Mechanistic Pharmacology of Anti-Mitotics and Apoptosis Regulation

描述（由申请人提供）：本项目的长期目标是以精确的定量方式了解单个癌细胞和肿瘤对药物治疗的反应，从靶向作用到诱导细胞凋亡到最终肿瘤消退。这将通过改进药物反应预测和合理设计联合疗法，以及通过确定未来更好药物的靶标，改善患者护理。我们将在两种触发癌细胞凋亡的药物、抗有丝分裂药物和靶向细胞凋亡诱导剂（包括TRAIL和ABT737）的背景下解决这一目标。实验将在细胞培养和小鼠肿瘤中进行。我们的目标是理解细胞对这些药物的反应，这是(i)在解释特定蛋白质和其他生物分子之间相互作用的细胞表型方面的机制；（ii）定量应用质量作用动力学和其他数学形式来预测相互作用蛋白质的行为，从他们的个体生物化学知识；（iii）概率在解释从一个细胞到下一个细胞对药物反应的可变性与此同时，只有一小部分肿瘤细胞在化疗药物治疗后可能会停止或死亡（iv）后基因组分析不同细胞系（最终是患者样本），了解它们的遗传差异，并有可能应用强大的敲除/入和RNAi策略来改变基因型(v)整合假设药物反应的决定因素是多因素的，并且有多种相互作用的途径，而不是单一的必须研究单个基因或蛋白质。在目标1中，我们将确定在抗有丝分裂药物和ABT737的作用下调节MOMP的分子机制。在目标2中，我们将研究细胞对抗有丝分裂和靶向凋亡诱导剂反应变化的原因。在目标3中，我们将询问在细胞培养和小鼠肿瘤中，使用活体成像和其他方法，药物反应在多大程度上相同。在目标4中，我们将采用几种方法将目标1-3的机制理解转化为改善患者护理。

DESCRIPTION (provided by applicant): The long-term goal of this Program Project is to understand, in precise quantitative terms, how individual cancer cells and tumors respond to drug treatment, from target engagement to induction of apoptosis to eventual tumor regression. This will improve patient care by allowing improved prediction of drug responses and rational design of combination therapies, and by identifying targets for better future drugs. We will address this goal in the context of two drug classes that trigger apoptosis in cancer cells, anti-mitotic drugs, and targeted apoptosis inducers, including TRAIL and ABT737. Experiments will be performed in cell culture and mouse tumors. We aim for an understanding of the cellular response to these drugs that is (i) mechanistic in explaining cellular phenotypes in terms of interactions among specific proteins and other bio-molecules (ii) quantitative in applying mass-action kinetics and other mathematical formalisms to predicting the behavior of ensembles of interacting proteins from knowledge of their individual biochemistry (iii) probabilistic in accounting for the variability from one cell to the next in responses to drugs with the attendant likelihood that only a fraction of tumor cells will arrest or die in response to treatment with a chemotherapeutic drug (iv) post-genomic in analyzing diverse cell lines (and ultimately patient samples) with knowledge of their genetic differences and with the possibility of applying powerful knock-out/in and RNAi strategies to alter genotype (v) integrative in assuming that determinants of drug response are multi-factorial and that multiple interacting pathways rather than single genes or proteins must be studied. We will address these goals in four Program Specific Aims: In aim 1 we will determine the molecular mechanisms that regulate MOMP in response to anti-mitotic drugs and ABT737. In aim 2 we will investigate the causes of variation in cell responses to anti-mitotics and targeted inducers of apoptosis. In aim 3 we will ask to what extent drug responses are the same in cell culture and mouse tumors, using intravital imaging and other methods. In aim 4 we will pursue several approaches towards translating mechanistic understanding from aims 1-3 into improved patient care.