HIGH THROUGHPUT SYSTEM FOR MEASURING CYTOGENETIC DAMAGE

人类通过各种来源接触到基因毒性物质，包括工业过程、医疗、太阳辐射和生活方式因素，包括饮食和烟草使用。重要的是开发DNA损伤的生物标志物，以评估暴露、影响和在人类群体中的易感性。微核(MN)的形成是一个重要的遗传毒性终点，可用于有效地检测由断裂或非优生机制引起的DNA损伤。这种类型的染色体损伤与出生缺陷和癌症密切相关。一个灵敏和高通量的系统来测量人体血液样本中的MN将有无数的生物监测应用。例如，这样的系统可以提供关于正在进行临床试验的新药、化疗方案以及意外辐射或化学暴露的染色体破坏活性的重要信息。本项目基于流式细胞仪技术。第一阶段的目标是：1)招募受试者进行外周血采样；2)开发区分未成熟(RET)和成熟红细胞(RET)的程序；以及3)有效地计数RET中的微核。程序将基于Litron开发的一种自动化技术，用于检测啮齿动物血液样本中的染色体损伤。拟议的商业应用：这种方法的成功开发将使Litron实验室成为一家专家设施，能够对临床实验的人体血液样本进行细胞遗传学测量，并按服务收费。此外，通过将必要的试剂开发成试剂盒形式，将有可能将这项技术提供给其他能够使用单一激光流式细胞仪的实验室。这项技术创新技术有可能成为测量DNA损伤的重要临床工具。

Humans are exposed to genotoxic agents through a variety of sources including industrial processes, medical therapies, solar radiation and lifestyle factors including diet and tobacco use. It is important to develop biomarkers of DNA damage which evaluate exposure, effects and susceptibilities in human populations. Micronucleus (MN) formation is an important genotoxic endpoint which can be used to effectively detect DNA damage resulting from clastogenic or aneugenic mechanisms. Chromosomal damage of this type is closely associated with birth defects as well as cancer. A sensitive and high throughput system to measure human blood samples for MN would have a myriad of biomonitoring applications. For example, such a system could provide important information regarding the chromosome damaging activity of new drugs undergoing clinical trials, chemotherapy regimens, as well as accidental radiation or chemical exposures. This project is based on flow cytometry technology. The goals for Phase I are: 1) to recruit human subjects for peripheral blood sampling; 2) develop procedures to differentially label immature (RETs) and mature erythrocytes; and, 3) effectively enumerate micronuclei in RETs. Procedures will be based on an automated technique developed at Litron for detecting chromosome damage in rodent blood samples. PROPOSED COMMERCIAL APPLICATIONS: The successful development of this methodology will enable Litron Laboratories to become an expert facility, capable of performing cytogenetic measurements on human blood samples for clinical experiments on a fee- for-service basis. Additionally, by developing the necessary reagents into kit format, it would be possible to make this technology available to other laboratories having access to a single-laser flow cytometer. This technologically innovative technique has the potential to become an important clinical tool for measuring DNA damage.