MicroRNA control of the metabolic response to hypoxia in malignancy

该项目将进一步发展这些最初的方法，利用更先进的3D培养技术，包括球形和旋转细胞培养。这将允许更深入地评估microRNA对不同微环境条件（包括缺氧）的反应。激光显微解剖将用于评估特定的肿瘤细胞龛和评估反应的异质性，无论是在3D细胞培养和患者肿瘤标本中。免疫组织化学将被告知在蛋白质水平上验证结果，使用已经构建的内部组织微阵列。效果也可以在代谢水平上通过质谱分析。靶标的功能分析将通过microRNA或基因敲入/敲低进行，增殖、迁移和侵袭试验已经在实验室中使用。原位脑肿瘤模型也可用于在项目后期对任何候选肿瘤进行体内分析。我们将寻求重新利用任何已知的代谢途径抑制剂，作为GBM治疗的可能候选药物。

The project will develop these initial approaches further, utilising more advanced 3D culture techniques, including spheroid and rotary cell culture. This will allow more in depth assessment of the microRNA response to varying micro-environmental conditions, including hypoxia. Laser microdissection will be used to evaluate specific tumour cell niches and assess heterogeneity of the response, both in 3D cell culture and in patient tumour specimens. Imunohistochemistry will be informed to validate results at a protein level, using already constructed in house tissue microarrays. Effects can also be analysed at a metabolic level by mass spectrometry.Functional analysis of targets will be undertaken by microRNA or gene knock in/down, with proliferation, migration and invasion assays already in use in the laboratory. Orthotopic brain tumour models are also available to allow in vivo analysis of any candidates at the late stages of the project. We will seek to repurpose any known inhibitors of the metabolic pathways interrogated to test as possible candidate agents for GBM therapy.