A 3Rs approach to tumour metastasis: investigating the role of cancer stem cells in the metastasis of oral squamous cell carcinoma using an in vitro m

肿瘤转移的 3R 方法：使用体外模型研究癌症干细胞在口腔鳞状细胞癌转移中的作用

• 批准号: 2773020
• 金额: --
• 依托单位: Queen Mary University of London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2773020
• 关键词: 3Rs; approach; tumour; metastasis; investigating

A 3Rs approach to tumour metastasis: investigating the role of cancer stem cells in the metastasis of oral squamous cell carcinoma using an in vitro microfluidic model 300,000 cases of oral squamous cell carcinoma (OSCC) are diagnosed every year, and around 30% of patients exhibit metastatic spread. Metastasis is a complex process by which cancer cells travel to a secondary site to form a new tumour. Previous work has suggested that cancer stem cells (CSCs) are capable of driving metastasis by switching between epithelial and mesenchymal states. However, the role they play in the interactions with the vascular system, a key component of the tumour microenvironment (TME) recognised to greatly influence metastasis, remains unclear. Here, I used microfluidic devices, a promising new in vitro model, to study these interactions. I fabricated these devices using photolithography and soft lithography techniques and used them to co-culture endothelial cells with OSCC cells in a fibrin gel, to assess the interactions between these two cell types in 3d. After developing and optimising this OSCC metastasis-on-a-chip model, I uncovered a two-way communication between cancer cells and endothelial cells that greatly changes the behaviour of both cell types. Most interestingly, cancer cell invasion undergoes marked changes and is significantly reduced in the presence of endothelial cells. Imaging of the matrix shows that endothelial cells degrade and remodel the matrix when undergoing sprouting angiogenesis in the fibrin gel. This remodelled environment restricts OSCC cell migration in the matrix, and full epithelial-to-mesenchymal transition (EMT) is suppressed - instead invasion progresses through streaming of epithelial tumour cells through pre-existing tracks made by endothelial cells. Immunofluorescent staining of the cells in the invasive streams demonstrates features that have been associated with partial EMT in metastatic CSCs. These findings may represent the generation of a vascular niche that acts to modify the surrounding TME and, through this, influences the invasive CSC phenotype.

肿瘤转移的3Rs方法：利用体外微流控模型研究癌症干细胞在口腔鳞状细胞癌转移中的作用，每年有30万例口腔鳞状细胞癌（OSCC）被诊断出来，约30%的患者表现出转移扩散。癌细胞转移是一个复杂的过程，癌细胞转移到继发部位形成新的肿瘤。先前的研究表明，癌症干细胞（CSCs）能够通过在上皮和间充质状态之间切换来驱动转移。然而，它们在与血管系统的相互作用中所起的作用仍不清楚，血管系统是肿瘤微环境（TME）的一个关键组成部分，被认为对转移有很大影响。在这里，我使用微流体装置，一个有前途的新的体外模型，来研究这些相互作用。我使用光刻和软光刻技术制造了这些设备，并使用它们在纤维蛋白凝胶中共同培养内皮细胞和OSCC细胞，以3d方式评估这两种细胞类型之间的相互作用。在开发和优化这个OSCC芯片转移模型后，我发现癌细胞和内皮细胞之间的双向交流极大地改变了两种细胞类型的行为。最有趣的是，癌细胞的侵袭发生了显著的变化，并且在内皮细胞的存在下显著减少。基质成像显示内皮细胞在纤维蛋白凝胶中进行新生血管生成时降解和重塑基质。这种重塑的环境限制了OSCC细胞在基质中的迁移，并且完全的上皮-间质转化（EMT）被抑制-取而代之的是，侵袭过程通过上皮肿瘤细胞通过内皮细胞预先存在的轨迹流动进行。侵袭性流式细胞的免疫荧光染色显示转移性CSCs中与部分EMT相关的特征。这些发现可能代表了血管生态位的产生，其作用是改变周围的TME，并通过这一点影响侵袭性CSC表型。