基于MIF外泌体-Jab1/JNK/AP1信号通路探讨双参颗粒重塑TAMs介导的Kras突变肺癌肿瘤免疫微环境的机制研究

The prevention and treatment of Kras gene mutation in lung cancer is to be solved urgently. One of the important features of lung cancer patients with Kras gene mutation is the recruitment of immune cells to form a tumor immunosuppressive microenvironment. Tumor-associated macrophages (TAMs) are important components of the tumor immunosuppressive microenvironment which can promote tumor immune escape and progression. The MIF exosome-Jab1/JNK/AP1 pathway is an important pathway for promoting TAMs to mediate tumor immunosuppression microenvironment. Shuangshen Granule is a prescription developed by Oncology Department of Guang'anmen Hospital which has been confirmed effective in the prevention and treatment of lung cancer. This study will construct a mouse model of spontaneous lung cancer which can be induced by KrasG12D mutation and a tumor-immune cell co-culture model of KrasG12D mutation. And through different techniques such as tissue multi-spectral imaging, high content analysis, RT-qPCR, Western-blot, ELISA, flow cytometry and other techniques, we would like to explore the molecular mechanisms of how Shuangshen granules remodel the immunophenotype of TAMs and reverse the tumor immunosuppression microenvironment, and reveal the new connotation of Yiqi Yangyin Huoxue method in preventing and treating lung cancer. We hope that this study could provide new scientific basis in further new drug development in preventing lung cancer.

Kras基因突变肺癌的防治是目前亟待解决的临床难题，招募免疫细胞形成肿瘤免疫抑制微环境是影响kras基因突变肺癌临床疗效的的重要因素之一。肿瘤相关巨噬细胞（TAMs）是肿瘤免疫抑制微环境的重要组成部分，能够促进肿瘤的免疫逃逸和进展转移。MIF外泌体-Jab1/JNK/AP1通路是促进TAMs介导肿瘤免疫抑制微环境的重要通路。双参颗粒是广安门医院肿瘤科防治肺癌的经验集萃，前期研究证实其可以防治肺癌的复发转移。本课题拟通过构建KrasG12D突变自发肺癌小鼠模型和肿瘤-免疫细胞共培养模型，以MIF外泌体-Jab1/JNK/AP-1通路为切入点，运用组织多光谱成像、高内涵分析、RT-qPCR、Western-blot、ELISA、流式细胞等技术，探讨双参颗粒重塑TAMs免疫表型和逆转肿瘤免疫抑制微环境的分子机制，揭示益气养阴活血法防治肺癌的新内涵，为新药创制和肺癌防治提供新的科学依据。

Kras基因突变肺癌的防治是目前亟待解决的临床难题，招募免疫细胞形成肿瘤免疫抑制微环境是影响kras基因突变肺癌临床疗效的的重要因素之一。肿瘤相关巨噬细胞（TAMs）是肿瘤免疫抑制微环境的重要组成部分，能够促进肿瘤的免疫逃逸和进展转移。MIF外泌体-Jab1/JNK/AP1通路是促进TAMs介导肿瘤免疫抑制微环境的重要通路。双参颗粒是广安门医院肿瘤科防治肺癌的经验集萃，前期研究证实其可以防治肺癌的复发转移。本研究发现，1、双参颗粒抑制KrasG12D突变自发肺癌小鼠模型和肺癌荷瘤小鼠模型肿瘤生长；2、双参颗粒抑制M2型肿瘤相关巨噬细胞（TAMs）表达，促进M1型TAMs表达；3、双参颗粒调控Treg细胞和肿瘤记忆T细胞表达；4、双参颗粒调控和抑制肺癌肿瘤TAMs相关因子iNOS、ARG1、TGF-β、VEGF、MMP-9的蛋白表达；5、双参颗粒抑制肺癌肿瘤细胞MIF蛋白表达；6、双参颗粒促进肺癌外泌体MiR-34a表达而调控TAMs表型；7、双参颗粒调控外泌体分泌相关因子PKM2、SNAP23、miR-146a及TAMs 细胞相关通路因子KLF4、Jab1、JNK、AP1的表达水平。本项目揭示了益气养阴活血法抗肺癌的新的科学内涵，为进一步的新药创制奠定了基础。发表核心期刊论文2篇，SCI论文2篇，申请国家发明专利1项，培养硕博研究生各1人。