Dynamics of cellular senescence in single human cells

项目总结/摘要 人类单个细胞衰老的动力学 这项计划的目标是研究单个人类细胞衰老的时间和控制。 衰老是一种永久性细胞周期停滞的状态，是对抗肿瘤进展的内在防御。 利用衰老获得治疗收益将需要更好地理解基本衰老生物学， 新的实验工具，研究在单细胞分辨率的表型，和计算方法， 了解细胞如何整合多种衰老信号。本提案提供了职业过渡计划， 博士这将使他获得在单细胞水平上研究衰老所需的额外训练。 解决并形成开发诱导细胞过早衰老的治疗方法的基础， 有缺陷的压力反应在该奖项的指导阶段（K99），他将开发一个活细胞成像 用于真实的实时监测DNA损伤诱导的衰老的系统以及使用该系统来确定如何 肿瘤抑制因子p53的上游信号传导控制衰老标志物的表达（Aim 1）。这 培训的关键阶段将由Galit Lahav博士和Peter Sorger博士（哈佛医学院）共同监督 他们都是延时显微镜和细胞命运决定过程的专家。在过渡 到独立（K99/R 00），他将确定启动关键基因表达的转录调节因子， 衰老调节因子p16 INK 4a，并表征这种转变的动力学（目的2）。的最后一步 实现独立（R 00）将涉及转移这些工具和概念，以研究衰老， 原代细胞系，特别关注如何整合多种信号以实现衰老 （目标3）。这三个阶段的过渡计划将阐明我们对细胞的基本理解 并提供可扩展的计算/实验平台，用于鉴定 诱导癌细胞过早衰老。这些目标与NIGMS的主要目标高度一致 支持促进人类疾病治疗和预防的基本发现。

Project Summary/Abstract Dynamics of cellular senescence in single human cells The goal of this proposal is to investigate the timing and control of cellular senescence in single human cells. Senescence is a state of permanent cell cycle arrest and an inherent defense against tumor progression. Exploiting senescence for therapeutic gain will require a better understanding of basic senescence biology, novel experimental tools to study the phenotype at single-cell resolution, and computational approaches to understand how cells integrate multiple senescence signals. This proposal provides a career transition plan for Dr. Jeremy Purvis that will equip him with the additional training necessary to study senescence at single-cell resolution and form the foundation for developing treatments that induce premature senescence in cells with defective stress responses. During the mentored phase of the award (K99), he will develop a live-cell imaging system for monitoring DNA damage-induced senescence in real time and use this system to determine how upstream signaling of the tumor suppressor p53 control expression of senescence markers (Aim 1). This critical phase of training will be co-supervised by Dr. Galit Lahav and Dr. Peter Sorger (Harvard Medical School), who are both experts in time-lapse microscopy and cell fate decision processes. During the transition to independence (K99/R00), he will identify transcriptional regulators that initiate expression of the key senescence regulator p16INK4a and characterize the dynamics of this transition (Aim 2). The final step in achieving independence (R00) will involve transferring these tools and concepts to study senescence in primary cells lines, focusing specifically on how multiple signals are integrated to achieve a senescence decision (Aim 3). This three-phase transition plan will illuminate our basic understanding of cellular senescence and provide an extensible computational/experimental platform for identifying therapies that induce premature senescence in cancer cells. These aims are highly congruent with the NIGMS's primary goal of supporting basic discoveries that advance the treatment and prevention of human disease.