Parameters that determine cell fate during mitotic arrest

有丝分裂停滞期间决定细胞命运的参数

基本信息

批准号：
10617385
负责人：
Charles Bradley Shuster
金额：
$ 14.4万
依托单位：
NEW MEXICO STATE UNIVERSITY LAS CRUCES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-10 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10617385
关键词：
4D Imaging Acceleration Adjuvant Therapy Antimitotic Agents Apoptosis Apoptotic BCL2 gene Biochemical Biosensor Cancer cell line Cell Cycle Cell Death Cell Line Cell Survival Cell division Cells Centrioles Chromosome Segregation Clinic Cytoprotection Data Dependence Exhibits Failure Family Flow Cytometry Foundations Future Goals Growth Hela Cells Heterogeneity Hormones Image Individual Jordan Kinesin Kinetics Laboratories MCL1 gene Malignant Neoplasms Metabolism Microscopy Microtubules Mitosis Mitotic Mitotic spindle Molecular Molecular Motors Normal Cell Pathway interactions Peripheral Nervous System Diseases Pharmaceutical Preparations Phosphotransferases Proliferating Protein Inhibition Proteins Reagent Regulation Regulatory Pathway Resolution Role Signal Pathway Signal Transduction Structure TP53 gene Therapeutic Vinblastine Vinca Alkaloids Western Blotting Work cancer cell cancer therapy cell behavior chemotherapy drug resistance development graduate student imaging approach inhibitor insight live cell imaging mTOR Signaling Pathway member neoplastic cell population based response side effect taxane training opportunity translational study tumor undergraduate student

项目摘要

Prolonged activation of the spindle assembly checkpoint (SAC) due to mitotic spindle disruption can result in p53 activation, centriole disengagement and cell death. Indeed, one chemotherapeutic strategy frequently applied to aggressive and hormone-independent cancers is to target the mitotic spindle, a microtubule-based structure that is required for proper chromosome segregation and cell division. Drugs such as vinblastine or Paciltaxel suppress the normal microtubule assembly dynamics, leading to mitotic arrest and eventual cell death by apoptosis. However, despite their decades-long implementation in the clinic, the mechanisms by which prolonged mitotic delay results in cell death remains unclear. Further, despite the universality of the requirement of the mitotic spindle for cell division, there is still a great deal of heterogeneity in how cells respond to spindle disruption, which may reduce the efficacy of anti- mitotic chemotherapeutic strategies. Using a combination of biochemical and live cell imaging approaches, our preliminary data reveals that targeting both Kinesin Spindle Protein (KSP), a molecular motor required for spindle bipolarity, and the Phosphatidylinositide 3-kinase (PI3K)/Akt/mTOR signaling pathway dramatically accelerates the kinetics of mitotic cell death relative to mitotic arrest alone. Moreover, it elicits a more homogeneous response from the treated cells. PI3K signaling is involved in a variety of regulatory pathways that regulate cell survival, metabolism and proliferation, but the mechanism by which PI3K activity promotes cell viability during mitotic arrest is unknown. To better understand how PI3K signaling is involved in the timing of cell death and variability of cellular responses of mitotic delay, we will continue to apply high-throughput timelapse imaging, high-resolution 4D imaging and biochemical approaches to a battery of cell lines differ in their sensitivity to mitotic delay as well as dependence on PI3K signaling. The Specific Aims of this project will 1) Define the protective role of PI3K in normal and cancer cells; and 2) Determine the mechanism by which PI3K promotes cell survival during mitotic delay. If successful, these studies will lay the foundation for future translational studies to further develop adjuvant therapies that will target mitotically active tumor cells without the side effects associated with other microtubule disruptors.

由于有丝分裂纺锤体破坏导致纺锤体装配检查点 (SAC) 延长激活可导致 p53 激活、中心粒脱离和细胞死亡。确实，一化疗策略经常应用于侵袭性和激素非依赖性癌症是针对有丝分裂纺锤体，这是一种基于微管的结构，是正常细胞分裂所必需的染色体分离和细胞分裂。长春花碱或紫杉醇等药物可抑制正常的微管组装动力学，导致有丝分裂停滞并最终细胞死亡细胞凋亡。然而，尽管这些机制在临床上实施了数十年，但长期有丝分裂延迟导致细胞死亡尚不清楚。此外，尽管尽管细胞分裂对有丝分裂纺锤体的要求具有普遍性，但仍然存在大量的问题。细胞如何应对纺锤体破坏的异质性，这可能会降低抗- 有丝分裂化疗策略。结合使用生化和活细胞成像方法，我们的初步数据表明，针对驱动蛋白纺锤蛋白（KSP），纺锤体双极所需的分子运动和磷脂酰肌醇 3-激酶 (PI3K)/Akt/mTOR 信号通路显着加速有丝分裂细胞死亡动力学相对于单独的有丝分裂停滞。此外，它还引起了更一致的反应处理过的细胞。 PI3K信号传导参与调节细胞的多种调控途径生存、代谢和增殖，但PI3K活性促进细胞的机制有丝分裂停滞期间的活力尚不清楚。为了更好地了解 PI3K 信号如何参与细胞死亡的时间和有丝分裂延迟的细胞反应的可变性，我们将继续应用高通量延时成像、高分辨率4D成像和生化细胞系电池组的方法不同之处在于它们对有丝分裂延迟的敏感性以及对 PI3K 信号传导的依赖。该项目的具体目标将 1) 定义保护措施 PI3K 在正常细胞和癌细胞中的作用； 2) 确定 PI3K 的机制促进有丝分裂延迟期间的细胞存活。如果成功的话，这些研究将为未来的转化研究进一步开发针对有丝分裂活性的辅助疗法肿瘤细胞没有与其他微管破坏剂相关的副作用。