Clinical Pharmacology and Safety Sciences, R&D, AstraZeneca, Cambridge, UK.
Sci Rep. 2021 Nov 9;11(1):21959. doi: 10.1038/s41598-021-01400-5.
Micronucleus (MN) assessment is a valuable tool in safety assessment. However, several compounds are positive in the in vivo bone marrow (BM) MN assay but negative in vitro, reflecting that BM complexity is not recapitulated in vitro. Importantly, these compounds are not genotoxic; rather, drug-driven pharmacological-effects on the BM increase MN, however, without mechanistic understanding, in vivo positives stop drug-progression. Thus, physiologically-relevant BM models are required to bridge the gap between in vitro and in vivo. The current study aimed to investigate the utility of two human 3D BM models (fluidic and static) for MN assessment. MN induction following treatment with etoposide and Poly-ADP Ribose Polymerase inhibitor (PARPi) and prednisolone (negative in vitro, positive in vivo) was determined in 2D L5178Y and human BM cells, and the 3D BM models. Etoposide (0-0.070 µM) and PARPi (0-150 µM) induced MN in both 3D BM models indicating their utility for genotoxicity testing. Interestingly, PARPi treatment induced a MN trend in 3D more comparable to in vivo. Importantly, prednisolone (0-1.7 mM) induced MN in both 3D BM models, suggesting recapitulation of the in vivo microenvironment. These models could provide a valuable tool to follow up, and eventually predict, suspected pharmacological mechanisms, thereby reducing animal studies.
微核(MN)评估是安全性评估的有力工具。然而,一些化合物在体内骨髓(BM)MN 测定中呈阳性,而在体外则呈阴性,这反映出 BM 的复杂性无法在体外重现。重要的是,这些化合物没有遗传毒性;相反,药物对 BM 的药理作用会增加 MN,但由于缺乏机制理解,体内阳性结果会阻止药物的进展。因此,需要具有生理相关性的 BM 模型来弥合体外和体内之间的差距。本研究旨在探讨两种人类 3D BM 模型(流体和静态)在 MN 评估中的应用。在 2D L5178Y 和人 BM 细胞以及 3D BM 模型中,测定了依托泊苷和聚二磷酸腺苷核糖聚合酶抑制剂(PARPi)和泼尼松龙(体外阴性,体内阳性)处理后 MN 的诱导情况。依托泊苷(0-0.070µM)和 PARPi(0-150µM)在两种 3D BM 模型中均诱导 MN,表明它们可用于遗传毒性测试。有趣的是,PARPi 处理在 3D 中诱导 MN 的趋势更类似于体内情况。重要的是,泼尼松龙(0-1.7mM)在两种 3D BM 模型中均诱导 MN,提示体内微环境的重现。这些模型可以为后续研究提供有价值的工具,最终预测可疑的药理机制,从而减少动物研究。