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一种使用成像流细胞计的多生物标志物微核试验.

Danielle S G Harte^1,2, Anthony M Lynch^1,2, Jatin Verma¹

¹Swansea University Medical School, Swansea University, Swansea, UK.

Archives of toxicology

|July 12, 2024

概括

这项研究提出了一种新的成像流细胞计方法,用于评估化学基因毒性,减少药物开发中的错误阳性. 该协议准确地识别了DNA损伤和作用模式,改善了早期的候选药物选择,并将动物试验降到最低.

关键词:

生物标志物生物标志物造成的DNA损伤是DNA损伤.图像流 (ImageStream) 是一个这是一个微核.这是一个MoA MoA.这个名字是NAM NAM.

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科学领域:

毒理学毒理学毒理学
分子生物学分子生物学
细胞生物学细胞生物学

背景情况:

在体外遗传毒性测试往往产生误导性的结果,导致不必要的动物试验和早期拒绝有前途的候选药物.
了解化学作用模式 (MoA) 对于准确评估基因毒性潜力和临床风险至关重要.
目前的方法可以与高的假阳性率作斗争,需要改进查试验.

研究的目的:

使用成像流细胞计测试开发一种强大,高通量的基因毒性测试协议.
为了能够同时评估DNA损伤生物标志物和未解细胞中的微核 (MN) 形成.
为了更精确地区分克拉斯托基因和无基因化学MOA,以改善药物开发查.

主要方法:

开发了用于固定人体淋巴细胞TK6细胞的多重染色协议,使用抗体对抗hH2AX,p53,pH3S28和DRAQ5TMDNA染色.
使用了Cytek® Amnis® ImageStream®X Mk II成像流动细胞计,用于高通量数据采集.
在 IDEAS® 6.2 软件中实现了自动化掩护和封闭策略,用于量化细胞周期,MN和生物标记物种群.

主要成果:

成功演示了一种多重系统,用于DNA损伤评估和在未被解读的细胞中识别MN.
开发的门关策略可以实现自动批处理和同时量化多个细胞事件.
用卡本达和甲基甲硫酸盐 (MMS) 进行的概念验证实验准确地确定了它们各自的克拉斯托基因和无菌原性MoAs.

结论:

本次展示的成像流细胞测量方案为基因毒性测试提供了强大而高效的方法.
这种多重方法提高了基因毒性评估的准确性,有助于识别更安全的候选药物.
能够确定化学MoA的能力直接有助于减少假阳性和优化药物开发管道.