MicroRNA control of the metabolic response to hypoxia in malignancy

该项目将进一步开发这些初始方法，利用更先进的3D培养技术，包括球体和旋转细胞培养。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。激光显微切割将用于评价特定的肿瘤细胞小生境，并评估3D细胞培养和患者肿瘤标本中反应的异质性。将通知免疫组织化学，以使用已构建的内部组织微阵列在蛋白质水平上验证结果。还可以通过质谱法在代谢水平上分析影响，通过microRNA或基因敲入/敲低进行靶点的功能分析，实验室中已经使用增殖、迁移和侵袭测定。原位脑肿瘤模型也可用于在项目后期对任何候选人进行体内分析。我们将寻求重新利用任何已知的代谢途径抑制剂，以测试作为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.