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（由申请人提供）：胰腺癌是美国第四大癌症死亡原因，其中胰腺导管腺癌 (PDAC) 的病例数量最多，且预后不良且治疗选择有限。超过 95% 的 PDAC 病例发生突变，这会导致 Raf-MEK-ERK 丝裂原激活蛋白激酶 (MAPK) 通路的激活。大量实验证据表明，这种蛋白激酶级联在驱动 KRAS 依赖性 PDAC 生长中发挥着关键作用，尽管如此，ERK 驱动 PDAC 生长的机制在 Ras-Raf-MEK-ERK 级联的各个层面上仍然没有得到解决。是结构和功能相关的异构体，现在有强有力的证据表明不同 Ras 和 Raf 异构体在信号传导和癌症中的作用存在显着差异。两种高度相关的 ERK1 和 ERK2 异构体具有 84% 的序列同一性，目前我们对 ERK 生物学功能的大部分了解仍来自于对“ERK1/2”的研究，这些研究并未区分这两种异构体。 /2 能够磷酸化超过 200 种已知底物，但人们对这些底物中哪些是 ERK/MAPK 途径的重要驱动因素知之甚少。底物的广泛列表导致关于哪些底物在胰腺癌的发展和维持中发挥积极作用的研究变得更加复杂。根据文献中的一些报告表明 ERK1 和 ERK2 在某些情况下可能发挥不同的作用，我进行了初步研究。确定这两种亚型在 PDAC 中是否可以互换。我在一组 10 个 PDAC 细胞系中使用针对 ERK1 或 ERK2 的 shRNA 进行的研究表明，ERK1 或 ERK2 的缺失会降低两者的水平。贴壁依赖性克隆生长和贴壁非依赖性软琼脂生长，表明这两种高度相关的亚型都不能在功能上补偿另一种亚型的损失，因此，我着迷于 ERK1 和 ERK2 在驱动 KRAS 突变胰腺中具有独特且非冗余的作用。我提出三个具体目标来检验我的假设： (1) 定义 KRAS 突变 PDAC 生长对 ERK1 和 ERK2 的依赖性； (2) 定义 ERK1 和 ERK2 在 PDAC 上皮间质转化、侵袭和代谢改变中的作用；(3) 鉴定 PDAC 中 ERK 异构体依赖性底物。这些研究将研究 ERK1 和 ERK2 在胰腺癌中的不同和共同功能。并有潜力发现 ERK MAPK 通路中这个关键节点的新信号传导机制。这项奖学金也将提高我的研究技能，为基础和研究奠定坚实的基础。转化癌症生物学并帮助我发展技能，成为癌症研究的独立研究者。