Evans Stephen J, Gollapudi Bhaskar, Moore Martha M, Doak Shareen H
In Vitro Toxicology Group, Institute of Life Science, Swansea University Medical School, Swansea University, Singleton Park, Swansea SA2 8PP, Wales, UK.
Exponent, Inc., 1800 Diagonal Road, Suite 500, Alexandria, VA 22314, USA.
Mutat Res Genet Toxicol Environ Mutagen. 2019 Nov;847:403024. doi: 10.1016/j.mrgentox.2019.02.005. Epub 2019 Feb 21.
The induction of gene mutation within a DNA sequence can result in an adverse impact, altering or preventing gene function. Therefore, in vitro evaluation of mutagenicity is an essential component of the toxicological screening process. A variety of mutagen screening tools are routinely used in genetic toxicology, which are based on selected reporter genes. These assays are however typically labour intensive and impractical for high throughput screening. Considering this, the IWGT (International Workshops on Genotoxicity Testing) sub-group on Novel & Emerging in vitro Mammalian Cell Mutagenicity Test Systems undertook a literature search to identify new approaches for mutation detection. This review therefore focused on identifying new approaches for mutation detection that have the potential for use as a future genotoxicity screening tool. A comprehensive literature review identified genome-wide loss-of-function screening tools, next generation sequencing (NGS) mutation characterisation and fluorescence-based mutation detection methods as having significant promise as an emerging in vitro mammalian cell mutagenicity test system. Each of the technologies considered was assessed for its capacity to report on a wide array of heritable mutagenic changes, necessary to cover the full spectrum of genetic events imparted by substances with a broad range of modes of action. Of the technologies evaluated, NGS techniques exhibited the greatest advantages for use in a genotoxicity testing setting. However, it is important to note that the emerging techniques identified could not facilitate routine mutagenicity testing in their current format and require substantial additional optimisation and tailoring before they could be utilised as an in vitro mammalian cell mutagenicity test system. Additionally, new mammalian cell mutation test systems must be able to accurately and reliably detect and quantify rare events; hence any new system would require careful validation. Nevertheless, with further development emerging technologies such as NGS could become important in establishing more predictive and high-throughput regulatory hazard screening tools of the future.
DNA序列内基因突变的诱导可导致不良影响，改变或阻止基因功能。因此，体外致突变性评估是毒理学筛选过程的重要组成部分。遗传毒理学中常规使用多种基于选定报告基因的诱变筛选工具。然而，这些检测通常劳动强度大，不适用于高通量筛选。考虑到这一点，国际遗传毒性测试研讨会（IWGT）新型和新兴体外哺乳动物细胞致突变性测试系统小组进行了文献检索，以确定新的突变检测方法。因此，本综述着重于确定具有用作未来遗传毒性筛选工具潜力的新的突变检测方法。全面的文献综述确定全基因组功能丧失筛选工具、下一代测序（NGS）突变表征和基于荧光的突变检测方法作为新兴的体外哺乳动物细胞致突变性测试系统具有重大前景。所考虑的每项技术都根据其报告广泛的可遗传诱变变化的能力进行了评估，这些变化对于涵盖具有广泛作用模式的物质所引发的全部遗传事件谱是必要的。在所评估的技术中，NGS技术在遗传毒性测试环境中使用表现出最大优势。然而，重要的是要注意，所确定的新兴技术目前的形式无法促进常规致突变性测试，在能够用作体外哺乳动物细胞致突变性测试系统之前需要大量额外的优化和调整。此外，新的哺乳动物细胞突变测试系统必须能够准确可靠地检测和量化罕见事件；因此，任何新系统都需要仔细验证。尽管如此，随着进一步发展，诸如NGS之类的新兴技术在建立未来更具预测性和高通量的监管危害筛选工具方面可能会变得很重要。
