Portable transcranial therapeutic ultrasound enhances targeted gene delivery for Parkinson's disease: from rodent models to non-human primates

Batts, A. J.; Ji, R.; Bae, S.; Tsitsos, F. N.; Jimenez-Gambin, S.; Kwon, N.; Gorman, S. L.; Tsakri, D.; Noel, R. L.; Bendig, J.; Jimenez, D. A.; DiBenedetto, M.; Del Castillo, S. A.; Keleta, F. B.; Caicedo, J.; Romanov, A.; Curley, C. T.; Dzhashiashvili, Y.; Walton-Gibbs, G. D.; Hollidge, B. S.; Przedborski, S.; Ramsburg, E.; Danos, O.; Ferrera, V. P.; Engel, E. A.; Smith, J. B.; Konofagou, E. E.

doi:10.1101/2025.03.02.639893

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bioRxiv · 10.1101/2025.03.02.639893

Portable transcranial therapeutic ultrasound enhances targeted gene delivery for Parkinson's disease: from rodent models to non-human primates

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Abstract

Gene therapy for neurodegenerative diseases faces significant challenges due to the blood-brain barrier (BBB), which limits drug delivery to the central nervous system (CNS). While clinical trials for Parkinsons disease (PD) have progressed, administration of vectors expressing enzymatic or neurotrophic factor transgenes have required extensive optimization of the delivery method to achieve potentially therapeutic levels of transgene expression. Focused ultrasound (FUS) combined with microbubbles has emerged as a promising non-invasive strategy to transiently open the BBB for targeted gene delivery via viral nanocarriers including recombinant adeno-associated viruses (AAVs). However, key factors influencing FUS-mediated AAV delivery, including dose distribution and therapeutic efficacy, remain underexplored in non-human primates (NHPs). Here, we evaluated the feasibility of AAV9-CAG-GFP delivery using two portable therapeutic ultrasound modalities: ultrasound-guided, spherically-focused FUS (USgFUS) and a novel low-frequency linear array configuration for imaging and therapy called theranostic ultrasound (ThUS). In mice, FUS-sonicated regions exhibited a 25-fold increase in AAV9 biodistribution compared to systemic injection alone. Extending this approach to NHPs, we observed up to a 200-fold increase in AAV9 DNA in treated brain regions, including PD-relevant structures. In assessing the translational therapeutic potential of this technique, ThUS-mediated AAV9-hSyn-hNTRN (human neurturin) delivery in a toxin mouse model of PD facilitated the rescue of up to 80% and 75% of degenerated dopaminergic neurons in the substantia nigra and striatum, respectively. These findings demonstrate that portable ultrasound technologies can non-invasively enhance AAV9 delivery to targeted brain regions in both mice and NHPs relative to what can be achieved with intravenous (IV) delivery of the same capsid alone. With further development, these approaches may offer a clinically viable, non-invasive alternative for gene therapy in neurodegenerative diseases. One sentence summaryBBB opening with portable therapeutic ultrasound non-invasively increased viral gene delivery to the brain after systemic AAV vector administration in mice and rhesus macaques.

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Batts, A. J., Ji, R., Bae, S., Tsitsos, F. N., Jimenez-Gambin, S., Kwon, N., Gorman, S. L., Tsakri, D., Noel, R. L., Bendig, J., Jimenez, D. A., DiBenedetto, M., Del Castillo, S. A., Keleta, F. B., Caicedo, J., Romanov, A., Curley, C. T., Dzhashiashvili, Y., Walton-Gibbs, G. D., Hollidge, B. S., Przedborski, S., Ramsburg, E., Danos, O., Ferrera, V. P., Engel, E. A., Smith, J. B., Konofagou, E. E.. 2025-03-03. Portable transcranial therapeutic ultrasound enhances targeted gene delivery for Parkinson's disease: from rodent models to non-human primates. https://doi.org/10.1101/2025.03.02.639893

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