Investigating why Cdk4/6 inhibitors have durable long-term effect on tumour growth.

这项提案将阐述在治疗转移性乳腺癌方面显示出显著效果的CDK4/6抑制剂如何导致持久的长期细胞周期停滞。我们将使用细胞周期报告器进行活细胞成像，以准确测量单个细胞中的药物反应。我们在健康的二倍体细胞中的初步工作表明，它们在最初几天可逆地停滞，然后进入不可逆的细胞周期停滞状态。我们将研究在可逆性中控制这一开关的分子途径，我们还将确定它在对CDK4/6抑制反应更好的癌细胞系中是否被更快地触发。最后，我们将使用表型筛选来寻找新的方法来改善CDK4/6抑制后的长期停滞。预测结果预期这些研究将有助于解释：1)为什么CDK4/6抑制剂在临床上如此有效；2)哪些肿瘤基因类型将从CDK4/6抑制中受益(即帮助开发一种精确的药物策略)；以及3)什么样的联合治疗可以对CDK4/6抑制产生持久的长期反应。培训这个翻译项目将允许学生在分子和细胞生物学、高含量显微镜与定量计算分析以及表型药物筛选之间的界面上接受跨学科培训。问题：1.解释跨学科接口：将开发定量方法，以允许在单个细胞中进行自动化的细胞周期分析。这将被用作翻译表型筛选计划的基础，以寻找组合策略来改善CDK4/6抑制剂的长期效果。学生还将使用各种分子和细胞生物学技能来分析途径，并确定对CDK4/6抑制的反应。项目是否需要大量的量化技能？YES3.项目是否需要大量的整体生理技能？不是

This proposal will address how Cdk4/6 inhibitors, which have shown remarkable efficacy at treating metastatic breast cancer, cause a durable long-term cell cycle arrest. We will perform live-cell imaging with cell cycle reporters to accurately measure drug-responses in individual cells. Our preliminary work in healthy diploid cells, demonstrates that they arrest reversibly over the first few days before withdrawing into a state of irreversible cell cycle arrest. We will investigate the molecular pathways that control this switch in reversibility and we will also determine whether it is triggered quicker in cancer cell lines that respond better to Cdk4/6 inhibition. Finally, we will use phenotypic screening to search for novel ways to improve long-term arrest following Cdk4/6 inhibition. Predicted OutcomesThe expectation is that these studies will help to explain: 1) why Cdk4/6 inhibitors are so effective in the clinic; 2) what tumour genotypes will benefit from Cdk4/6 inhibition (i.e. to help develop a precision medicine strategy); and 3) what combinations treatments can produce a durable long-term response to Cdk4/6 inhibition. TrainingThis translational project will allow the student to receive interdisciplinary training at the interface between molecular and cellular biology, high-content microscopy with quantitative computational analysis, and phenotypic drug screening. Questions:1. Explain interdisciplinary interface: Quantitative approaches will be developed to allow automated cell cycle analysis in individual cells. This will be used as a basis for a translational phenotypic screening program to find combination strategies to improve the long-term effects of Cdk4/6 inhibitors. Student will also use a variety of molecular and cell biology skills to analyse the pathways the determine the response to Cdk4/6 inhibition.2. Does project require significant amount of quantitative skills? YES3. Does project require significant amount of whole organism physiology skills? NO