近年发现的肿瘤非内皮依赖性"血管"(血管拟态，VM)形成机制不清，缺乏阻抑措施，导致抗肿瘤血管生成疗效欠佳。我们前期研究发现，肿瘤干细胞(CSCs)不仅诱导血管生成、转分化为内皮，还是VM形成的关键细胞，其中VEGFR-2和Twist1可能起重要作用。本项目围绕"CSCs在VM形成中的作用及机制"这一关键科学问题，以人脑胶质瘤CSCs及其移植瘤为主要材料，创建一系列体外和在体VM研究模型，采用活细胞示踪、双光子与共聚焦显微术、显微切割与组学检测等技术，进一步研究VM特征、形成规律和细胞来源，明确CSCs表型转化(标志物)和归宿，认识VM形成中CSCs"干性"维持、基质分泌相关的主要信号通路，并针对VM的分子靶点，筛选拮抗剂、抑制剂或制备单抗，观测干预效应。研究结果对阐明肿瘤VM形成的细胞与分子机制及其诊断价值具有重要意义，并将为基于CSCs的抗VM个体化肿瘤治疗提供新策略、新方法。

Not only the lack of comprehensive understanding of the underlying mechanism for the formation of newly discovered vasculogenic mimicry (VM), a special type of microvessels lined by cancer cells other than vascular endothelia, but also none of effective inhibitory regimens to specifically target VM available for clinicians are major reasons that current conventional anti-angiogenesis therapies are less effective and even more often failed clinically. Our previous studies demonstrate that cancer stem cells (CSCs) not only contribute to angiogenesis but also give rise to endothelial cells and participate in the VM formation of which VEGFR-2 and Twist1 may play an important role. This study will focus on investigating the roles and mechanisms of CSCs in the formation of VM. Using CSCs derived from glioma to establish several in vivo and in vitro new models for VM study, we will employ several technologies available in our group including live cell tracking, two-photon and laser confocal microscopy, laser capture microdissection, and all "-omic" analyses, etc., not only to investigate VM characteristics, development and cellular origin, but also to reveal its phenotype transition (surface markers) and destination, and identify the signaling pathways that control the stemness maintenance and extracellular matrix production in CSCs during the process of VM formation. Finally, we will try to find or screen for inhibitors, agonists and/or antibodies specific for VM-related molecules and/or signaling pathways and examine their efficacies for intervention. The results of this proposed study is expected not only to provide a better understanding for cellular and molecular mechanisms of VM formation and demonstrate its important clinical significance for prognostic evaluation and more accurate pathological diagnosis of cancer, but also to yield novel strategies and regimens for CSCs-based personalized VM-targeted anti-tumor therapy to treat cancer patients.