Fully Human Stem Cell-Derived BBB Model for Clinical Disease Modeling and Neurotropic AAV selection

Iannielli, A.; Giannelli, S. G.; Behrens, T.; Luoni, M.; Muggeo, S.; Moscato, G.; Greco, M.; Lenzi, E.; El Khoury, R.; Criscuolo, E.; Casucci, M.; Clementi, N.; Canals, J. M.; Iorio, R.; Broccoli, V.

doi:10.64898/2025.12.21.695793

bioRxiv · 10.64898/2025.12.21.695793

Fully Human Stem Cell-Derived BBB Model for Clinical Disease Modeling and Neurotropic AAV selection

Iannielli, A.·Giannelli, S. G.·Behrens, T.·Luoni, M.·Muggeo, S.·Moscato, G.·Greco, M.·Lenzi, E.·El Khoury, R.·Criscuolo, E.·Casucci, M.·Clementi, N.·Canals, J. M.·Iorio, R.·Broccoli, V.

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Abstract

The blood-brain barrier (BBB) is a highly functionalized vascular interface which regulates the exchange of substances between the neural parenchyma and its periphery. BBB leakage, leading to its uncontrolled permeability, is increasingly recognized to facilitate the onset of neuropathologies and aggravate their clinical progression. In vitro models of the BBB have rapidly evolved into elaborated structures that mimic its spatial architecture and multicellular nature. However, their cellular components are currently highly heterogeneous in origin and maturation state. Here, we have developed novel procedures to establish reproducible and scalable sources of endothelial, mural and astroglial cells generating a fully human pluripotent stem cell (hPSC)-derived BBB model, termed thBBBA. hPSC-derived BBB cell types are readily assembled into thBBBAs that develop mature functional properties with high barrier impermeability. Mature thBBBAs can also be generated by frozen hPSC-derived cell samples, providing a simple and scalable off-the-shelf system for general use. thBBBAs were instrumental in identifying the critical pathological role of an IL-6 autocrine source in disrupting thBBBA integrity, increasing its permeability to NMDAR antibodies from autoimmune encephalitis patients, and revealing the therapeutic effects of tocilizumab in this setting. Additionally, we have shown that thBBBAs are an invaluable system for ranking the clinical readiness of novel engineered AAV neurotropic capsids, previously selected in animal models or in vitro systems.

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Iannielli, A., Giannelli, S. G., Behrens, T., Luoni, M., Muggeo, S., Moscato, G., Greco, M., Lenzi, E., El Khoury, R., Criscuolo, E., Casucci, M., Clementi, N., Canals, J. M., Iorio, R., Broccoli, V.. 2025-12-23. Fully Human Stem Cell-Derived BBB Model for Clinical Disease Modeling and Neurotropic AAV selection. https://doi.org/10.64898/2025.12.21.695793

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