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Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach
Adeno-associated viruses (AAV) are composed of nonenveloped, icosahedral protein shells that can be adapted to package and deliver recombinant therapeutic DNA. Approaches to engineer recombinant capsids for gene therapy applications have focused on rational design or library-based approaches that ca...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7495376/ https://www.ncbi.nlm.nih.gov/pubmed/32669336 http://dx.doi.org/10.1128/JVI.00976-20 |
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author | Havlik, L. Patrick Simon, Katherine E. Smith, J. Kennon Klinc, Kelli A. Tse, Longping V. Oh, Daniel K. Fanous, Marco M. Meganck, Rita M. Mietzsch, Mario Kleinschmidt, Jürgen Agbandje-McKenna, Mavis Asokan, Aravind |
author_facet | Havlik, L. Patrick Simon, Katherine E. Smith, J. Kennon Klinc, Kelli A. Tse, Longping V. Oh, Daniel K. Fanous, Marco M. Meganck, Rita M. Mietzsch, Mario Kleinschmidt, Jürgen Agbandje-McKenna, Mavis Asokan, Aravind |
author_sort | Havlik, L. Patrick |
collection | PubMed |
description | Adeno-associated viruses (AAV) are composed of nonenveloped, icosahedral protein shells that can be adapted to package and deliver recombinant therapeutic DNA. Approaches to engineer recombinant capsids for gene therapy applications have focused on rational design or library-based approaches that can address one or two desirable attributes; however, there is an unmet need to comprehensively improve AAV vector properties. Such cannot be achieved by utilizing sequence data alone but requires harnessing the three-dimensional (3D) structural properties of AAV capsids. Here, we solve the structures of a natural AAV isolate complexed with antibodies using cryo-electron microscopy and harness this structural information to engineer AAV capsid libraries through saturation mutagenesis of different antigenic footprints. Each surface loop was evolved by infectious cycling in the presence of a helper adenovirus to yield a new AAV variant that then serves as a template for evolving the next surface loop. This stepwise process yielded a humanized AAV8 capsid (AAVhum.8) displaying nonnatural surface loops that simultaneously display tropism for human hepatocytes, increased gene transfer efficiency, and neutralizing antibody evasion. Specifically, AAVhum.8 can better evade neutralizing antisera from multiple species than AAV8. Further, AAVhum.8 displays robust transduction in a human liver xenograft mouse model with expanded tropism for both murine and human hepatocytes. This work supports the hypothesis that critical properties, such as AAV capsid antibody evasion and tropism, can be coevolved by combining rational design and library-based evolution for clinical gene therapy. IMPORTANCE Clinical gene therapy with recombinant AAV vectors has largely relied on natural capsid isolates. There is an unmet need to comprehensively improve AAV tissue tropism, transduction efficiency, and antibody evasion. Such cannot be achieved by utilizing capsid sequence data alone but requires harnessing the 3D structural properties of AAV capsids. Here, we combine rational design and library-based evolution to coevolve multiple, desirable properties onto AAV by harnessing 3D structural information. |
format | Online Article Text |
id | pubmed-7495376 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-74953762020-10-01 Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach Havlik, L. Patrick Simon, Katherine E. Smith, J. Kennon Klinc, Kelli A. Tse, Longping V. Oh, Daniel K. Fanous, Marco M. Meganck, Rita M. Mietzsch, Mario Kleinschmidt, Jürgen Agbandje-McKenna, Mavis Asokan, Aravind J Virol Gene Delivery Adeno-associated viruses (AAV) are composed of nonenveloped, icosahedral protein shells that can be adapted to package and deliver recombinant therapeutic DNA. Approaches to engineer recombinant capsids for gene therapy applications have focused on rational design or library-based approaches that can address one or two desirable attributes; however, there is an unmet need to comprehensively improve AAV vector properties. Such cannot be achieved by utilizing sequence data alone but requires harnessing the three-dimensional (3D) structural properties of AAV capsids. Here, we solve the structures of a natural AAV isolate complexed with antibodies using cryo-electron microscopy and harness this structural information to engineer AAV capsid libraries through saturation mutagenesis of different antigenic footprints. Each surface loop was evolved by infectious cycling in the presence of a helper adenovirus to yield a new AAV variant that then serves as a template for evolving the next surface loop. This stepwise process yielded a humanized AAV8 capsid (AAVhum.8) displaying nonnatural surface loops that simultaneously display tropism for human hepatocytes, increased gene transfer efficiency, and neutralizing antibody evasion. Specifically, AAVhum.8 can better evade neutralizing antisera from multiple species than AAV8. Further, AAVhum.8 displays robust transduction in a human liver xenograft mouse model with expanded tropism for both murine and human hepatocytes. This work supports the hypothesis that critical properties, such as AAV capsid antibody evasion and tropism, can be coevolved by combining rational design and library-based evolution for clinical gene therapy. IMPORTANCE Clinical gene therapy with recombinant AAV vectors has largely relied on natural capsid isolates. There is an unmet need to comprehensively improve AAV tissue tropism, transduction efficiency, and antibody evasion. Such cannot be achieved by utilizing capsid sequence data alone but requires harnessing the 3D structural properties of AAV capsids. Here, we combine rational design and library-based evolution to coevolve multiple, desirable properties onto AAV by harnessing 3D structural information. American Society for Microbiology 2020-09-15 /pmc/articles/PMC7495376/ /pubmed/32669336 http://dx.doi.org/10.1128/JVI.00976-20 Text en Copyright © 2020 Havlik et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Gene Delivery Havlik, L. Patrick Simon, Katherine E. Smith, J. Kennon Klinc, Kelli A. Tse, Longping V. Oh, Daniel K. Fanous, Marco M. Meganck, Rita M. Mietzsch, Mario Kleinschmidt, Jürgen Agbandje-McKenna, Mavis Asokan, Aravind Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach |
title | Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach |
title_full | Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach |
title_fullStr | Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach |
title_full_unstemmed | Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach |
title_short | Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach |
title_sort | coevolution of adeno-associated virus capsid antigenicity and tropism through a structure-guided approach |
topic | Gene Delivery |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7495376/ https://www.ncbi.nlm.nih.gov/pubmed/32669336 http://dx.doi.org/10.1128/JVI.00976-20 |
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