Does inappropriate DNA replication provide a mechanism for acquisition of drug resistance?

该项目将在PC 9细胞中获得对AZ药物Osimertinib（最佳EGFR抑制剂）的耐药性作为模型系统。i）我们将首先在奥希替尼治疗期间将scRNAseq应用于治疗期间的cRNA基因表达。单细胞方法将揭示单个复制细胞的状态，这可能是异质的，因为细胞在延长的治疗下不会均匀地反应。ii）我们已经开发了一种用于检测复制应激和DNA损伤的最先进的方法，TrAEL-seq，其以高灵敏度描绘DNA复制模式。TrAEL-seq将用于复制细胞，以了解药物治疗引起的复制应激。iii）复制应激可引起染色体外环状DNA（ecDNA），其经常含有赋予耐药性的基因扩增。将DTP中与药物相关的复制应激直接与ecDNA的形成联系起来将是特别令人兴奋的，我们将使用环状DNA测序进行测试。

This project will use the acquisition of resistance to the AZ drug Osimertinib, the best-in-class EGFR inhibitor, in PC9 cells as a model system. i) We will first apply scRNAseq during Osimertinib treatment to characterise gene expression during treatment. A single cell approach will reveal the status of individual replicating cells, which is likely to be heterogeneous as cells do not react uniformly under extended treatment.ii) We have developed a state-of-the-art method for detecting replication stress and DNA damage, TrAEL-seq, which profiles DNA replication patterns with high sensitivity. TrAEL-seq will be used to replicating cells to understand replication stress caused by drug treatment.iii) Replication stress can give rise to extrachromosomal circular DNA (ecDNA), which frequently harbours gene amplifications that bestow drug resistance. It would be particularly exciting to connect drug-assocatied replication stress in DTPs directly to formation of ecDNA, which we will test using circular DNA sequencing.