Department of Safety Assessment, 709 Swedeland Road, King of Prussia, PA 19406, USA.
Toxicol Appl Pharmacol. 2011 Dec 1;257(2):284-300. doi: 10.1016/j.taap.2011.09.015. Epub 2011 Sep 24.
Drug-induced vascular injury is frequently observed in rats but the relevance and translation to humans present a hurdle for drug development. Numerous structurally diverse pharmacologic agents have been shown to induce mesenteric arterial medial necrosis in rats, but no consistent biomarkers have been identified. To address this need, a novel strategy was developed in rats to identify genes associated with the development of drug-induced mesenteric arterial medial necrosis. Separate groups (n=6/group) of male rats were given 28 different toxicants (30 different treatments) for 1 or 4 days with each toxicant given at 3 different doses (low, mid and high) plus corresponding vehicle (912 total rats). Mesentery was collected, frozen and endothelial and vascular smooth muscle cells were microdissected from each artery. RNA was isolated, amplified and Affymetrix GeneChip® analysis was performed on selectively enriched samples and a novel panel of genes representing those which showed a dose responsive pattern for all treatments in which mesenteric arterial medial necrosis was histologically observed, was developed and verified in individual endothelial cell- and vascular smooth muscle cell-enriched samples. Data were confirmed in samples containing mesentery using quantitative real-time RT-PCR (TaqMan™) gene expression profiling. In addition, the performance of the panel was also confirmed using similarly collected samples obtained from a timecourse study in rats given a well established vascular toxicant (Fenoldopam). Although further validation is still required, a novel gene panel has been developed that represents a strategic opportunity that can potentially be used to help predict the occurrence of drug-induced mesenteric arterial medial necrosis in rats at an early stage in drug development.
药物引起的血管损伤在大鼠中经常观察到，但与人类的相关性和转化存在药物开发的障碍。许多结构不同的药理制剂已被证明可在大鼠中诱导肠系膜动脉中层坏死，但尚未确定一致的生物标志物。为了解决这一需求，在大鼠中开发了一种新策略，以鉴定与药物诱导的肠系膜动脉中层坏死发展相关的基因。将雄性大鼠分成几组（每组 6 只），分别给予 28 种不同的毒物（30 种不同的处理），每种毒物以 3 种不同剂量（低、中、高）加相应的载体（912 只大鼠）给药 1 或 4 天。收集肠系膜，冷冻，从每条动脉中分离出内皮细胞和血管平滑肌细胞。分离出 RNA，进行扩增，并对选择性富集的样本进行 Affymetrix GeneChip®分析，并开发了一个新的基因表达谱，代表了在所有治疗中观察到组织学上的肠系膜动脉中层坏死的所有处理中均表现出剂量反应模式的基因，并在单独的内皮细胞和血管平滑肌细胞富集样本中进行了验证。使用定量实时 RT-PCR（TaqMan™）基因表达谱在含有肠系膜的样本中确认了数据。此外，还使用从给予既定血管毒物（Fenoldopam）的大鼠时间过程研究中收集的类似样本确认了该基因表达谱的性能。尽管仍需要进一步验证，但已经开发出一种新的基因表达谱，这代表了一种战略机会，可潜在地用于帮助预测药物诱导的大鼠肠系膜动脉中层坏死的发生，从而在药物开发的早期阶段。
