Cell and Tumor Responses to Diverse Anti-Mitotic Drugs

细胞和肿瘤对多种抗有丝分裂药物的反应

• 批准号: 8375581
• 负责人: Timothy J Mitchison
• 金额: $ 31.18万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8375581
• 关键词: Address; Affect; Antimitotic Agents; Apoptosis; Apoptosis Regulator; Apoptotic; Behavior; Biochemical; Biological Assay; Biological Factors; Cause of Death; Cell Culture Techniques; Cell Cycle Kinetics; Cell Death; Cell Line; Cells; Cessation of life; Complement; Cultured Cells; Cyclin B; Cytokinesis; Data; Death Rate; Drug resistance; Drug-sensitive; Event; Future; Genotype; Goals; Human; Image; Interphase; Kinetics; Label; Measures; Methods; Microscopy; Mitosis; Mitotic; Mitotic spindle; Molecular; Mus; Mutation; Normal Cell; Outcome; Outer Mitochondrial Membrane; Paclitaxel; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Population; Probability; Proteins; Proteolysis; RNA Interference; Regulation; Reporter; Resistance; S-Phase Fraction; Screening procedure; Testing; Translating; Work; Xenograft procedure; base; cancer cell; cancer therapy; cell killing; chemotherapy; design; drug mechanism; interest; killings; neoplastic cell; novel; prevent; response; small molecule; small molecule libraries; tumor

Our overall goal is to understand the mechanisms by which dividing cancer and normal cells are killed, or escape death, when treated with diverse anti-mitotic drugs. We seek understanding both at the level of cell population behaviors, and the molecular mechanisms that give rise to those behaviors. We are especially interested in understanding how different classes of anti-mitotic drug differ in their ability to kill cancer cells, and using that information to design better future drugs. We will address these goals in 4 specific aims: Aim 1. How do drugs with different anti-mitotic mechanisms differ in their ability to kill cancer cells at the level of cell population behavior? We will use microscopy with fluorescent reporters for key molecular events to measure single-cell responses to four anti-mitotic drugs across a panel of cell lines chosen to vary in apoptosis sensitivity. We will also test the effects of blocking different aspects ofthe drug response. Aim 2. Elucidate the molecular mechanism of cell death during mitotic arrest. To determine how the intrinsic apoptosis pathway is activated by prolonged mitotic arrest we will test candidate regulators of apoptosis, and pursue an unbiased biochemical approach. We will also Identify an alternative cell death pathway cells use when they cannot escape mitotic arrest, and apoptosis is blocked. Aim 3. Develop small molecule inhibitors of mitotic exit that work independent of the SAC. Our preliminary data suggest that novel anti-mitotic drugs with this mechanism would kill cancer cells more effectively than current drugs. We will test this concept, and identify druggable targets in the mitotic exit pathway, by small molecule screening in Core B using a cell-based assay, followed by identification of protein targets of hits. Aim 4. Compare drug responses in mouse tumors to those seen in cell culture. Drug response mechanisms elucidated in cell culture in aims 1-3 may not translate to real tumors. We will test whether cells that die during mitotic arrest in mouse tumors cause bystander killing of non-dividing tumor cells, which could explain why some tumors with low mitotic index can be treated with anti-mitotic drugs. Working with Core C we will probe responses to anti-mitotic drugs in mouse tumors at the single-cell level by intravital imaging.

我们的总体目标是了解杀死癌症和正常细胞的机制，或 逃脱死亡，当用多种抗溶毒药物治疗时。我们寻求在细胞级别上的理解 人口行为以及引起这些行为的分子机制。我们尤其是 有兴趣了解不同类别的抗溶毒药物如何杀死癌细胞的能力不同， 并使用这些信息来设计更好的未来药物。我们将以4个特定目标解决这些目标： AIM 1。具有不同抗隔离机制的药物在杀死癌细胞水平的能力上有何不同 细胞种群行为？我们将使用显微镜与荧光记者一起进行关键分子事件 测量对四种用于变化的细胞系中对四种抗隔离药物的单细胞反应 凋亡敏感性。我们还将测试阻止药物反应不同方面的影响。 目标2。阐明有丝分裂停滞期间细胞死亡的分子机制。确定固有的方式 凋亡途径被延长有丝分裂停滞激活，我们将测试凋亡的候选调节剂，并 采用公正的生化方法。我们还将确定使用替代细胞死亡途径细胞使用 当它们无法逃脱有丝分裂停滞，凋亡被阻塞。 目标3。发展独立于囊的有丝分裂出口的小分子抑制剂。我们的初步 数据表明，具有这种机制的新型抗溶毒药物比 目前的药物。我们将测试这个概念，并通过小 使用基于细胞的测定法，在Core B中筛选分子筛选，然后鉴定出命中的蛋白质靶标。 AIM 4。将小鼠肿瘤中的药物反应与细胞培养中看到的药物反应进行比较。药物反应机制 在AIM 1-3中阐明细胞培养物可能不会转化为实际肿瘤。我们将测试死亡的细胞是否 在小鼠肿瘤中有丝分裂停滞期间导致旁观者杀死非分裂的肿瘤细胞，这可以解释 为什么某些有丝分裂指数低的肿瘤可以用抗丝分裂药物治疗。与Core C合作我们将 探针对小鼠肿瘤对单细胞水平的抗丝分裂药物的反应通过插入式成像。