This project will build important new tools for understanding the role of the immune system in amyotrophic lateral sclerosis (ALS), a major form of motor neuron disease. Inflammatory responses by resident brain and spinal cord immune cells (microglia) have an important role in ALS and are key targets for therapy. Until now, research on microglia has been largely restricted to cells from animal origin, and grown in 2 dimensional (2D) cultures. We now have new techniques to generate and grow microglia directly from ALS patients in 3D to help understand the disease and test patient-specific drugs to modulate the immune response in the brain and spinal cord. This project will provide a new approach to investigating and treating inflammation in ALS.
In this project we will simultaneously investigate two new complementary approaches to generate ALS patient microglia. Both approaches involve recently developed techniques that have yet to be applied to ALS research. Monocyte-derived microglia provide a relatively easy and rapid approach to examine microglia-like cells from larger numbers of ALS patients in ‘real-time’. Microglia derived from ALS patient induced pluripotent stem cells (iPSCs) is a relatively more time consuming approach but has the advantage of generating microglia through the correct ontogeny (origin and developmental pathway) producing a highly accurate representation of adult human brain-derived microglia. These approaches are highly complementary. The monocyte-derived microglia approach is ideal for rapidly investigating patient-specific therapeutics, and the iPSC-derived microglia are ideal for in-depth studies on the role of microglia in ALS onset and progression.
We will generate monocyte and iPSC-derived microglia from ALS patients and examine the inflammatory response compared to matched controls. The cells will be characterized for response to inflammatory stimuli including cytokine response, phagocytic activity, microglial ramification and surveillance, and response to inflammatory modulating compounds. Gene expression analysis will also provide valuable predictive information that may allow rapid assessment of potential therapeutic neuroinflammatory drug responses from each patient gene expression profile (personalized medicine). Microglia will also be grown in 3D OrganoPlateTM microfluidic platforms together with iPSC-derived spinal cord neurons and astrocytes to provide a unique insight into neuroinflammatory interaction between microglia and other key brain cell-types. The unique 3D model system will provide an ideal localized environment that more closely represents the complex 3D milieu of the brain and spinal cord, which is critical to understand microglia action, a cell-type highly sensitive to the localized environment.
This project will provide a major advance in the development of new approaches to understand microglia involvement in ALS and ‘proof of concept’ that such a system can be used to identify potential therapeutic agents in real-time. Techniques will include microglia and neural stem cell culture, molecular studies (i.e. RT-PCR), microscopy (confocal imaging), various biochemical assays (i.e. cytokine bead arrays) and protein analysis (western blot).
- PhD project but may also be considered for an Honours project.