基于微流控芯片的肿瘤微环境中细胞通讯机制的研究

Tumor cells release excessive amount of exosomes and micro- vesicles, which may influence tumor growth and progression. However, a further verification is still required. Previously, we found that the fate of tumor cell was depended on the location of tumor suppressor gene (ONCOL REP). Then, we designed and fabricated a multilayer (3D), multi-cell (multicomponent) and multi-functional microfluidic organ-like chip for drug screening, providing a controllable culture microenvironment for living cells in micrometer-sized chambers to simulate physiological functions of tissues and organs(SCI RE). In this project, we try to develop a microfluidic system that mimics the tumor microenvironment containing various cells. This long-term maintainable system well facilitate observation on the effect of manual interfered extracellular vesicles in tumor progression in the condition of RAB27A (required for vesicles secretion) and Lnc H19 (metastasis)knock-in or -out . The changes of components (protein and RNA) in the vesicles along with tumor progress and the key factors should be revealed by MS-based proteomics and nucleic acid sequencing analysis. Based on Brca1F22-24/F22-24；p53+/- spontaneous breast cancer mices and existing compound libraries, we plan to verify the relationship between vesicles and tumor organ-specific metastasis and select a vesicle secretion inhibitor, which keeps potential to be an anti-tumor drug.

肿瘤细胞能分泌大量外泌体与微囊泡影响肿瘤的增殖与转移，但尚需验证。申请人此前发现抑癌基因不同的定位直接影响肿瘤细胞的状态(ONCOL REP)，并构建了集成多种器官和组织的类器官芯片，在体外微米尺度的腔室中，创造可控的细胞培养环境，模拟组织或器官的生理状态及功能(SCI RE)。因此，本项目设计利用微流控技术构建具有多种细胞的肿瘤微环境模型，通过长期培养观察敲除或高表达RAB27A(囊泡分泌所需基因)、lncRNA H19(转移相关)对肿瘤发展的影响，利用质谱-蛋白质组学与RNA测序的方式研究在肿瘤微环境模型在发展过程中分泌的囊泡内蛋白与RNA的变化规律，以及作为临床检测标记物的潜力；利用已有的小分子文库筛选能抑制囊泡分泌的化合物，并通过BLG-Cre；Brca1F22-24/F22-24；p53+/-转基因自发乳腺癌小鼠模型验证囊泡与肿瘤定向转移的关系及作为治疗靶点的潜力。