基于单细胞质谱流式技术研究肌层浸润性膀胱癌分子亚型异质性以及KLF14介导的Ba/Sq亚型免疫抑制微环境形成机制

肌层浸润性膀胱癌（MIBC）是一组具有不同临床病程、不同治疗反应且机制尚未明确的分子异质性肿瘤。基于不同基因表达模式的分子分型策略对于阐明MIBC的机制提供了新的框架。本项目前期采用最新的共识性分子分型法分型鉴定MIBC，利用质谱流式技术从单细胞层面初步阐明六种亚型肿瘤干细胞类亚群和免疫细胞亚群异质性特征，其特征细胞亚群可能与患者的预后相关。进一步研究发现Ba/Sq亚型是免疫抑制程度最高的亚型，其机制可能与转录因子KLF14对调节性T细胞的功能调控有关。本项目拟采用质谱流式、镜像质谱流式和全转录组测序技术，多维度精准分析MIBC各亚型肿瘤干细胞类亚群和免疫细胞亚群，找出与临床预后相关的特征细胞亚群，阐明其基因表达特征，并结合CRISPR/Cas9基因编辑技术研究KLF14对Ba/Sq亚型免疫抑制微环境形成的作用和机制，本研究有助于开发新的个性化治疗方案以改善MIBC患者的生存质量。

Muscle invasive bladder cancer (MIBC) is a group of molecular heterogeneous tumors with different clinical course, different treatment responses, and the mechanism is not yet clear. Molecular classification strategies based on different gene expression patterns provide a new framework for elucidating the mechanism of MIBC. In the previous stage of this project, MIBC were identified by the latest consensus molecular classification strategy. The heterogeneity characteristics of tumor stem-like cell subsets and immune cell subsets of six subtypes were preliminarily elucidated from the single cell level by mass cytometry, and their characteristic cell subsets may be related to the prognosis of patients. Further study found that Ba/Sq subtype is the most immunosuppressive subtype, and its mechanism may be related to the regulation of regulatory T cell function by transcription factor KLF14. The purpose of this project is to use mass cytometry, imaging mass cytometry and RNA-seq techniques to accurately analyze tumor stem-like cell subsets and immune cell subsets of MIBC subtypes from multi-dimensional, to identify characteristic cell subsets related to clinical prognosis and clarify their gene expression characteristics, then to combine CRISPR/Cas9 gene editing technology to study the effect and mechanism of KLF14 on the formation of immunosuppressive microenvironment of Ba/Sq subtypes. This study is helpful to develop personalized treatment schemes to improve the quality of life of patients with MIBC.