Novel functions of D-type and E-type cyclins in normal and in cancer cells

D 型和 E 型细胞周期蛋白在正常细胞和癌细胞中的新功能

基本信息

批准号：
9886203
负责人：
Peter Sicinski
金额：
$ 43.1万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9886203
关键词：
Ablation Acute Address Affect Animals Automobile Driving Binding CCNE1 gene CDK2 gene CDK4 gene Cell Cycle Cell Cycle Progression Cell Differentiation process Cell Proliferation Cells Characteristics Complex Cyclin D1 Cyclin-Dependent Kinases Cyclins Down-Regulation G1 Phase Glioblastoma Grant Growth Human Immunocompromised Host In Vitro Injections Knockout Mice Laboratories Literature Maintenance Malignant - descriptor Malignant Neoplasms Malignant neoplasm of brain Mammalian Cell Molecular Molecular Analysis Mus Nervous system structure Neurons Normal Cell Oncogenic Pathway interactions Patients Phenotype Phosphotransferases Play Proliferating Property Proteins Research Role Set protein Substrate Specificity System Teratoma Testing Therapeutic Time Undifferentiated Work blastocyst cancer cell cancer cell differentiation cancer type cell type conditional knockout cyclin G1 design embryonic stem cell in vivo knock-down mouse model neurogenesis novel novel therapeutics overexpression pluripotency programs protein function public health relevance stem cells stem-like cell stemness subcutaneous tool treatment strategy tumor tumorigenesis tumorigenic

项目摘要

DESCRIPTION (provided by applicant): The proliferation of mammalian cells is driven by the core cell cycle machinery. The key components of this machinery are proteins called cyclins, which bind, activate and provide substrate specificity to the cyclin- dependent kinases (CDKs). Cyclin-CDK complexes phosphorylate cellular proteins, thereby driving cell proliferation. Cyclins operating during the G1 phase of the cell cycle (the so-called "G1 cyclins") are of particular importance to the cancer field, as many oncogenic pathways were shown to impinge on these proteins. Two classes of G1 cyclins operate in mammalian cells: D-type (cyclins D1, D2 and D3) and E-type (E1 and E2). D-cyclins activate CDK4 and CDK6, whereas E-cyclins activate mostly CDK2. Cyclin D-CDK4, D-CDK6 and E-CDK2 complexes phosphorylate an overlapping set of proteins; cyclin E-associated kinase is thought to target a broader spectrum of substrates. In the past, our laboratory generated and characterized mice lacking D-type or E-type cyclins. Although these animals displayed focused abnormalities, the majority of cell types proliferated normally or nearly normally in the absence of D-type or E-type cyclins. We ascribed the absence of more profound phenotypes to the overlapping functions of the two G1 cyclin classes. To better understand the in vivo functions of G1 cyclins, we generated conditional quintuple knockout mouse embryonic stem cells. These cells allow us to study the consequences of an acute shutdown of all five G1 cyclins in embryonic stem cells. In the proposed work, we will use this experimental system to study the molecular functions of G1 cyclins. An important part of our research plan are analyses of G1 cyclin functions in mouse and in human cancer cells. We will focus on glioblastoma, the most aggressive and incurable brain cancer. We will investigate the molecular functions of G1 cyclins in this tumor type, using a mouse model of glioblastoma as well as primary patient-derived glioblastoma cells. We will test whether inhibition of G1 cyclin function might represent a therapeutic strategy in this deadly cancer type. We will address the following major issues in our three Specific Aims: In Specific Aim 1, we will study the function of G1 cyclins in embryonic stem cells. In Specific Aim 2, we will investigate the molecular functions of G1 cyclins during differentiation. Lastly, in Specific Aim 3, we will extend our studies to mous and human patient-derived cancer cells, by analyzing the function of G1 cyclins in glioblastoma cells. Our work may suggest novel therapeutic strategies for treatment of glioblastoma. This tumor represents the most aggressive and incurable brain cancer, and the median survival time of the affected patients is currently less than one year.

描述（由申请人提供）：哺乳动物细胞的增殖是由核心细胞周期机制驱动的，该机制的关键成分是称为细胞周期蛋白的蛋白质，其结合、激活细胞周期蛋白依赖性激酶（CDK）并为其提供底物特异性。细胞周期蛋白-CDK 复合物磷酸化细胞蛋白，从而驱动细胞增殖，在细胞周期的 G1 期运行的细胞周期蛋白（所谓的“G1 细胞周期蛋白”）尤其重要。哺乳动物细胞中有两类 G1 细胞周期蛋白：D 型（细胞周期蛋白 D1、D2 和 D3）和 E 型（E1 和 E2）。细胞周期蛋白激活 CDK4 和 CDK6，而 E 细胞周期蛋白主要激活 CDK2、细胞周期蛋白 D-CDK4、D-CDK6 和 E-CDK2 复合物。重叠的蛋白质组；细胞周期蛋白 E 相关激酶被认为针对广泛的底物在过去，我们的实验室生成并表征了缺乏 D 型或 E 型细胞周期蛋白的小鼠，尽管这些动物表现出集中的异常。在缺乏 D 型或 E 型细胞周期蛋白的情况下，细胞类型正常或几乎正常增殖，我们将更深刻的表型的缺乏归因于两个 G1 细胞周期蛋白类别的重叠功能。 G1 细胞周期蛋白，我们产生了条件性五重敲除小鼠胚胎干细胞，这些细胞使我们能够研究胚胎干细胞中所有五种 G1 细胞周期蛋白急性关闭的后果。在拟议的工作中，我们将使用该实验系统来研究分子。 G1 细胞周期蛋白的功能我们研究计划的一个重要部分是分析小鼠和人类癌细胞中的 G1 细胞周期蛋白功能我们将重点关注胶质母细胞瘤，这是最具侵袭性且无法治愈的大脑。我们将使用胶质母细胞瘤小鼠模型以及原发性患者来源的胶质母细胞瘤细胞来研究 G1 细胞周期蛋白在这种肿瘤类型中的分子功能。我们将测试 G1 细胞周期蛋白功能的抑制是否可能代表这种致命癌症的治疗策略。我们将在三个具体目标中解决以下主要问题：在具体目标 1 中，我们将研究的功能。胚胎干细胞中的 G1 细胞周期蛋白。在特定目标 2 中，我们将研究分化过程中 G1 细胞周期蛋白的分子功能。最后，在特定目标 3 中，我们将通过分析功能将研究扩展到小鼠和人类患者来源的癌细胞。我们的工作可能为治疗胶质母细胞瘤提供新的治疗策略，这种肿瘤代表了最具侵袭性和无法治愈的脑癌，以及受影响患者的中位生存时间。目前还不到一年的时间。