Ultrasound-mediated delivery of novel tau-specific monoclonal antibody enhances brain uptake but not therapeutic efficacy

Bajracharya, Rinie; Cruz, Esteban; Götz, Jürgen; Nisbet, Rebecca M.

doi:10.1016/j.jconrel.2022.07.026

Cited by 16 publications

(31 citation statements)

References 88 publications

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“…The effect of FUS +MB was tested on iBECs 48-72h after subculture on Transwell inserts. FITC-conjugated dextran (150 kDa) was added at 0.5 mg/ml and AlexaFluor™-647-conjugated anti-amyloid-β (Aducanumab-analogue, 27 ) and anti-Tau (RNF5, 36 ) therapeutic antibodies were added at 1 μM. MBs (10 μL per Transwell) were then added to the wells aseptically directly before the FUS treatment.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, we developed a sporadic AD patient iPSC-derived BBB-like in vitro model to study the effects of FUS +MB on cellular responses and drug delivery. We utilized APOE3- and APOE4- carrying iPSC-derived BBB cells and investigated molecular changes following FUS +MB as well as the FUS +MB -mediated delivery of two therapeutic AD antibodies: an Aducanumab analogue (anti-Aβ antibody, commercially known as Aduhelm TM ) 34 , 35 , and RNF5, an anti-Tau antibody, which has recently been shown to be efficient in reducing p-Tau levels in an animal model with Tau pathology 36 . Here, we report the improved delivery of these antibodies in our BBB-like in vitro model using FUS +MB and the cell-specific effects FUS +MB confers in an APOE3 / APOE4 context.…”

Section: Introductionmentioning

confidence: 99%

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A sporadic Alzheimer's blood-brain barrier model for developing ultrasound-mediated delivery of Aducanumab and anti-Tau antibodies

Wasielewska¹,

Chaves²,

Johnston³

et al. 2022

Theranostics

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Rationale: The blood-brain barrier (BBB) is a major impediment to therapeutic intracranial drug delivery for the treatment of neurodegenerative diseases, including Alzheimer's disease (AD). Focused ultrasound applied together with microbubbles (FUS +MB ) is a novel technique to transiently open the BBB and increase drug delivery. Evidence suggests that FUS +MB is safe, however, the effects of FUS +MB on human BBB cells, especially in the context of AD, remain sparsely investigated. In addition, there currently are no cell platforms to test for FUS +MB -mediated drug delivery. Methods: Here we generated BBB cells (induced brain endothelial-like cells (iBECs) and astrocytes (iAstrocytes)) from apolipoprotein E gene allele E4 ( APOE4 , high sporadic AD risk) and allele E3 ( APOE3 , lower AD risk) carrying patient-derived induced pluripotent stem cells (iPSCs). We established mono- and co-culture models of human sporadic AD and control BBB cells to investigate the effects of FUS +MB on BBB cell phenotype and to screen for the delivery of two potentially therapeutic AD antibodies, an Aducanumab-analogue (Aduhelm TM ; anti-amyloid-β) and a novel anti-Tau antibody, RNF5. We then developed a novel hydrogel-based 2.5D BBB model as a step towards a more physiologically relevant FUS +MB drug delivery platform. Results: When compared to untreated cells, the delivery of Aducanumab-analogue and RNF5 was significantly increased (up to 1.73 fold), across the Transwell-based BBB models following FUS +MB treatment. Our results also demonstrated the safety of FUS +MB indicated by minimal changes in iBEC transcriptome as well as little or no changes in iBEC or iAstrocyte viability and inflammatory responses within the first 24 h post FUS +MB . Furthermore, we demonstrated successful iBEC barrier formation in our novel 2.5D hydrogel-based BBB model with significantly increased delivery (1.4 fold) of Aducanumab-analogue following FUS +MB . Conclusion: Our results demonstrate a robust and reproducible approach to utilize patient cells for FUS +MB -mediated drug delivery screening in vitro . With such a cell platform for FUS +MB research previously not reported, it has the potential to identify novel FUS +MB -deliverable drugs as well as screen for cell- and patient-specific effects of FUS +MB , accelerating the use of FUS +MB as a therapeutic modality in AD.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A sporadic Alzheimer's blood-brain barrier model for developing ultrasound-mediated delivery of Aducanumab and anti-Tau antibodies

Wasielewska¹,

Chaves²,

Johnston³

et al. 2022

Theranostics

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“…Hence we hypothesised that increased levels of tau in APOE4 iBECs may contribute to limited anti-antibody passage through the BBB. To investigate this, we further utilised the tau-targeting antibodies RNF5 and RN2N shown to achieve therapeutic effects in tau transgenic mouse models [42][43][44] and screened their permeability in vitro in our Transwell-based APOE BBB model 41 .…”

Section: Passive Permeability Of Anti-tau Therapeutic Antibodies Is R...mentioning

confidence: 99%

“…Having observed reduced passive permeability of anti-tau antibodies in APOE4 cells, we hypothesised that active delivery methods may be required to facilitate the delivery of tautargeting immunotherapeutics in APOE4 iBECs. Focused ultrasound and microbubble (FUS +MB ) technology is an emerging method successfully utilised to enhance large molecule drug delivery in preclinical AD studies performed by us and others 42,44,45,[56][57][58] and proven safe and clinically effective in temporary BBB opening in AD patients 57,[59][60][61][62] . We have previously established an ultrasound-mediated antibody delivery in vitro platform where we demonstrated that FITC-conjugated 150 kDa dextran, AF647-RNF5 and AF647-Aducanumab permeability can be increased in a sporadic AD BBB model by the application of FUS +MB at optimised parameters 41 .…”

Section: Active Delivery With Fus +Mb Enhances the Permeability Of Ta...mentioning

confidence: 99%

Increased tau expression in theAPOE4blood-brain barrier model is associated with reduced anti-tau therapeutic antibody deliveryin vitro

Wasielewska,

Bajracharya,

Johnston

et al. 2023

Preprint

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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 andAPOE4have been implicated in the cerebrovascular pathology observed in AD. Yet, the crosstalk betweenAPOE4and tau at the level of the BBB and its consequences for anti-tau immunotherapeutics delivery, remain poorly understood. Here, we utilisedAPOE3- andAPOE4-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 thein vitrobarrier. Our results demonstrate thatMAPTgene transcription, tau protein levels and tau phosphorylation are increased in iBECs in anAPOE4-related manner and are associated with reduced iBEC monolayer integrity and increased permeability to biologically inert fluorescent tracers. Additionally, elevated levels of intracellular tau inAPOE4cells were accompanied by the reduced passive permeability of therapeutic anti-tau antibodies through theAPOE4iBEC monolayer, which could be improved by the application of focused ultrasound and microbubble drug-delivery technology. Together, our study illustrates a new role forAPOE4and tau in human iBECs with potential implications for BBB dysfunction and anti-tau therapeutic antibody delivery.

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“…These results suggest that the N‐terminus was accessible in fibrillar and non‐fibrillar recombinant Aβ 1‐42 aggregates and consistent with all available in vitro structures of Aβ 1‐42 (Creekmore et al, 2021). Similarly, all anti‐tau N‐terminal monoclonal antibodies TAU1 (tau 1‐22 ), RNF5 (tau 35‐44 ) (Bajracharya et al, 2021), and HJ8.5 (Yanamandra et al, 2013) (tau 25‐30 ) (Table S2) showed tight binding to recombinant hTau441 heparin‐induced filaments (Figures 5b and S8d,e). That was again consistent with the structures of heparin‐induced hTau441 filaments with accessible N‐termini (Zhang et al, 2019).…”

Section: Resultsmentioning

confidence: 81%

Quantitative proteomics of tau and Aβ in detergent fractions from Alzheimer's disease brains

Mukherjee

Dubois

Perez

et al. 2022

Journal of Neurochemistry

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The two hallmarks of Alzheimer's disease (AD) are amyloidβ (Aβ) plaques and neurofibrillary tangles marked by phosphorylated tau. Increasing evidence suggests that aggregating Aβ drives tau accumulation, a process that involves synaptic degeneration leading to cognitive impairment. Conversely, there is a realization that non-fibrillar (oligomeric) forms of Aβ mediate toxicity in AD. Fibrillar (filamentous) aggregates of proteins across the spectrum of the primary and secondary tauopathies were the focus of recent structural studies with a filament structure-based nosologic classification, but less emphasis was given to non-filamentous co-aggregates of insoluble proteins in the fractions derived from post-mortem human brains. Here, we revisited sarkosyl-soluble and -insoluble extracts to characterize tau and Aβ species by quantitative targeted mass spectrometric proteomics, biochemical assays, and electron microscopy. AD brain sarkosyl-insoluble pellets were greatly enriched with Aβ 42 at almost equimolar levels to N-terminal truncated microtubule-binding region (MTBR) isoforms of tau with multiple site-specific post-translational modifications (PTMs). MTBR R3 and R4 tau peptides were most abundant in the sarkosyl-insoluble materials with a 10-fold higher concentration than N-terminal tau peptides. This indicates that the major proportion of the enriched tau was the aggregation-prone N-terminal and proline-rich region (PRR) of truncated mixed 4R and 3R tau with more 4R than 3R isoforms. High concentration and occupancies of site-specific phosphorylation pT 181 (~22%) and pT 217 (~16%) (key biomarkers of AD) along with other PTMs in the PRR and MTBR indicated a regional susceptibility of PTMs in aggregated tau. Immunogold

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Ultrasound-mediated delivery of novel tau-specific monoclonal antibody enhances brain uptake but not therapeutic efficacy

Cited by 16 publications

References 88 publications

A sporadic Alzheimer's blood-brain barrier model for developing ultrasound-mediated delivery of Aducanumab and anti-Tau antibodies

A sporadic Alzheimer's blood-brain barrier model for developing ultrasound-mediated delivery of Aducanumab and anti-Tau antibodies

Increased tau expression in theAPOE4blood-brain barrier model is associated with reduced anti-tau therapeutic antibody deliveryin vitro

Quantitative proteomics of tau and Aβ in detergent fractions from Alzheimer's disease brains

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