MicroRNA control of the metabolic response to hypoxia in malignancy

该项目将进一步发展这些初步的方法，利用更先进的3D培养技术，包括球体和旋转细胞培养。这将使我们能够更深入地评估microRNA对包括缺氧在内的各种微环境条件的反应。激光显微切割将用于评估3D细胞培养和患者肿瘤标本中特定的肿瘤细胞定位和反应的异质性。将通知免疫组织化学在蛋白质水平上验证结果，使用已经构建的室内组织微阵列。作用也可以通过质谱学在代谢水平上进行分析。靶标的功能分析将通过microRNA或基因敲入/下调进行，增殖、迁移和侵袭分析已经在实验室使用。原位脑瘤模型也可用于在该项目的后期阶段对任何候选对象进行体内分析。我们将寻求改变所有已知的代谢途径抑制剂的用途，以测试是否可能作为GBM治疗的候选药物。

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.