Department of Genetics, Evolution and Environment, Centre for Life's Origins and Evolution, University College London, London WC1E 6BT, UK.
Syngenta, Jealott's Hill International Research Centre, Bracknell RG42 6EY, UK.
Toxicol Sci. 2021 Sep 28;183(2):302-318. doi: 10.1093/toxsci/kfab097.
A critical aspect of toxicity evaluation is developmental and reproductive toxicity (DART) testing. Traditionally, DART testing has been conducted in vivo in mammalian model systems. New legislation aimed at reducing animal use and the prohibitive costs associated with DART testing, together with a need to understand the genetic pathways underlying developmental toxicity means there is a growing demand for alternative model systems for toxicity evaluation. Here we explore the potential of the eukaryotic social amoeba Dictyostelium discoideum, which is already widely used as a simple model system for cell and developmental biology, as a potential nonanimal model for DART testing. We developed assays for high-throughput screening of toxicity during D. discoideum growth and development. This allowed the toxicity of a broad range of test compounds to be characterized, which revealed that D. discoideum can broadly predict mammalian toxicity. In addition, we show that this system can be used to perform functional genomic screens to compare the molecular modes of action of different compounds. For example, genome-wide screens for mutations that affect lithium and valproic acid toxicity allowed common and unique biological targets and molecular processes mediating their toxicity to be identified. These studies illustrate that D. discoideum could represent a predictive nonanimal model for DART testing due to its amenability to high-throughput approaches and molecular genetic tractability.
毒性评估的一个关键方面是发育和生殖毒性(DART)测试。传统上,DART 测试是在哺乳动物模型系统中进行的体内实验。新的立法旨在减少动物使用量,并降低与 DART 测试相关的高昂成本,同时需要了解发育毒性的遗传途径,这意味着人们对毒性评估的替代模型系统的需求日益增长。在这里,我们探讨了真核生物有丝分裂变形虫(Dictyostelium discoideum)的潜力,它已经被广泛用作细胞和发育生物学的简单模型系统,作为 DART 测试的潜在非动物模型。我们开发了用于高通量筛选 D. discoideum 生长和发育过程中毒性的测定法。这使得可以对广泛的测试化合物的毒性进行特征描述,结果表明 D. discoideum 可以广泛预测哺乳动物的毒性。此外,我们还表明,该系统可用于进行功能基因组筛选,以比较不同化合物的分子作用模式。例如,针对影响锂和丙戊酸毒性的突变的全基因组筛选,使我们能够鉴定共同和独特的生物靶标以及介导其毒性的分子过程。这些研究表明,由于 D. discoideum 易于进行高通量方法和分子遗传学研究,因此它可能代表用于 DART 测试的有预测性的非动物模型。