Increased tau expression in the APOE4 blood-brain barrier model is associated with reduced anti-tau therapeutic antibody delivery in vitro

Tau protein is a critical driver of neurodegeneration and an important drug target in Alzheimer’s disease (AD). Tau-specific immunotherapy has emerged as a promising treatment strategy for AD, however the therapeutic efficacy of anti-tau antibodies may be limited by their insufficient delivery across the blood-brain barrier (BBB). The apolipoprotein E4 allele ( APOE4 ) is the strongest genetic risk factor for sporadic AD and is known to influence tau-mediated neurodegeneration. Interestingly, both tau and APOE4 have been implicated in the cerebrovascular pathology observed in AD. Yet, the crosstalk between APOE4 and tau at the level of the BBB and its consequences for anti-tau immunotherapeutics delivery, remain poorly understood. Here, we utilised APOE3 - and APOE4 -carrying human iPSC-derived induced brain endothelial-like cells (iBECs) as a sporadic AD BBB model, determined the levels of endogenous tau in iBECs, and explored the transport of two novel monoclonal anti-tau antibodies, RNF5 and RN2N, across the in vitro barrier. Our results demonstrate that MAPT gene transcription, tau protein levels and tau phosphorylation are increased in iBECs in an APOE4 -related manner and are associated with reduced iBEC monolayer integrity and increased permeability to biologically inert fluorescent tracers. Additionally, elevated levels of intracellular tau in APOE4 cells were accompanied by the reduced passive permeability of therapeutic anti-tau antibodies through the APOE4 iBEC monolayer, which could be improved by the application of focused ultrasound and microbubble drug-delivery technology. Together, our study illustrates a new role for APOE4 and tau in human iBECs with potential implications for BBB dysfunction and anti-tau therapeutic antibody delivery. ### Competing Interest Statement The authors have declared no competing interest. * AD : Alzheimer’s disease APOE : apolipoprotein E BBB : blood-brain barrier BEC : brain endothelial cell CSF : cerebrospinal fluid CNS : central nervous system ELISA : enzyme-linked immunosorbent assay FUS : focused ultrasound hiPSC : human induced pluripotent stem cell iBEC : induced brain endothelial-like cell IgG : Immunoglobulin G MAPT : microtubule-associated protein tau gene MB : microbubble PSEN1 : presenilin-1 p-tau : phosphorylated tau qPCR : quantitative polymerase chain reaction TEER : trans-endothelial electrical resistance