Engineering Immuno-Glial-Neurovascular 3D-Brain-Chips with a Perfusable BBB for Accelerating Alzheimer’s Disease Drug Discovery and Translation

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PROJECT SUMMARY. While Alzheimer’s Disease (AD) is the sixth leading cause of death, there is still no pharmacologic treatment on the market that slows or stops neuronal damage in AD. Apolipoprotein-E4 (APOE4) is the strongest genetic risk factor for sporadic AD and one that roughly a quarter of the general population carries. Importantly, further precluding effective therapeutic development is the lack of human-based models that can recapitulate AD pathology with all the brain-resident cell types in the immune-glial-neurovascular unit, cap- turing critical cell non-autonomous effects, including in inflammatory and lipid dysregulation, and blood brain barrier (BBB)-mediated transport of nutrients and therapeutics. To address these limitations, in an ambitious moonshot project in the first phase of my postdoctoral work, I have engineered a novel brain-mimetic matrix (NeuroMatrix) that supports the co-culture of all 7 brain cell types from patient-specific induced pluripotent stem cells with mature phenotypes to form a multi-cellular integrated brain model (miBRAIN). This model recapitulates APOE4-associated dysregulation and AD pathological hallmarks of neuronal hyperexcitability, amyloid accumu- lation, phosphorylated tau burden, etc. Further, I have developed a novel constriction-minimizing microfluidic device that enables perfusable vasculature within neurovascular units self-assembled in NeuroMatrix. I propose here to: (Aim 1) leverage the miBRAIN platform to construct a diverse “in vitro patient cohort” across APOE genotype with isogenic lines to probe APOE4-specific effects and conduct important validation of the model as I aim to hone this platform technology for broad utility and (Aim 2) engineer 3D-miBRAIN- Chips by combining my novel microfluidic devices with the miBRAIN and successively integrating each cell type towards a fully perfusable BBB within miBRAIN culture and harness them to functionally assess BBB perme- ability and selectivity across patient cell lines and APOE status. After consultation with the Program Officer and given my long-term goals, I have decided to further focus on (Aim 3) investigating the best path forward for deploying this technology for maximal utility, including development of a business plan and identification of potential partners and focus applications. This work thus could result in important insights for AD mechanistic understanding, of immediate relevance to other neurodegenerative diseases associated with APOE4 risk, while developing a well-validated 3D-miBRAIN-Chip platform technology that is an integrated immune-glial-neurovas- cular unit with patient-specific genetics and biomimetic phenotypes and functions that could be of great utility for therapeutic development across CNS pathologies. Simultaneously, it provides vital training in research meth- ods (platform development, bioinformatics, lipidomic and inflammation analyses, mentoring from expert Co- Mentors and Scientific Advisory Committee) and career development (immersive entrepreneurial training, niche-specific coursework, mentoring from expert Co-Mentors and Entrepreneurial Advisory Committee, pursuit of ambitious independent vision) that will make a marked difference in launching my independent career.