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Erica Coratella¹, Rebecca Bohnert², Benoit Fischer²

¹Novartis Biomedical Research, Basel, Switzerland; Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, 4056 Basel, Switzerland.

Toxicology and applied pharmacology

|August 23, 2025

概括

在低附着度测试中,小鼠软菌形成和生长对肝细胞癌 (HCC) 发展的预测值较低. 这些测试未能检测出与重组腺相关病毒 (rAAV) 和瘤抑制剂相关的瘤性.

科学领域:

肝胆细胞瘤
分子瘤学
基因毒性检测

背景情况:

在新生小鼠中,重组腺相关病毒 (rAAV) 集成与肝细胞癌 (HCC) 有关.
由于缺乏合适的体外模型,对患者的癌症风险进行评估是具有挑战性的.
为评估瘤性,建议进行软菌殖民地形成 (SACF) 和低附着增长 (GILA) 等独立于基的生长测试.

研究的目的:

开发和验证SACF和GILA测试的小鼠版本,以评估HCC的发展风险.
在体外评估基因组编辑和rAAV诱导的插入突变的致癌潜力.
使用CRISPR/Cas9技术研究特定瘤抑制基因在HCC发育中的作用.

主要方法:

在小鼠肝细胞系AML12中,选择的瘤抑制剂 (Nf2, Tp53, Pten, Axin1, Ctnnb1, Tsc1) 通过CRISPR/ Cas9进行淘汰.
产生稳定的AML12和人类MCF10A细胞克隆,分别将rAAV基因组集成到Rian和MEG8位点中.
通过小鼠SACF和GILA测定,评估结独立的生长.

主要成果:

在AML12细胞中诱导了Nf2的淘汰和Tp53/Pten的双重失活.
编辑Axin1,Ctnnb1和Tsc1并没有促进结独立的生长.
在SACF和GILA测定中,具有集成rAAV基因组的AML12和MCF10A克隆没有表现出结独立性.

结论:

小鼠SACF和GILA测定对HCC发育具有有限的预测价值.
这些测试未能检测出与rAAV整合和瘤抑制基因失活相关的瘤性.
需要开发更准确的体外方法来评估rAAV的基因毒性和致癌风险.

关键词:

腺相关病毒安克雷奇独立增长肝细胞癌在体外转化试验插入性瘤发生抑制瘤的药物

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