Recombinant AAV vectors are promising candidates for gene therapy of several human genetic disorders. Despite early successes in clinical trials, our understanding of the mechanisms underlying AAV host and tissue tropism is incomplete. During the previous funding period, we made significant progress in mapping novel AAV-glycan receptor interactions and engineering a new class of liver-detargeted AAV vectors. In addition, we optimized structural tools, in vitro cellular assays and in vivo animal studies pertinent to the characterization of AAV vectors. Although this information provided further insight into the biology of certain AAV serotypes, significant gaps exist in our understanding of numerous other AAV strains currently being tested in clinical studies. In the current proposal, we have assembled a team of experienced investigators in the fields of structural virology, glycobiology, vascular biology and AAV vectors to dissect the biology of several naturally occurring as well as new, lab-derived AAV mutants. Specifically, we will map the structural determinants of AAV-glycan interactions using a battery of molecular modeling, computational ligand docking, glycan array and molecular cloning tools. We then propose to reengineer glycan binding footprints on different AAV serotypes to generate novel mutants with altered tissue tropism. Comprehensive studies investigating the role of different tissue glycans as well as integrin co-receptors on the tropism, biodistribution, pharmacokinetics, and transvascular transport of AAV vectors using mouse models will also be undertaken. The long term goal of the proposed studies is to obtain a thorough understanding of AAV biology in different hosts. If successful, the current proposal could provide significant new insight into the influence of glycans and integrins on AAV tissue tropism. In addition to providing a roadmap for structure-driven design of improved AAV vectors, the proposed studies could guide the selection of appropriate AAV strains for further clinical development.
Despite early successes in gene therapy clinical trials, our understanding of viral vectors remains incomplete. Studying the biology of viral vectors using different cross-disciplinary tools is likely to provide new insight into the factors that determine viral tissue tropism in different hosts. If successful, the current proposal could provide a roadmap for designing safer and improved viral vectors for further clinical development.
|Mestre, Humberto; Hablitz, Lauren M; Xavier, Anna Lr et al. (2018) Aquaporin-4-dependent glymphatic solute transport in the rodent brain. Elife 7:|
|Tse, Longping V; Moller-Tank, Sven; Meganck, Rita M et al. (2018) Mapping and Engineering Functional Domains of the Assembly-Activating Protein of Adeno-associated Viruses. J Virol 92:|
|Albright, Blake H; Storey, Claire M; Murlidharan, Giridhar et al. (2018) Mapping the Structural Determinants Required for AAVrh.10 Transport across the Blood-Brain Barrier. Mol Ther 26:510-523|
|Berry, Garrett E; Tse, Longping V (2017) Virus Binding and Internalization Assay for Adeno-associated Virus. Bio Protoc 7:|
|Tse, Longping Victor; Klinc, Kelli A; Madigan, Victoria J et al. (2017) Structure-guided evolution of antigenically distinct adeno-associated virus variants for immune evasion. Proc Natl Acad Sci U S A 114:E4812-E4821|
|Berry, Garrett E; Asokan, Aravind (2016) Chemical Modulation of Endocytic Sorting Augments Adeno-associated Viral Transduction. J Biol Chem 291:939-47|
|Berry, Garrett Edward; Asokan, Aravind (2016) Cellular transduction mechanisms of adeno-associated viral vectors. Curr Opin Virol 21:54-60|
|Tseng, Yu-Shan; Vliet, Kim Van; Rao, Lavanya et al. (2016) Generation and characterization of anti-Adeno-associated virus serotype 8 (AAV8) and anti-AAV9 monoclonal antibodies. J Virol Methods 236:105-110|
|Pierson, Elizabeth E; Keifer, David Z; Asokan, Aravind et al. (2016) Resolving Adeno-Associated Viral Particle Diversity With Charge Detection Mass Spectrometry. Anal Chem 88:6718-25|
|Nelson, Christopher E; Hakim, Chady H; Ousterout, David G et al. (2016) In vivo genome editing improves muscle function in a mouse model of Duchenne muscular dystrophy. Science 351:403-7|
Showing the most recent 10 out of 43 publications