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Article

Adeno-Associated Virus (AAV) Capsid Stability and Liposome Remodeling During Endo/Lysosomal pH Trafficking

1
Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA
2
Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT 59717, USA
3
Department of Chemistry, College of Liberal Arts & Sciences, University of Florida, Gainesville, FL 32611, USA
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Powell Gene Therapy Center Vector Core, University of Florida, Gainesville, FL 32610, USA
5
Department of Ophthalmology, University of Florida, Gainesville, FL 32610, USA
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Institute of Virology, Charité Medical School, 12203 Berlin, Germany
*
Author to whom correspondence should be addressed.
Current address: Bridget Lins-Austin, Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD 20993, USA.
Received: 25 May 2020 / Revised: 13 June 2020 / Accepted: 16 June 2020 / Published: 20 June 2020
(This article belongs to the Special Issue In Memory of Michael Rossmann)
Adeno-associated viruses (AAVs) are small, non-pathogenic ssDNA viruses being used as therapeutic gene delivery vectors for the treatment of a variety of monogenic diseases. An obstacle to successful gene delivery is inefficient capsid trafficking through the endo/lysosomal pathway. This study aimed to characterize the AAV capsid stability and dynamics associated with this process for a select number of AAV serotypes, AAV1, AAV2, AAV5, and AAV8, at pHs representative of the early and late endosome, and the lysosome (6.0, 5.5, and 4.0, respectively). All AAV serotypes displayed thermal melt temperatures that varied with pH. The stability of AAV1, AAV2, and AAV8 increased in response to acidic conditions and then decreased at pH 4.0. In contrast, AAV5 demonstrated a consistent decrease in thermostability in response to acidification. Negative-stain EM visualization of liposomes in the presence of capsids at pH 5.5 or when heat shocked showed induced remodeling consistent with the externalization of the PLA2 domain of VP1u. These observations provide clues to the AAV capsid dynamics that facilitate successful infection. Finally, transduction assays revealed a pH and temperature dependence with low acidity and temperatures > 4 °C as detrimental factors. View Full-Text
Keywords: AAV; trafficking; capsid; liposomes; PLA2; thermostability; infectivity AAV; trafficking; capsid; liposomes; PLA2; thermostability; infectivity
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MDPI and ACS Style

Lins-Austin, B.; Patel, S.; Mietzsch, M.; Brooke, D.; Bennett, A.; Venkatakrishnan, B.; Van Vliet, K.; Smith, A.N.; Long, J.R.; McKenna, R.; Potter, M.; Byrne, B.; Boye, S.L.; Bothner, B.; Heilbronn, R.; Agbandje-McKenna, M. Adeno-Associated Virus (AAV) Capsid Stability and Liposome Remodeling During Endo/Lysosomal pH Trafficking. Viruses 2020, 12, 668. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060668

AMA Style

Lins-Austin B, Patel S, Mietzsch M, Brooke D, Bennett A, Venkatakrishnan B, Van Vliet K, Smith AN, Long JR, McKenna R, Potter M, Byrne B, Boye SL, Bothner B, Heilbronn R, Agbandje-McKenna M. Adeno-Associated Virus (AAV) Capsid Stability and Liposome Remodeling During Endo/Lysosomal pH Trafficking. Viruses. 2020; 12(6):668. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060668

Chicago/Turabian Style

Lins-Austin, Bridget, Saajan Patel, Mario Mietzsch, Dewey Brooke, Antonette Bennett, Balasubramanian Venkatakrishnan, Kim Van Vliet, Adam N. Smith, Joanna R. Long, Robert McKenna, Mark Potter, Barry Byrne, Sanford L. Boye, Brian Bothner, Regine Heilbronn, and Mavis Agbandje-McKenna. 2020. "Adeno-Associated Virus (AAV) Capsid Stability and Liposome Remodeling During Endo/Lysosomal pH Trafficking" Viruses 12, no. 6: 668. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060668

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