Quantitative Analysis of Cell Death Pathways in Cancer

这个由Peter Sorger教授领导的项目的总体目标是用精确的分子术语描绘 哺乳动物细胞暴露于小分子后启动凋亡的调控机制 和生物疗法。从一种细胞类型到另一种细胞类型以及从一个细胞到下一个细胞的差异将是 特别关注，最终目标是开发预测患者对治疗的特定反应的方法。 作为一种建立对细胞凋亡的机械性、概率性、整体性和预测性理解的手段，我们将 收集种群水平和单细胞的caspase激活动力学数据，并构建、校准和 分析描述哺乳动物细胞死亡的关键步骤的一系列数学模型。我们将专注于 主要是现有的和研究中的抗癌药物，但也希望检查改变 炎症和免疫反应。我们选择治疗药物的指导原则是：(I) 常规药物在癌症治疗中的标准护理和开发改进临床的可能性 方案(如紫杉烷)(二)患者之间药物反应的差异程度及其带来的困难 确定可能从特定治疗中受益的患者(III)新药物的潜力 显著改善临床前和临床研究(例如ABT-737)所证明的结果。四 具体目标包括：(1)路径控制的预测性和机械性分析 配体对细胞线粒体外膜通透性和半胱氨酸天冬氨酸酶活性的影响 触发外源性细胞凋亡(2)直接比较细胞与细胞之间的变异时间和发生 克隆细胞群体成员之间和不同肿瘤细胞系之间的细胞死亡(3)实验 微管毒物紫杉醇和Bcl2诱导内源性细胞凋亡的模型驱动分析 抑制剂ABT737与化疗药物联合应用对多种癌细胞的单细胞分析 LINES(4)肿瘤细胞有丝分裂和凋亡活体成像方法的发展和应用 在小鼠体内原位注射。这些目标的成功不仅会对细胞凋亡的研究产生影响，而且会对整个研究产生影响。 肿瘤生物学与化疗药物的使用

The overall goal of this project, led by Prof. Peter Sorger, isto delineate, in precise molecular terms, the mechanisms that regulate the onset of apoptosis in mammalian cells following exposure to small molecule and biological therapeutics. Variation from one cell type to the next and one cell to the next will be an area of particular focus, with the eventual goal of developing means to predict patient-specific responses to therapy. As a means to create mechanistic, probabilistic, integrative and predictive understanding of apoptosis we will collect population-level and single-cell data on caspase activation kinetics and construct, calibrate and analyze a series of mathematical models that describe key steps in mammalian cell death. We will focus largely on existing and investigational anti-cancer drugs, but also expect to examine agents that alter inflammatory and immune responses. Our selection of therapeutic agents is guided by (i) the importance of conventional agents in standard of care cancer treatment and the possibility of developing improved clinical protocols (e.g. taxanes) (ii) the extent of patient-patient variation in drug response and the attendant difficulty of identifying patients who might benefit from a particular treatment (iii) the potential of new agents to significantly improve outcomes as demonstrated by pre-clinical and clinical studies (e.g. ABT-737). Four specific aims will be pursued involving (1) predictive and mechanistic analysis of pathways controlling mitochondrial outer membrane permeablization and effector caspase activation in cells exposed to ligands that trigger extrinisic apoptosis (2) direct comparison of cell-to-cell variation in the timing and probability of cell death among members of a clonal cell population and between different tumor cell lines (3) experimental and model-driven analysis of intrinsic apoptosis induced by the microtubule poison paclitaxel and the Bcl2 inhibitor ABT737 and single-cell analysis of chemotherapeutics used in combination on diverse cancer cell lines (4) development and application of methods for intravital imaging of mitosis and apoptosis in cancer in situ in the mouse. Success with these aims will impact not only the study of apoptosis, but also general cancer biology and the use of chemotherapeutic drugs