. This will provide further insight into the role of aberrant V(D)J recombinase activity in producing leukemia. We also propose to test the hypothesis that B-lymphocytes may be better surrogates than T-cells in biomarker epidemiology studies of human populations. We will test the ability of various chemicals to produce genetic damage in B- and T-lymphocytes and directly compare the data produced. Further, we will determine the role of genetic polymorphisms in glutathione transferase isoenzymes in human susceptibility to chemically induced genetic damage (in collaboration with Project 4). Thus, new assays will be developed to better predict which chemicals are likely to cause human leukemia. In addition, together with investigators from Projects 4 and 6, we will determine what factors most likely contribute the development of leukemia, a disease commonly clustering near Superfund sites.