ROLE OF CDK2 CELL CYCLE SIGNALING IN ISCHEMIC INJURY AND PROTECTION

CDK2 细胞周期信号转导在缺血性损伤和保护中的作用

• 批准号: 7803551
• 负责人: William Robb MacLellan
• 金额: $ 41.79万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 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/7803551
• 关键词: Adenoviruses; Apoptosis; Apoptotic; Biology; Biophysics; CDK2 gene; Cardiac; Cardiac Myocytes; Caspase; Cell Cycle; Cell Cycle Progression; Cell Cycle Proteins; Cell Cycle Regulation; Cell Death; Cells; Cessation of life; Collaborations; Cyclin A; Data; E2F1 gene; Employee Strikes; Family; Family member; Gene Transfer Techniques; Genes; Genetic; Growth; Heart; Hypoxia; Image Analysis; Infarction; Injury; Investigation; Ischemia; Knockout Mice; MAP Kinase Gene; MAPK14 gene; Mediating; Mitochondria; Mitotic; Modeling; Molecular; Mus; Muscle Cells; Myocardial; Myocardial Ischemia; Myocardium; Nitric Oxide Donors; Pathway interactions; Permeability; Phase; Phosphorylation; Phosphotransferases; Physiology; Play; Principal Investigator; Process; Proteins; Proteomics; Regulation; Relative (related person); Reperfusion Injury; Reperfusion Therapy; Research Personnel; Resistance; Retinoblastoma Genes; Retinoblastoma Protein; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; TP53 gene; Up-Regulation; base; caspase-3; genetic regulatory protein; human CDK2 protein; in vivo; inhibitor/antagonist; interdisciplinary approach; mouse model; novel; numb protein; overexpression; preconditioning; prevent; programs; response; transcription factor

Principal Investigator/ProgramDirector (Last, First, Middle): Ping, Peipei (MacLellan,Project 4) The theme of this Program Project application is to understand the signal transduction pathways mediating ischemic injury and cardioprotection using a multidisciplinary approach combining biophysics, physiology, proteomics and genetics. While Projects 1-3 are focused on two signaling pathways previously known to modulate ischemic injury and protection (i.e., PKCe and p38 MAPK), Project 4 focuses on a pathway with recently recognized importance in the field of myocardial ischemiacell cycle regulatory proteins, in particular, it will focus on three molecules whose functions are interrelated in this pathway: the cyclin-dependent kinase-2 (Cdk2), the retinoblastoma gene product (Rb), and the transcription factors (E2Fs) that regulate genes responsible for cell cycle entry including Cyclin A and E, the catalytic partners of Cdk2. Studies proposed in Project 4 are supported 1) by recent evidence demonstrating that ischemic injury to the heart are accompanied by the upregulation of a number of cell cycle regulatory proteins including Cdk2; and 2) by striking preliminary data demonstrating that infarct size was increased in Rb-null myocardium subjected to regional ischemic injury. Conversely, treatment with a Cdk2 inhibitor in vivo blocked the expected increase in Cdk2 activity with ischemic injury and led to a reduced infarct size formation. This suggests a critical role for Cdk2 in ischemic injury and a cardioprotective role of the Rb protein against injury. Although these observations are intriguing, as the cellular mechanisms underlying these effects are unknown. In collaboration with Projects 1-3 and the Cores, Project 4 proposes 3 Aims. Aim 1 will elucidate the mechanisms underlying Cdk2's ability to modulate ischemic injury; In collaboration with the Heart Biology Core, this aim will determine how Cdk2 modulates mitochondrial function and the cell death pathways; In collaboration with the Proteomic Core, Aim 1 will identify novel Cdk2 substrates. Aim 2 will examine the molecular basis forRb's cardioprotective role in ischemic injury, it will determine p38a MAPK dependent modulation of Rb, and will define the role of E2F family (theprimary targets of Rb) in these processes. Finally, Aim 3 will elucidate mechanisms by which PKCe regulates Cdk2 activity in cardioprotection. These studies will be performed in collaboration with Project 2 and the Heart Biology Core, and will employ two well-established murine models of cardioprotection: the PKCe transgenesis and the nitric oxide donor induced late phase of preconditioning. The proposed investigations will make key contributions to understanding the molecular basis for Rb's cardioprotective effect and the mechanism underlying Cdk2's ability to regulate ischemic damage and apoptotic cell death.

首席研究员/程序导演（最后，第一，中间）：ping，peipei（麦克莱伦，项目4） 该计划项目应用程序的主题是了解介导的信号转导途径 使用多学科方法结合生物物理学，生理学， 蛋白质组学和遗传学。虽然项目1-3集中在以前已知调制的两个信号通路上 缺血性损伤和保护（即PKCE和P38 MAPK），项目4专注于最近认可的途径 在心肌缺血细胞周期调节蛋白方面的重要性，特别是它将集中于三个 在此途径中函数相互关联的分子：细胞周期蛋白依赖性激酶-2（CDK2）， 视网膜母细胞瘤基因产物（RB）和调节负责细胞周期基因的转录因子（E2F） 包括Cyclin A和E的输入，CDK2的催化伴侣。 项目4中提出的研究得到支持1）最近的证据表明，缺血性损伤 心脏伴随着许多细胞周期调节蛋白（包括CDK2）的上调；和2） 引人注目的初步数据表明，在遭受区域性的RB无孔心肌中，梗塞大小增加了 缺血性损伤。相反，用CDK2抑制剂在体内治疗可以阻止CDK2活性的预期增加 缺血性损伤，导致梗塞大小的形成减少。这表明CDK2在缺血性中起关键作用 RB蛋白侵害损伤的损伤和心脏保护作用。尽管这些观察结果很有趣，但 这些作用背后的细胞机制尚不清楚。 与项目1-3和核心合作，项目4提出了3个目标。 AIM 1将阐明 CDK2调节缺血性损伤能力的机制；与心脏生物学核心合作，这 AIM将决定CDK2如何调节线粒体功能和细胞死亡途径；与 蛋白质组学核心，AIM 1将识别新颖的CDK2底物。 AIM 2将检查分子基础FORB的基础 心脏保护作用在缺血性损伤中，它将确定RB的P38A MAPK依赖性调制，并将定义 E2F家族（RB的主要靶标）在这些过程中的作用。最后，AIM 3将通过 PKCE调节心脏保护中的CDK2活性。这些研究将与项目合作进行 2和心脏生物学核心，并将采用两种成熟的鼠类心脏保护模型：PKCE 转带和一氧化氮供体诱导了预处理的后期。拟议的调查将 为理解RB心脏保护作用的分子基础做出关键贡献和机制 CDK2调节缺血性损伤和凋亡细胞死亡的潜在能力。