Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach
| Autor: | Mario Mietzsch, Aravind Asokan, L. Patrick Havlik, Kelli A. Klinc, Jürgen A. Kleinschmidt, Mavis Agbandje-McKenna, Marco M. Fanous, Daniel K. Oh, J. Kennon Smith, Katherine E. Simon, Rita M. Meganck, Longping V. Tse |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
viruses
Immunology Computational biology adeno-associated virus medicine.disease_cause Antibodies Viral Microbiology law.invention Cell Line 03 medical and health sciences Transduction (genetics) Mice Gene Delivery liver gene delivery 0302 clinical medicine Capsid law Virology medicine Animals Humans neutralizing antibodies Spotlight Neutralizing antibody Adeno-associated virus Tropism 030304 developmental biology 0303 health sciences biology tropism Cryoelectron Microscopy Rational design Genetic Therapy Dependovirus Antibodies Neutralizing gene therapy Molecular Docking Simulation 030220 oncology & carcinogenesis Insect Science biology.protein Recombinant DNA Tissue tropism Hepatocytes Capsid Proteins |
| Zdroj: | Journal of Virology |
| ISSN: | 1098-5514 |
| Popis: | 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. 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|