HPV16整合源性外泌体c-Myc激活TREM-2+巨噬细胞免疫抑制功能促进宫颈上皮内瘤变进展的机制研究

高危HPV整合诱导的免疫逃逸在CIN进展中发挥重要作用，其调控机制尚不明确。在前期证实HPV16整合与巨噬细胞表型及功能改变密切关联的基础上，本项目预实验显示HPV16高频整合位点基因产物c-Myc以外泌体转载的形式促进TREM-2+巨噬细胞极化并激活其对效应T细胞的免疫抑制功能；生物信息学预测TREM-2结合DAP12参与T细胞功能调控的下游通路是RAS-MAPK，正性调控Arg1表达。由此我们提出科学假说：HPV16整合源性外泌体c-Myc作用于巨噬细胞，通过TREM-2/DAP12/RAS-MAPK/Arg1信号轴重塑宫颈局部免疫抑制微环境，促进CIN进展。本项目拟利用高通量病毒整合检测、外泌体示踪、CRISPR/Cas9-sgRNA全基因组文库筛选、HPV16转基因CIN小鼠模型等关键技术，探讨高危HPV整合相关CIN进展的免疫逃逸机制，为临床早期预测和阻断CIN进展提供新靶点。

High-risk HPV integration-induced immune escape plays an important role in the progression of cervical intraepithelial neoplasia (CIN). However, the underlying mechanism is still elusive. On the basis of previous studies confirming that HPV16 integration was closely related to phenotypic and functional changes of macrophages, our preliminary experiment further demonstrated that HPV16 high- frequency integration site gene product c-Myc was transferred into macrophages in the form of exosome, promoting the polarization of TREM-2+ macrophages and activating its immunosuppressive function against effector T cells. Bioinformatics predicts that the downstream pathway of TREM-2 combined with DAP12 involved in the regulation of T cell function is RAS-MAPK, which positively regulates the expression of Arg1. Therefore, we propose that HPV16 integration-derived exosomal c-Myc acts on macrophages to reshape the local cervical immunosuppressive microenvironment through TREM-2/DAP12/RAS-MAPK/Arg1 signal axis and promotes the progression of CIN. This study intends to use several key technologies, such as high-throughput viral integration detection, exosome tracking, genome-wide library screening of CRISPR/Cas9-sgRNA, HPV16 transgenic CIN mouse model, etc., to explore the immune escape mechanism of high-risk HPV integration-related CIN progression, which will provide scientific evidence for exploring early prediction and effective therapeutic targets of inhibiting the progression in CIN.