ERK5促进黑色素瘤对威罗菲尼耐药的作用和机制

项目名称： ERK5促进黑色素瘤对威罗菲尼耐药的作用和机制

项目编号： No.81502622

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 王丽娜

作者单位： 大连医科大学

项目金额： 18万元

中文摘要： 黑色素瘤恶性程度高，BRAF抑制剂威罗菲尼已广泛用于其治疗，但耐药性仍是治疗瓶颈。申请人前期利用SCAM等技术，阐述了RAS上游组氨酸蛋白激酶（HPK）经拓扑结构改变产生耐药的分子机制(Biochim Biophys Acta, 2014)。预实验发现对威罗菲尼耐药的黑色素瘤细胞中ERK5通路持续激活，且ERK5抑制剂XMD8-92与威罗菲尼联合使用显著抑制肿瘤。据此提出假说，耐药黑色素瘤细胞中ERK1/2抑制，ERK5表达上调且持续激活，进而导致其对威罗菲尼耐药。本课题拟利用RNA-seq、过表达和Cas9介导基因敲除等手段，通过检测XMD8-92和威罗菲尼等处理下，非耐药和耐药细胞增殖等变化，结合小鼠移植瘤模型，研究ERK5在黑色素瘤对威罗菲尼耐药中的作用；同时构建受ERK5通路调控的基因表达图谱，进行表达检测，并结合功能分析，探讨其耐药机制和作用靶点，为改善临床化疗效果提供新思路。

中文关键词： 黑色素瘤；XMD8-92；细胞外信号调节激酶5；耐药机制；；威罗菲尼

英文摘要： Melanoma is characterized by high malignancy. Vemurafenib that targets the BRAF kinase has been widely used in treating melanoma, however, drug resistance is still the bottleneck of treatment. Using substituted cysteine accessibility method (SCAM), we described that the resistance mechanism of the RAS upstream, histidine protein kinase (HPK) by alteration of the topology structure (Biochim Biophys Acta, 2014). The pilot experiment indicated that extracellular signal regulated kinase (ERK5) pathway is universally activated in melanoma after the development of resistance to BRAF inhibitors, vemurafenib. And ERK5 inhibitor, XMD8-92, in combination with vemurafenib significantly inhibited the tumor growth. Based on these, it is hypothesized that ERK1/2 is suppressed and ERK5 is activated in vemurafenib-resistant melanoma . In this study, RNA-seq, overexpression and Cas9-mediated knock out will be applied. After the specific treatment by small molecular dugs that target the ERK1/2 and ERK5 pathways, proliferation of the non-resistant and resistant cells will be investigated. Mouse model will also be applied to study the role of ERK5 in vemurafenib-resistant melanoma. Furthermore, the gene expression map regulated by the ERK5 pathway will be built to unveil the chemo-resistant mechanism of melanoma. We hope that this study could provide a new insight into improving clinical efficacy.

英文关键词： Melanoma;XMD8-92; Extracellular signal regulated kinase (ERK5) ; Resistance mechanisms;Vemurafenib