Roles and Regulation of wild-type and mutant forms of p53

总体摘要 在过去的15年里，我们的项目CA87497一直具有很高的生产力和互动性。 该计划的未来目标是研究与上下文相关的肿瘤抑制作用。 野生型P53，并阐明癌症相关突变型P53如何促进肿瘤的发生。 我们将研究新发现的野生型和突变型P53蛋白的途径 以及它们各自在调控表观基因组中的作用和新的特征 P53的转录靶点。除了现任成员，Carol Prives博士，Arnold 莱文、斯科特·洛和卡洛斯·科登-卡多，我们请来了布伦特·斯托克韦尔博士 John Petrini，每个人都贡献了新的和令人兴奋的方向，与现有的 项目。我们的方法包括细胞生物学、化学生物学、生物化学、蛋白质组学、 显微镜、生物信息学、功能基因组学、小鼠建模和人与鼠 病理学。我们的研究具有很高的翻译性，与人类疾病相关，重点是 乳腺癌、肝癌和淋巴瘤。项目1(Prives)将研究突变型p53基因的获得 使用基于细胞的分析、基因表达谱和蛋白质组学的功能活动。 项目1将阐明突变型p53如何以及为什么在野生型的情况下刺激甲羟戊酸途径。 P53类型抑制这一相同的途径，并将获得关于如何 突变型P53促进核小体重塑。项目2(斯托克韦尔)将定义机制 这是他的研究小组发现的，它控制着P53对铁性下垂的调控。计划的工作将定义 P53靶基因p21如何在负反馈环中起作用以抑制铁性下垂 通过对甲氧戊酸途径的影响来调节铁下垂，以及营养缺乏如何 调节P53及其对铁性下垂的影响。项目3(Petrini)将量化并确定 癌基因激活引起的P53依赖性表观遗传学改变在乳腺中的定位 并将确定这一新途径是否在造血细胞中起作用， 并评估其对抑制淋巴增生症和髓系白血病的重要性。项目 4(LOWE)使用先进的遗传和基因组工具以及新的基因工程 用小鼠模型探讨P53在不同组织中抑制肿瘤的机制 和遗传背景，以及新的p53突变体在肿瘤发生和发展中的作用 活着。它还将探索铁性下垂在肿瘤抑制中的作用，以及如何放松对 包括甲氧戊酸途径在内的P53介导的基因抑制程序与肿瘤有关 维修。计划中的研究将广泛依赖三个核心：核心A(Prives)，即 将对生物信息学核心B计划内的所有互动提供行政支持 (Levine)将为每个项目提供关键的计算支持，以帮助 突变型和野生型p53在不同环境中调控基因和途径的发现。 核心B将测试新的假说，并确定野生型和突变型p53的新调节器。这个 分子系统病理学核心C(Cordon-Cardo)将提供以下关键信息 人类肿瘤包括甲氧丙戊酸和铁下垂的新分子生物标记物 小路。核心C还将量化异色标记，并将研究突变的生物标记物 肝和淋巴瘤样本中的P53、干细胞和新的抑制靶点。

Overall Summary For the past 15 years our Program Project CA87497 has been highly productive and interactive. The future goals of this program are to study the context-dependent tumor suppressive roles of wild-type p53, and to elucidate how cancer related mutant forms of p53 promote oncogenesis. We will investigate newly discovered pathways in which wild-type and mutant p53 proteins operate and their respective roles in regulating the epigenome and characterize new transcriptional targets of p53. In addition to the current members, Drs Carol Prives, Arnold Levine, Scott Lowe and Carlos Cordon-Cardo, we have brought in Drs Brent Stockwell and John Petrini who each contribute new and exciting directions that interface with the existing projects. Our approaches include cell biology, chemical biology, biochemistry, proteomics, microscopy, bioinformatics, functional genomics, mouse modeling and human and mouse pathology. Our research is highly translational and relevant to human disease, focusing on breast cancer, liver cancer and lymphoma. Project 1 (Prives) will investigate mutant p53 gain of function activities employing cell-based assays, gene expression profiling and proteomics. Project 1 will elucidate how and why mutant p53 stimulates the mevalonate pathway, while wild- type p53 represses this same pathway, and will obtain mechanistic information as to how mutant p53 facilitates nucleosome remodeling. Project 2 (Stockwell), will define mechanisms that govern p53 regulation of ferroptosis, which his group discovered. Planned work will define how the p53 target p21 acts in a negative feedback loop to restrain ferroptosis, how p53 regulates ferroptosis through its effects on the mevalonate pathway, and how nutrient deficiency regulates p53 and its impact on ferroptosis. Project 3 (Petrini) will quantify and determine the location of p53-dependent epigenetic changes induced by oncogene activation in mammary epithelium and will determine whether this novel pathway is operative in hematopoietic cells, and assess its importance for suppression of lymphomagenesis and myeloid leukemia. Project 4 (Lowe) uses advanced genetic and genomic tools together with novel genetically-engineered mouse models to explore mechanisms of p53-mediated tumor suppression in different tissue and genetic contexts, and the role of novel p53 mutants on tumor initiation and progression in vivo. It will also explore the role of ferroptosis in tumor suppression and how deregulation of p53-mediated gene repression programs including the mevalonate pathway contributes to tumor maintenance. The planned research will rely extensively on three cores: Core A (Prives), that will administratively support all interactions within the program, the Bioinformatics Core B (Levine) that will provide crucial computational support for each project in order to aid in the discovery of genes and pathways regulated by mutant and wild-type p53 in different contexts. Core B will test new hypotheses and identify new modulators of wild-type and mutant p53. The Molecular Systems Pathology Core C (Cordon-Cardo) will provide critical information regarding human tumors including new molecular biomarkers derived from mevalonate and ferroptotic pathways. Core C will also quantify heterochromatic marks and will study biomarkers of mutant p53, stem cells, and new repression targets in liver and lymphoma samples.