Abstract

Adeno-associated virus (AAV) has enjoyed enormous success as a delivery modality for gene therapy yet remains hindered by certain drawbacks. I thus developed two new approaches for circumventing drawbacks inherent in AAV gene therapy. (1) VaultAAV: the AAV delivery modality suffers from high prevalence of preexisting neutralizing antibodies in human populations, limiting who can receive potentially life-saving treatments. As a novel solution to this issue, I employed SpyTag-SpyCatcher molecular glue technology to facilitate packaging of AAVs inside of recombinant protein vault nanoparticles. Vaults are endogenous particles produced by mammalian cells. I therefore hypothesized that they may shield packaged molecules from neutralizing antibodies. Vaults have previously been utilized to deliver drugs and proteins into cells, but my research represents the first time anyone has packaged an entire virus inside of a vault. I showed that the vaultAAV delivery vehicle transduces cells in the presence of anti-AAV neutralizing serum. VaultAAV is positioned as a new gene therapy delivery platform with potential to overcome the neutralizing antibody problem. (2) AdAAV: The small 4.7 kb packaging capacity of the AAV sharply limits the scope of its application. In addition, high doses of AAV are frequently required to facilitate therapeutic effects, leading to acute toxicity issues. To address these challenges, I developed a novel delivery system consisting of adenovirus (Ad) covalently linked to multiple AAV capsids as a new way of more efficiently co-infecting cells with lower overall amounts of AAVs. I utilized the DogTag-DogCatcher (DgT-DgC) molecular glue system to construct AdAAVs and demonstrated that these hybrid virus complexes achieved enhanced co-transduction of cultured cells, including physiologically relevant human primary cells. On this basis, AdAAV technology may eventually facilitate therapeutic co-delivery of multiple transgenes at low virus doses for treating complex ailments.

Committee Chair

David Curiel

Committee Members

Daved Fremont; Farshid Guilak; Jai Rudra; Michael Vahey

Degree

Doctor of Philosophy (PhD)

Author's Department

Biomedical Engineering

Author's School

McKelvey School of Engineering

Document Type

Dissertation

Date of Award

6-29-2026

Language

English (en)

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Available for download on Monday, June 26, 2028

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