Genome instability is a hallmark of most human cancers and is exacerbated following replication stress. However, the effects that drugs/xenobiotics have in promoting genome instability including chromosomal structural rearrangements in normal cells are not currently assessed in the genetic toxicology battery. Here, we show that drug‐induced replication stress leads to increased genome instability in vitro using proliferating primary human cells as well as in vivo in rat bone marrow (BM) and duodenum (DD). p53‐binding protein 1 (53BP1, biomarker of DNA damage repair) nuclear bodies were increased in a dose‐dependent manner in normal proliferating human mammary epithelial fibroblasts following treatment with compounds traditionally classified as either genotoxic (hydralazine) and nongenotoxic (low‐dose aphidicolin, duvelisib, idelalisib, and amiodarone). Comparatively, no increases in 53BP1 nuclear bodies were observed in nonproliferating cells. Negative control compounds (mannitol, alosteron, diclofenac, and zonisamide) not associated with cancer risk did not induce 53BP1 nuclear bodies in any cell type. Finally, we studied the in vivo genomic consequences of drug‐induced replication stress in rats treated with 10 mg/kg of cyclophosphamide for up to 14 days followed by polymerase chain reaction‐free whole genome sequencing (30X coverage) of BM and DD cells. Cyclophosphamide induced chromosomal structural rearrangements at an average of 90 genes, including 40 interchromosomal/intrachromosomal translocations, within 2 days of treatment. Collectively, these data demonstrate that this drug‐induced genome instability test (DiGIT) can reveal potential adverse effects of drugs not otherwise informed by standard genetic toxicology testing batteries. These efforts are aligned with the food and drug administration's (FDA's) predictive toxicology roadmap initiative.

中文翻译：

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药物诱导的复制应激后正常哺乳动物基因组中基因组不稳定的生物标志物。

基因组不稳定性是大多数人类癌症的标志，在复制压力下加剧。但是，目前尚不能在遗传毒理学领域评估药物/异种生物对促进基因组不稳定性（包括正常细胞中的染色体结构重排）的影响。在这里，我们显示了药物诱导的复制应激导致使用增殖的原代人细胞以及大鼠骨髓（BM）和十二指肠（DD）体内的基因组不稳定性增加。用传统分类为遗传毒性（肼苯哒嗪）和非遗传毒性（低剂量的蚜虫碱）的化合物处理后，正常增殖的人类乳腺上皮成纤维细胞中，p53结合蛋白1（53BP1，DNA损伤修复的生物标记）的核体呈剂量依赖性增加。 ，duvelisib，idelalisib，和胺碘酮）。比较而言，在非增殖细胞中未观察到53BP1核体的增加。与癌症风险无关的阴性对照化合物（甘露醇，醛固酮，双氯芬酸和唑尼沙胺）在任何细胞类型中均未诱导出53BP1核体。最后，我们研究了在用10 mg / kg环磷酰胺处理长达14天，然后进行无聚合酶链反应的BM和DD细胞全基因组测序（覆盖30倍）的大鼠中，药物诱导的复制应激的体内基因组后果。在治疗后2天内，环磷酰胺平均可诱导90个基因发生染色体结构重排，包括40个染色体间/染色体内易位。总的来说，这些数据表明，这种药物诱导的基因组不稳定性测试（DiGIT）可以揭示标准遗传毒理学测试电池无法告知的药物的潜在不良反应。这些努力与食品和药物管理局（FDA）的预测毒理学路线图计划相一致。