Understanding Neuroinflammation through advanced in vitro 3D modelling using human Pluripotent Stem Cells

Microglia - brain-resident macrophages - carry out homeostatic surveillance functions in the brain, clearing dying cells, extracellular debris, and pruning synapses during development. However, build-up of debris, especially aggregated proteins, causes chronic microglial activation, contributing to neurodegeneration and psychiatric disorders. Microglial inflammatory damage may be direct, or mediated by astrocytes, which normally support neuronal metabolism but become damaging under instruction from microglia. Clearly, understanding the normal function of microglia, their interactions with neurons and astrocytes, and the mechanisms by which this triumvirate becomes perturbed, is crucial to our understanding of brain health, mental health and diseases of ageing - and authentic human cellular models are necessary to achieve this.
Our team have pioneered the generation of macrophages and microglia from human induced Pluripotent Stem Cells, and worked extensively with iPS-neuronal models. However, the 2D models we work with currently do not form mature, extensive synaptic networks or develop the pathological features of protein aggregation. In 3D culture, neurons mature further and aggregated protein pathology can develop. This D.Phil project will develop and exploit a relatively simple 3D triculture system, containing iPS-microglia, astrocytes and cortical neurons. The research project will expose the student to cutting-edge cell and molecular biology, imaging, biochemistry and sequencing technologies. A suite of neuronal, astrocytic and microglial assays will be deployed to assess cellular function and maturity, and for full exploitation of the 3D system, several advanced imaging methodologies can be employed, including confocal, multiphoton and Lightsheet microscopy, InCell and Opera Phoenix high-content confocal imaging systems.
The student will be primarily based at the academic partner institution, the Sir William Dunn School of Pathology, University of Oxford, with secondments to the industrial partner, Eli Lilly, as necessary. Eli Lilly is geographically very close (1 hour travel from Oxford), so the student can be seconded to the industrial partner very flexibly, according to the research progress, for periods of 1-3 weeks at a time, and for a minimum of 3 months in total across the 4 year D.Phil project. The Lilly Research Centre in Windlesham has modern biochemistry (automated liquid handling systems, AlphaLISA, MSD, SiMoA, automated Western blot systems, DLS, Flow cytometry), cell biology (Opera and Operetta Phoenix high-content imaging systems) and molecular biology (Nanostring, RT qPCR) labs, in which the student will receive appropriate professional training in support of the project work.