调控网络研究T细胞在微泡促进癌变中的功能转变机制

T cells are very important in tumor immunology and they can suppress or promote tumorigenesis. The functional transformation of T cells is also a key issue in immunotherapy. We have developed a leukemogenesis cell model with microvesicles derived from K562 cells and studied its regulatory network and mechanism. Our preliminary experiments found that T cells have undergone a functional transformation process from activation to repression in the microvesicle induced leukemogenesis model under unknown regulatory mechanism. Based on this leukemogenesis model, preliminary experiments and our regulatory network analysis, in this project, we will study the regulatory mechanism of T cell functional transformation utilize the transcription factor (TF) and miRNA regulatory network analysis. We will analyze the expression changes of gene and miRNA in T cells, design algorithm to infer the variation of T cell receptors, construct TF and miRNA co-regulatory network for genes in T cells. Then, we will develop methods to integrate multiple data to screen key regulatory modules and regulators for T cell functional transformation in the network and verified them by experiments. Further to discuss their potential application possibility as regulators for T cell functions. In this project, we used unique models and bioinformatics methods to explore the T cell functional transition in tumorigenesis. It will provide basic knowledge for improving the efficacy of immunotherapy, which will have important theoretical significance and potential application values.

T细胞在肿瘤免疫中非常重要，可抑制或促进肿瘤发生，其功能转变是免疫治疗中的关键问题。我们建立了K562微泡诱导白血病发生模型，研究了其调控网络和机制。预实验发现T细胞在该癌变过程中经历了由激活到抑制的功能转变，具体调控机制不清楚。基于该癌变模型、预实验和调控网络等研究基础，我们将从转录因子与miRNA调控网络研究T细胞在癌变中功能转变的调控机理。分析癌变中T细胞基因和miRNA表达的差异，设计算法推断T细胞受体的变化，构建转录因子和miRNA共调控T细胞基因表达的调控网络，开发方法整合多种信息筛选网络中的关键调控模块和分子，发现对T细胞功能转变有重要影响的调控因子和模块并实验验证，讨论其作为T细胞功能调节分子的潜在应用可能性。本项目利用独有模型和生物信息方法，探讨T细胞在癌症发生中的变化和调控机制，为解析T细胞在癌变中的作用和提高免疫治疗疗效提供研究基础，有重要的理论意义和应用价值。

细胞外囊泡（EVs）是细胞通讯的重要组成部分。本课题主要围绕细胞外囊泡和miRNA调控开展。针对获取的T细胞经细胞外囊泡（EVs）诱导后状态改变过程的样本进行RNA-seq测序。分析T细胞状态转变过程的基因表达和miRNA表达差异，探究了EV和T细胞之间的交流，并构建了转录因子调控网络，筛选了核心调控分子和通路，首次发现EV诱导T细胞耗竭的现象并进行了初步机制探索与验证。同时，围绕EV，我们研究了骨髓瘤细胞外囊泡（MM-EV）的生物活性成分，经分析检测，确认了MM-EV抑制骨髓间充质干细胞（BM-MSC）的成骨功能以及促进骨髓瘤骨病的影响。通过miRNA测序分析发现，MM-EV中的miRNA包括miR-103a-3p可能与成骨抑制作用有关。发现骨髓瘤骨病人血液中CD138+ 循环EV计数与骨病变的数量有关，表明其可能是骨髓瘤骨病的潜在诊断标志物。.我们还构建了miRNA相关SNP数据库miRNASNP3.0，长非编码RNA相关SNP数据库lncRNASNP2.0，并建立了基因在不同组织、疾病和细胞系中的表达查询和现实平台GEDS。分析了CAR-T治疗过程中基因表达变化及其调控网络，发现一些与治疗疗效相关的关键分子。