Defining the role of ERK1 and ERK2 in Pancreatic Cancer

定义 ERK1 和 ERK2 在胰腺癌中的作用

• 批准号: 8835707
• 负责人: Meagan B Ryan
• 金额: $ 3.35万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-04 至 2018-02-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8835707
• 关键词: Accounting; Adenocarcinoma Cell; Adhesions; Agar; Anchorage-Independent Growth; Apoptosis; Automobile Driving; BRAF gene; Biological Assay; Biological Process; Cancer Biology; Cancer Etiology; Cell Cycle Arrest; Cell Cycle Regulation; Cell Line; Cell physiology; Cells; Cessation of life; Dependency; Development; Disease; Docking; Doxycycline; E-Cadherin; Ectopic Expression; Embryo; Employee Strikes; Epithelial; Event; Fellowship; Flow Cytometry; Foundations; Genes; Genetic; Genetically Engineered Mouse; Glycolysis; Goals; Growth; Immunofluorescence Immunologic; KRAS2 gene; Knockout Mice; Lead; Left; Literature; MAPK1 gene; MAPK3 gene; MEKs; Maintenance; Malignant Neoplasms; Malignant neoplasm of pancreas; Malignant neoplasm of prostate; Mass Spectrum Analysis; Mediating; Melanoma Cell; Mesenchymal; Metabolic; Migration Assay; Mitogen-Activated Protein Kinases; Mutate; Pancreas; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Peptides; Phosphorylation; Phosphotransferases; Play; Ploidies; Process; Propidium Diiodide; Protein Isoforms; Protein Kinase; Proteins; Ras/Raf; Reporting; Research; Research Personnel; Research Technics; Role; STAT1 gene; Sampling; Series; Signal Transduction; Site; Staining method; Stains; Survival Rate; Testing; Therapeutic Intervention; Tumor Cell Invasion; United States; Vimentin; Western Blotting; annexin A5; anticancer research; base; c-myc Genes; cell behavior; cell motility; epithelial to mesenchymal transition; genetic regulatory protein; knock-down; knowledge base; malignant breast neoplasm; migration; mouse model; mutant; novel; outcome forecast; public health relevance; research study; senescence; skills; small hairpin RNA; standard care; therapeutic target; transcription factor; tumor

 DESCRIPTION (provided by applicant): Pancreatic cancer is the fourth leading cause of cancer deaths in the United States, with a poor prognosis and limited treatment options. Pancreatic ductal adenocarcinoma (PDAC) accounts for the greatest number of these cases. The gene KRAS is mutated in greater than 95% of PDAC cases, and this leads to the activation of the Raf-MEK-ERK mitogen-activated protein kinase (MAPK) pathway. Substantial experimental evidence implicates the key role of this protein kinase cascade in driving KRAS-dependent PDAC growth. Despite this importance, the mechanisms by which ERK drives PDAC growth remain surprisingly unresolved. At every level of the Ras-Raf-MEK-ERK cascade there are structurally and functionally related isoforms. There is now strong evidence for striking differences in the role of different Ras and Raf isoforms in signaling and cancer. Whether this is also the case for the two highly related ERK1 and ERK2 isoforms, which share 84% sequence identity, remains underexplored. Much of our current understanding of ERK biological function comes from studies on "ERK1/2" that did not distinguish between these two isoforms. In addition, ERK1/2 are capable of phosphorylating greater than 200 known substrates, but there is little understanding of which of these substrates are important drivers of the ERK/MAPK pathway. The extensive list of substrates causes further complexity for studies regarding which substrates play an active role in the development and maintenance of pancreatic cancer. Based on some reports in the literature suggesting that ERK1 and ERK2 may play different roles in certain circumstances, I performed preliminary studies to determine whether these two isoforms are interchangeable in PDAC. My studies using shRNA directed against ERK1 or ERK2 in a panel of 10 PDAC cell lines showed that loss of ERK1 or ERK2 reduced both anchorage-dependent clonogenic growth and anchorage-independent soft agar growth, indicating that neither of these highly related isoforms can compensate functionally for the loss of the other. Therefore, I hypothesize that ERK1 and ERK2 have distinct and nonredundant roles in driving KRAS-mutant pancreatic cancer growth. I propose three specific aims to test my hypothesis: I will (1) define the dependency of KRAS-mutant PDAC growth on ERK1 and ERK2; (2) define ERK1 and ERK2 roles in PDAC epithelial to mesenchymal transformation, invasion and metabolic alterations; and, (3) Identify ERK isoform-dependent substrates in PDAC. These studies will investigate the divergent and shared functions of ERK1 and ERK2 in pancreatic cancer and have the potential to discover novel signaling mechanisms of this critical node in the ERK MAPK pathway. This fellowship will also enhance my research skills, establish a firm foundation in basic and translational cancer biology and help develop my skills to become an independent investigator in cancer research.

 描述（由适用提供）：胰腺癌是美国癌症死亡的第四个主要原因，预后较差，治疗方案有限。胰腺导管腺癌（PDAC）占此类病例中最多的数量。在大于95％的PDAC病例中突变基因KRAS，这导致RAF-MEK-ERK促分裂原激活的蛋白激酶（MAPK）途径的激活。大量的实验证据实现了该蛋白激酶级联反应在驱动KRAS依赖性PDAC生长中的关键作用。尽管这一点很重要，但ERK驱动PDAC增长的机制仍然令人惊讶地尚未解决。在RAS-RAF-MEK-ERK级联的每个级别上都有结构和功能相关的同工型。现在有强有力的证据表明，不同RAS和RAF同工型在信号传导和癌症中的作用上存在差异。对于具有84％序列身份的两个高度相关的ERK1和ERK2同工型也是如此。我们目前对ERK生物学功能的大部分理解都来自对“ ERK1/2”的研究，这些研究没有区分这两种同工型。此外，ERK1/2能够磷酸化大于200个已知底物，但是对这些底物中的哪些是ERK/MAPK途径的重要驱动因素几乎没有什么了解。关于哪些底物在胰腺癌的发展和维持中起着积极作用的研究，广泛的底物列表引起了进一步的复杂性。根据文献中的一些报告，表明ERK1和ERK2在某些情况下可能扮演着不同的角色，我进行了初步研究，以确定这两种同工型在PDAC中是否可以互换。我使用针对ERK1或ERK2的SHRNA的研究在一组10个PDAC细胞系中的研究表明，ERK1或ERK2的损失降低了锚固依赖性的结合生长和与锚固无关的软琼脂的生长，这表明这些高度相关的同工型都无法在功能上补偿其他人的损失。因此，我假设ERK1和ERK2在推动KRAS突变胰腺癌生长方面具有独特和非冗余的作用。我提出了三个特定的目的来检验我的假设：我将（1）定义KRAS突变PDAC生长对ERK1和ERK2的依赖性； （2）将ERK1和ERK2在PDAC上皮中定义为间质转化，侵袭和代谢改变； （3）识别PDAC中的ERK同工型依赖性底物。这些研究将研究ERK1和ERK2在胰腺癌中的分歧和共享功能，并有可能发现ERK MAPK途径中这种关键节点的新型信号传导机制。这项奖学金还将提高我的研究技能，在基本和翻译的癌症生物学上建立牢固的基础，并帮助发展我成为癌症研究独立研究者的技能。