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TRACER: a 3D engineered tumour for mapping cell metabolism and phenotype in heterogeneous microenvironments

机译:TRACER:3D工程化肿瘤,可绘制异质微环境中的细胞代谢和表型

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In recent years, emphasis has been placed on developing in vitro platforms that recapitulate the native tissue microenvironment to enable more effective compound screening compared with 2D culture. Though many systems - especially cancer models - have been engineered with physiological complexity, rarely has emphasis been placed on modes of data collection - limiting their applications. Here we describe an engineered tumour developed with data retrieval in mind. The model is assembled through single step rolling of a scaffold-tumour cell composite strip into a layered 3D coil. The model can be rapidly disassembled by unrolling for snapshot data analysis of tumour microenvironment in sync with cell phenotype in each of the layers. The model mimics the region of tumour tissue adjacent to a blood vessel, and recapitulates growth variations, gene expression profiles, and therapy responses consistent with tumours in vivo. Using our model, we show that the establishment of oxygen gradients are shaped by oxygen dependant signalling pathways, and use liquid chromatography tandem mass spectrometry to map cellular metabolism in 3D, concomitant to oxygen gradient. Our analysis identifies spatially defined metabolic signatures of cancer, revealing both known and novel metabolic responses to hypoxia.
机译:近年来,重点已放在开发体外平台上,该平台概述了天然组织的微环境,与2D培养相比,能够进行更有效的化合物筛选。尽管许多系统(尤其是癌症模型)的设计具有生理复杂性,但很少强调数据收集的方式-限制了它们的应用。在这里,我们描述了一种考虑到数据检索的工程肿瘤。通过将脚手架-肿瘤细胞复合材料条单步轧制成分层的3D线圈来组装模型。通过展开与模型的每一层中的细胞表型同步的肿瘤微环境快照数据分析,可以快速拆卸模型。该模型模拟与血管相邻的肿瘤组织区域,并概括与体内肿瘤一致的生长变异,基因表达谱和治疗反应。使用我们的模型,我们表明氧梯度的建立受氧依赖性信号传导途径的影响,并使用液相色谱串联质谱法在3D中伴随氧梯度映射细胞代谢。我们的分析确定了癌症在空间上定义的代谢特征,揭示了对缺氧的已知和新型代谢反应。

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