In vitro teratogenicity testing using a 3D, embryo-like gastruloid system.
In vitro teratogenicity testing using a 3D, embryo-like gastruloid system.
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DOI：
10.1016/j.reprotox.2021.08.003
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发表时间：
2021-10
期刊：
影响因子：
--
通讯作者：
Moris N
中科院分区：
文献类型：
--
作者：
Mantziou V;Baillie-Benson P;Jaklin M;Kustermann S;Arias AM;Moris N
类胃泌素是一种新的致畸性体外试验平台。致畸剂破坏胃类基因表达和形态与成像读出。中等通量胃类细胞培养物可定量，具有统计学稳健性。小鼠和人类类胃体细胞对致畸剂的敏感性。作为具有自动化范围的预测性测定的概念验证。预期用于有生育能力患者的药物在上市前需要进行致畸性试验。几家制药公司使用无动物体外模型，允许更快速地选择先导化合物，并通过简化提交给动物进一步监管研究的有前途化合物的选择，为3Rs原则（“替换、减少和改进”）做出贡献。目前可用的体外模型通常依赖于贴壁单层培养物或无序的3D结构，这两者都缺乏发育胚胎的时空和形态背景。一个新开发的3D 'gastruloid'模型有可能实现一个更可靠的预测致畸性提供了一个强大的重演原肠胚样事件旁边的形态协调在相对高的吞吐量。在这第一个概念验证研究中，我们使用小鼠和人类类胃腺激素检查了一组7种参比化合物，以及相关的体内数据和已知的致畸风险，以定量评估体外致畸性。在暴露于几种参比化合物后，我们观察到几种大体形态学效应，包括显著降低的伸长或减小的胃状体大小。我们还观察到异常的基因表达使用荧光报告，包括SOX 2，BRA，和SOX 17，提示多谱系分化缺陷和破坏轴向图案。最后，我们看到，gastruloids概括了一些已知的体内物种特异性的亲和性之间的小鼠和人类的同行。因此，我们认为，gastruloids是一个强大的工具，致畸性评估，使相关的生理重演早期胚胎发育，证明其作为一种新的体外致畸模型系统的使用。
Gastruloids are a new in vitro platform for teratogenicity testing. Teratogens disrupt gastruloid gene expression and morphology with imaging readout. Medium-throughput gastruloid cultures are quantifiable with statistical robustness. Mouse and human gastruloids recapitulate species-specific sensitivities to teratogens. Proof-of-concept as a predictive assay with scope for automation. Pharmaceuticals intended for use in patients of childbearing potential need to be tested for teratogenicity before marketing. Several pharmaceutical companies use animal-free in vitro models which allow a more rapid selection of lead compounds and contribute to 3Rs principles (‘replace, reduce and refine’) by streamlining the selection of promising compounds submitted to further regulatory studies in animals. Currently available in vitro models typically rely on adherent monolayer cultures or disorganized 3D structures, both of which lack the spatiotemporal and morphological context of the developing embryo. A newly developed 3D ‘gastruloid’ model has the potential to achieve a more reliable prediction of teratogenicity by providing a robust recapitulation of gastrulation-like events alongside morphological coordination at relatively high-throughput. In this first proof-of-concept study, we used both mouse and human gastruloids to examine a panel of seven reference compounds, with associated in vivo data and known teratogenic risk, to quantitatively assess in vitro teratogenicity. We observed several gross morphological effects, including significantly reduced elongation or decreased size of the gastruloids, upon exposure to several of the reference compounds. We also observed aberrant gene expression using fluorescent reporters, including SOX2, BRA, and SOX17, suggestive of multi-lineage differentiation defects and disrupted axial patterning. Finally, we saw that gastruloids recapitulated some of the known in vivo species-specific susceptibilities between their mouse and human counterparts. We therefore suggest that gastruloids represent a powerful tool for teratogenicity assessment by enabling relevant physiological recapitulation of early embryonic development, demonstrating their use as a novel in vitro teratogenic model system.
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