Project - Alzheimer’s Disease Insights and Drug Development using a 3D iPSC-Derived Triculture System (2019-2022)
Research Project Description
The lack of effective therapeutics for AD is partly due to the lack of understanding of AD pathology and the lack of translatable screening/validation platforms for potential therapeutics. One factor that will facilitate a more in-depth understanding of AD pathology is the ability to create in vitro models that effectively recapitulate all aspects of the in vivo disease state. A multitude of techniques have been previously used to model the AD environment. Both in vitro and in silico techniques have significantly contributed to our current understanding. Recently, 3D culture techniques and efficient protocols for iPSC differentiation have provided significant breakthroughs in our understanding of AD pathology. Specifically, co-culture of neurons, astrocytes and microglia have the potential to provide new insights into how cell-to-cell communication impacts specific pathologies of AD and neurodegeneration. iPSC cultures both in isolation and co-culture have shown to provide multiple benefits in modelling AD.
Utilising both 3D culture and iPSC derived astrocytes and microglia from both control and familial AD patients, the project will provide insights into glial cell dysfunction and potential novel therapeutic pathways. Additionally, there is no 3D tri-culture system for AD using iPSC-derived neurons, astrocytes and microglia. The technology being used to create the 3D cell culture environment provides the current project with the ability to uniquely create a 3D environment autonomously and potentially screen therapeutics in a 96-well medium-to-high throughput manner.
The relevance of this project lies in the demonstration of physiologically relevant glial pathophysiology and neuron-glial interactions: glial morphogenesis, recruitment, phagocytosis, inflammatory cytokine/chemokine secretion and global neuronal death. This project will allow a deeper understanding of AD pathology, allowing for novel target identification and a platform for pre-clinical screening/evaluation of AD therapeutics.
A variety of methods will be used within this project, including 3D cell bio-printing, the development of a novel hydrogel to sustain CNS cell cultures, ELISA’s, immunofluorescence imaging, iPSC differentiation and CRISPR gene editing.