ERK PATHWAYS IN PATHOGENESIS OF MESOTHELIOMA

间皮瘤发病机制中的 ERK 通路

• 批准号: 7035622
• 负责人: Brooke Taylor Mossman
• 金额: $ 32.82万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7035622
• 关键词: antineoplastics; asbestos; athymic mouse; carcinogens; carmustine; cell proliferation; cocarcinogen; environment related neoplasm /cancer; enzyme activity; enzyme mechanism; gene environment interaction; gene expression; genetically modified animals; human tissue; mesothelioma; mitogen activated protein kinase; neoplastic transformation; silicates; simian virus 40; virus related neoplasm /cancer; xenotransplantation

Extracellular signal regulated kinases, ERK1/2 and ERK5, are activated by asbestos fibers in mesothelial and airway epithelial cells and play critical roles in cell survival. ERK1/2-dependent Fra-1 expression is also linked causally to morphologic transformation of rat mesothelial cells and expression of genes (c-met, cd44) stimulating cell proliferation and migration. We hypothesize that activation of ERK1/2 and ERK5 signaling pathways occur by carcinogenic fibers (asbestos, erionite) in the pathogenesis of human malignant mesothelioma (MM) and are potentiated by SV40 in a co-carcinogenic manner. Recent exciting data also suggest that these survival pathways are activated in MMs after exposure to chemotherapeutic drugs and can be manipulated to achieve increased cell killing. Thus, we also hypothesize that ERK1/2 and ERK5 pathways contribute, alone or cooperatively, to MM cell survival after chemotherapy. In Aim 1, we will test crocidolite and chrysotile asbestos, well-characterized Turkish erionite (see Project 1), and their nonfibrous analogs, alone and with co-exposures to SV40 to determine if ERK1/2, and ERK5 activity, fos/jun family members, and AP-1 transactivation correlate with patterns of transformation and carcinogenicity as determined in the in vitro/in vivo models developed by Dr. Carbone (Core C). In Aim 2, we will use a panel of SV40+ and - MMs from Core B to determine the effects of dominant negative constructs and small hairpin (sh) RNA interference (RNAi) vectors targeting ERK1/2 and ERK5 on parameters of in vitro cell transformation and survival after treatment with Carmustine (BCNU). In collaboration with Dr. Testa (Project 3) we will also determine if the AKT survival pathway is modified in these studies. In Aim 3, a mouse orthotopic model will be used to determine if SV40+ and - MMs stably transfected with shMEKI, shERKS or both constructs before intrapleural injection and administration of Carmustine, have altered growth and metastases. Identifying the pathways of cell survival in the pathogenesis of MMs is highly significant for prevention and treatment of these devastating tumors. This Program Project allows our previous mechanistic studies in rodent mesothelioma models to be validated in human mesotheliomas and provides us with expertise on virology and MM pathology (Dr. Carbone), AKT survival pathways (Dr. Testa) and use of normal human mesothelial and MM cells (Dr. Pass).

细胞外信号调节激酶 ERK1/2 和 ERK5 由间皮细胞中的石棉纤维激活 和气道上皮细胞，并在细胞存活中发挥关键作用。 ERK1/2 依赖性 Fra-1 表达也 与大鼠间皮细胞的形态转变和基因表达（c-met、cd44）有因果关系 刺激细胞增殖和迁移。我们假设 ERK1/2 和 ERK5 信号传导的激活 致癌纤维（石棉、毛沸石）在人类恶性肿瘤发病机制中发生的途径 间皮瘤 (MM) 会被 SV40 以共同致癌的方式增强。最近令人兴奋的数据也 表明这些生存途径在接触化疗药物后在 MM 中被激活，并且 可以通过操纵来实现增加细胞杀伤。因此，我们还假设 ERK1/2 和 ERK5 这些途径单独或协同地有助于化疗后 MM 细胞的存活。在目标 1 中，我们将测试 青石棉和温石棉、特征良好的土耳其毛沸石（参见项目 1）及其非纤维 类似物，单独或与 SV40 共同暴露，以确定 ERK1/2 和 ERK5 活性，fos/jun 家族 成员，AP-1 反式激活与转化模式和致癌性相关，如 由 Carbone 博士（核心 C）开发的体外/体内模型确定。在目标 2 中，我们将使用面板 来自 Core B 的 SV40+ 和 - MM 以确定显性失活结构和小发夹的影响 (sh) 针对体外细胞参数的 ERK1/2 和 ERK5 的 RNA 干扰 (RNAi) 载体 卡莫司汀 (BCNU) 治疗后的转化和生存。与 Testa 博士合作（项目 3) 我们还将确定这些研究中 AKT 生存途径是否被修改。在目标 3 中，一只老鼠 原位模型将用于确定 SV40+ 和 - MM 是否稳定转染 shMEKI、shERKS 或 在胸腔内注射和施用卡莫司汀之前，这两种结构都改变了生长和 转移。确定 MM 发病机制中细胞存活的途径对于 预防和治疗这些破坏性肿瘤。该计划项目允许我们以前的 啮齿动物间皮瘤模型的机制研究将在人类间皮瘤中得到验证，并提供 我们拥有病毒学和多发性骨髓瘤病理学（Carbone 博士）、AKT 生存途径（Testa 博士）和使用方面的专业知识 正常人间皮细胞和 MM 细胞 (Dr. Pass)。