(W1340) In Vitro Human Neurovascular Unit Model Using Cells Differentiated from a Single iPSC Lineage for Metabolic and Neurodegenerative Diseases Research
Full Professor UNIFESP São Paulo, Sao Paulo, Brazil
Abstract: The neurovascular unit (NVU) is the functional interface between the vascular system and the brain, coordinating communication among neurons, glial cells, and blood vessels. It is vital for brain homeostasis, and its dysfunction is linked to metabolic and neurodegenerative diseases such as diabetes, obesity, and Alzheimer’s disease. Most NVU studies rely on animal models or mixed human-animal systems, which can limit its application in human diseases. To address this, we developed a human NVU model comprising endothelial cells (iEC), astrocytes (iAST), and neurons (iNeu), all differentiated from the same iPSC line. Human iPSCs were differentiated into iEC, iAST, and iNeu, characterized for functionality, and optimized for culture conditions using tailored media and cellular densities. The three cell types were co-cultured in a transwell system to mimic the human NVU (hNVU). The model was validated via permeability assays, TEER measurements, and fatty acid transport tests. The hNVU exhibited high TEER values (>2000 Ω·cm²) and low permeability ( < 4% FITC-Dextran paracellular transport). Cells expressed key proteins and transporters, such as MFSD2A, P-gp, FABP5, FABP7, GLUT1, and CD36. Additionally, iECs internalized and transported fluorescent fatty acid analogs (BODIPY FL C16 and LysoPC-TopFluor). The model's applicability for metabolic and neurodegenerative research was evaluated by exposing iECs to hyperglycemia (15 mM glucose + 100 µg/mL AGE-BSA) or 200 nM oligomeric human β-amyloid (1-40) for 48 hours. Both treatments disrupted tight junction proteins (ZO-1 and occludin) and altered transporter expression (GLUT1 and MFSD2A) with β-amyloid clustering and aggregation. In conclusion, the hiPSC-derived NVU model closely mimics in vivo conditions, offering a human-specific platform for studying metabolic and neurodegenerative diseases. Using a single iPSC line for all cell types enhances physiological relevance and enables a personalized approach.
