Inhibition of pancreatic carcinogenesis via targeting c-Raf and sEH

通过靶向 c-Raf 和 sEH 抑制胰腺癌发生

• 批准号: 9056497
• 负责人: Guang-Yu Yang
• 金额: $ 32.03万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9056497
• 关键词: Acids; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Arachidonic Acids; BAY 54-9085; Benzoic Acids; Blood; Caerulein; Cancer cell line; Cells; Cytochrome P450; Data; Development; Dose; Drug Kinetics; Early Diagnosis; Ensure; Enzymes; Epoxide hydrolase; Event; Exhibits; Gene Expression; Gene Mutation; Growth; Health; Human; Hydrolase; In Vitro; Inflammation; Inflammatory; Knock-out; Lead; MAP Kinase Gene; MEKs; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mediator of activation protein; Modeling; Molecular; Mus; Mutation; NF-kappa B; Oral; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Ductal Carcinoma; Pancreatic Intraepithelial Neoplasia; Pancreatitis; Phosphorylation; Picolinic Acids; Play; Prevention; Prevention strategy; Risk Factors; Role; Sampling; Signal Transduction; TP53 gene; Testing; Time; Toxic effect; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Growth Factors; attenuation; base; carcinogenesis; chronic pancreatitis; cytokine; design; flexibility; in vivo; inhibitor/antagonist; kinase inhibitor; knockout gene; mortality; mouse model; mutant; novel; overexpression; pancreatic tumorigenesis; small molecule; success; tumor; tumorigenesis

DESCRIPTION (provided by applicant): The significance of this project is to develop an efficient agent for the prevention of highly lethal pancreatic cancer through targeting both c-Raf and sEH. Early detection and prevention is of great importance to reduce the mortality of pancreatic cancer. Mutant Kras is the most common and earliest molecular event involved in human pancreatic carcinogenesis; and using mutant Kras-initiated tumorigenesis in mice, it has been demonstrated that c-Raf is required for Kras-initiated tumorigenesis and knockout c-Raf blocks Kras- initiated cancer development. Chronic pancreatitis is a well-recognized risk factor of pancreatic cancer. In mutant Kras-initiated pancreatic tumorigenesis in Pdx1-Cre/LSL-KrasG12D mice (called PanKras), caerulein- induced pancreatitis enhances malignant progression. In addition to pancreatitis, mutant Kras cooperates with mutant p53 (the second most common genetic alteration) leading to pancreatic cancer development in mice, mainly via synergistically regulating ERK-MAPK and NF-kB. Anti-inflammatory EETs are quickly inactivated by pro-inflammatory soluble epoxide hydrolase (sEH). Our preliminary data showed that 1) 96% (27/28) and 90% (27/30) of human pancreatic adenocarcinoma samples displayed a distinct over-expression of sEH and p-c-Raf, respectively, and 2) our novel dual c-Raf/sEH inhibitor t-CUPM exhibited strong inhibitory effect on pancreatitis and mPanIN formation in PanKras mice. Our hypothesis is that c-Raf and sEH play key roles in pancreatic carcinogenesis initiated by mutant Kras and enhanced by pancreatitis or mutant p53; and targeting these two enzymes together is a novel and relevant preventive strategy for pancreatic cancer. Three specific aims are proposed to test this hypothesis: 1) to determine the effects of c-Raf/sEH dual inhibitor t-CUPM, sEH inhibitor t- AUCB and Raf inhibitor Raf265/sorafenib on mutant Kras-initiated and pancreatitis-enhanced carcinogenesis in PanKras mice; 2) to determine whether blocking mutant Kras-activated c-Raf using inhibitors or gene knockout suppresses mutant Kras-p53 interaction and malignant progression in Pdx1-KrasG12D-p53R172 mice; and 3) to determine the role of sEH in pancreatitis and carcinogenesis using the approach of sEH gene knockout in PanKras mice. Successfulness of the proposed study will not only lead us to demonstrate the role of c-Raf and sEH in pancreatic carcinogenesis, but also to establish a significance of t-CUPM as a novel agent for inhibiting the pancreatic cancer development.

描述（由申请人提供）：该项目的意义在于开发一种通过靶向c-Raf和sEH来预防高致死性胰腺癌的有效药物。早期发现和预防对于降低胰腺癌的死亡率具有重要意义。 Kras突变是人类胰腺癌发生过程中最常见、最早的分子事件；通过在小鼠中使用突变型 Kras 引发的肿瘤发生，已经证明 c-Raf 是 Kras 引发的肿瘤发生所必需的，并且敲除 c-Raf 可以阻止 Kras 引发的癌症发展。慢性胰腺炎是公认的胰腺癌危险因素。在 Pdx1-Cre/LSL-KrasG12D 小鼠（称为 PanKras）中突变的 Kras 引发的胰腺肿瘤发生中，雨蛙素诱导的胰腺炎增强了恶性进展。除了胰腺炎之外，突变型 Kras 与突变型 p53（第二常见的基因改变）协同作用，主要通过协同调节 ERK-MAPK 和 NF-kB，导致小鼠胰腺癌的发生。抗炎 EET 很快就会被促炎可溶性环氧化物水解酶 (sEH) 灭活。我们的初步数据显示：1) 96% (27/28) 和 90% (27/30) 的人胰腺腺癌样本分别表现出 sEH 和 p-c-Raf 的明显过度表达，2) 我们的新型 c-Raf/sEH 双抑制剂 t-CUPM 对 PanKras 小鼠的胰腺炎和 mPanIN 形成表现出强烈的抑制作用。我们的假设是，c-Raf 和 sEH 在由突变 Kras 引发并由胰腺炎或突变 p53 增强的胰腺癌发生中发挥关键作用；同时针对这两种酶是一种新颖且相关的胰腺癌预防策略。提出了三个具体目标来检验这一假设：1）确定c-Raf/sEH双重抑制剂t-CUPM、sEH抑制剂t-AUCB和Raf抑制剂Raf265/索拉非尼对PanKras小鼠中突变型Kras引发和胰腺炎增强的致癌作用的影响； 2) 确定使用抑制剂或基因敲除阻断突变体 Kras 激活的 c-Raf 是否会抑制 Pdx1-KrasG12D-p53R172 小鼠中突变体 Kras-p53 相互作用和恶性进展； 3) 利用 PanKras 小鼠中 sEH 基因敲除的方法确定 sEH 在胰腺炎和癌症发生中的作用。这项研究的成功不仅使我们证明了 c-Raf 和 sEH 在胰腺癌发生中的作用，而且还确立了 t-CUPM 作为抑制胰腺癌发展的新型药物的重要性。