一种新的体外人类骨髓三维模型,以弥合体外和体内遗传毒性测试之间的差距。

IF 2.5 4区 医学 Q3 GENETICS & HEREDITY
Mutagenesis Pub Date : 2022-05-04 DOI:10.1093/mutage/geac009
Alexander R Vernon,Roy M Pemberton,H Ruth Morse
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引用次数: 2

摘要

调控2D体外微核(MN)测定是一系列测试的一部分,用于在体内评估新化合物和现有化合物之前测试其遗传毒性(ICH S2)。二维骨髓分析包括单层细胞,而体内骨髓(BM)环境包括三维细胞外基质内的多细胞环境。尽管体外MN测定遵循经济合作与发展组织(OECD)制定的严格协议,以符合监管机构的要求,但一些化合物在体外MN测定中被确定为阴性基因毒物,但在体内评估时却略微呈阳性。糖皮质激素在体内呈弱阳性,一般认为不会造成长期致癌风险;然而,对于未知活性的新化合物,改进遗传毒性预测是必要的。为了帮助解决这一观察,我们描述了一种新的3D体外实验,旨在复制在体内BM微环境中看到的结果。对AlgiMatrix支架进行优化,以HS-5人骨髓基质细胞作为骨髓微环境,在基质中添加人淋巴母细胞TK6。根据2D调节试验方案进行MN试验。利用这种新型的BM体外3D模型,研究了已知的基因毒物(丝裂霉素C、依托泊苷和紫杉醇)、阴性对照(咖啡因)和体内阳性糖皮质激素(地塞米松和强的松龙)对MN的诱导作用。研究发现,与历史体内数据一致,该模型可以准确预测糖皮质激素的体内结果,而不像2D体外MN测定。这些初步结果表明,与标准的2D分析相比,我们的3D MN分析可以更好地预测体内MN测试的结果。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A novel in vitro 3D model of the human bone marrow to bridge the gap between in vitro and in vivo genotoxicity testing.
The regulatory 2D in vitro micronucleus (MN) assay is part of a battery of tests, used to test for genotoxicity of new and existing compounds before they are assessed in vivo (ICH S2). The 2D MN assay consists of a monolayer of cells, whereas the in vivo bone marrow (BM) setting comprises a multicellular environment within a three-dimensional extracellular matrix. Although the in vitro MN assay follows a robust protocol set out by the Organisation for Economic Co-operation and Development (OECD) to comply with regulatory bodies, some compounds have been identified as negative genotoxicants within the in vitro MN assay but marginally positive when assessed in vivo. The glucocorticoids, which are weakly positive in vivo, have generally been suggested to pose no long-term carcinogenic risk; however, for novel compounds of unknown activity, improved prediction of genotoxicity is imperative. To help address this observation, we describe a novel 3D in vitro assay which aims to replicate the results seen within the in vivo BM microenvironment. AlgiMatrix scaffolds were optimized for seeding with HS-5 human BM stromal cells as a BM microenvironment, to which the human lymphoblast cell line TK6 was added. An MN assay was performed aligning with the 2D regulatory assay protocol. Utilizing this novel 3D in vitro model of the BM, known genotoxicants (mitomycin C, etoposide, and paclitaxel), a negative control (caffeine), and in vivo positive glucocorticoids (dexamethasone and prednisolone) were investigated for the induction of MN. It was found, in agreement with historical in vivo data, that the model could accurately predict the in vivo outcome of the glucocorticoids, unlike the regulatory 2D in vitro MN assay. These preliminary results suggest our 3D MN assay may better predict the outcome of in vivo MN tests, compared with the standard 2D assay.
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来源期刊
Mutagenesis
Mutagenesis 生物-毒理学
CiteScore
5.90
自引率
3.70%
发文量
22
审稿时长
6-12 weeks
期刊介绍: Mutagenesis is an international multi-disciplinary journal designed to bring together research aimed at the identification, characterization and elucidation of the mechanisms of action of physical, chemical and biological agents capable of producing genetic change in living organisms and the study of the consequences of such changes.
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