Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2
Use of the prototypical adeno-associated virus type 2 (AAV2) capsid delivered unexpectedly modest efficacy in an early liver-targeted gene therapy trial for hemophilia B. This result is consistent with subsequent data generated in chimeric mouse-human livers showing that the AAV2 capsid transduces p...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050120300917 |
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doaj-192ac1b2202f420f9e1d2c4d0646e77d |
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Article |
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DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Marti Cabanes-Creus Adrian Westhaus Renina Gale Navarro Grober Baltazar Erhua Zhu Anais K. Amaya Sophia H.Y. Liao Suzanne Scott Erwan Sallard Kimberley L. Dilworth Arkadiusz Rybicki Matthieu Drouyer Claus V. Hallwirth Antonette Bennett Giorgia Santilli Adrian J. Thrasher Mavis Agbandje-McKenna Ian E. Alexander Leszek Lisowski |
| spellingShingle |
Marti Cabanes-Creus Adrian Westhaus Renina Gale Navarro Grober Baltazar Erhua Zhu Anais K. Amaya Sophia H.Y. Liao Suzanne Scott Erwan Sallard Kimberley L. Dilworth Arkadiusz Rybicki Matthieu Drouyer Claus V. Hallwirth Antonette Bennett Giorgia Santilli Adrian J. Thrasher Mavis Agbandje-McKenna Ian E. Alexander Leszek Lisowski Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2 Molecular Therapy: Methods & Clinical Development |
| author_facet |
Marti Cabanes-Creus Adrian Westhaus Renina Gale Navarro Grober Baltazar Erhua Zhu Anais K. Amaya Sophia H.Y. Liao Suzanne Scott Erwan Sallard Kimberley L. Dilworth Arkadiusz Rybicki Matthieu Drouyer Claus V. Hallwirth Antonette Bennett Giorgia Santilli Adrian J. Thrasher Mavis Agbandje-McKenna Ian E. Alexander Leszek Lisowski |
| author_sort |
Marti Cabanes-Creus |
| title |
Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2 |
| title_short |
Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2 |
| title_full |
Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2 |
| title_fullStr |
Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2 |
| title_full_unstemmed |
Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2 |
| title_sort |
attenuation of heparan sulfate proteoglycan binding enhances in vivo transduction of human primary hepatocytes with aav2 |
| publisher |
Elsevier |
| series |
Molecular Therapy: Methods & Clinical Development |
| issn |
2329-0501 |
| publishDate |
2020-06-01 |
| description |
Use of the prototypical adeno-associated virus type 2 (AAV2) capsid delivered unexpectedly modest efficacy in an early liver-targeted gene therapy trial for hemophilia B. This result is consistent with subsequent data generated in chimeric mouse-human livers showing that the AAV2 capsid transduces primary human hepatocytes in vivo with low efficiency. In contrast, novel variants generated by directed evolution in the same model, such as AAV-NP59, transduce primary human hepatocytes with high efficiency. While these empirical data have immense translational implications, the mechanisms underpinning this enhanced AAV capsid transduction performance in primary human hepatocytes are yet to be fully elucidated. Remarkably, AAV-NP59 differs from the prototypical AAV2 capsid by only 11 aa and can serve as a tool to study the correlation between capsid sequence/structure and vector function. Using two orthogonal vectorological approaches, we have determined that just 2 of the 11 changes present in AAV-NP59 (T503A and N596D) account for the enhanced transduction performance of this capsid variant in primary human hepatocytes in vivo, an effect that we have associated with attenuation of heparan sulfate proteoglycan (HSPG) binding affinity. In support of this hypothesis, we have identified, using directed evolution, two additional single amino acid substitution AAV2 variants, N496D and N582S, which are highly functional in vivo. Both substitution mutations reduce AAV2’s affinity for HSPG. Finally, we have modulated the ability of AAV8, a highly murine-hepatotropic serotype, to interact with HSPG. The results support our hypothesis that enhanced HSPG binding can negatively affect the in vivo function of otherwise strongly hepatotropic variants and that modulation of the interaction with HSPG is critical to ensure maximum efficiency in vivo. The insights gained through this study can have powerful implications for studies into AAV biology and capsid development for preclinical and clinical applications targeting liver and other organs. |
| url |
http://www.sciencedirect.com/science/article/pii/S2329050120300917 |
| work_keys_str_mv |
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doaj-192ac1b2202f420f9e1d2c4d0646e77d2020-11-25T03:22:09ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012020-06-011711391154Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2Marti Cabanes-Creus0Adrian Westhaus1Renina Gale Navarro2Grober Baltazar3Erhua Zhu4Anais K. Amaya5Sophia H.Y. Liao6Suzanne Scott7Erwan Sallard8Kimberley L. Dilworth9Arkadiusz Rybicki10Matthieu Drouyer11Claus V. Hallwirth12Antonette Bennett13Giorgia Santilli14Adrian J. Thrasher15Mavis Agbandje-McKenna16Ian E. Alexander17Leszek Lisowski18Translational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, Australia; Great Ormond Street Institute of Child Health, University College London, London WC1N 1EH, UKTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, Australia; Gene Therapy Research Unit, Children’s Medical Research Institute & The Children’s Hospital at Westmead, University of Sydney, Westmead, NSW 2145, AustraliaGene Therapy Research Unit, Children’s Medical Research Institute & The Children’s Hospital at Westmead, University of Sydney, Westmead, NSW 2145, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, AustraliaGene Therapy Research Unit, Children’s Medical Research Institute & The Children’s Hospital at Westmead, University of Sydney, Westmead, NSW 2145, Australia; Commonwealth Scientific and Industrial Research Organisation (CSIRO), North Ryde, NSW 2113, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, AustraliaGene Therapy Research Unit, Children’s Medical Research Institute & The Children’s Hospital at Westmead, University of Sydney, Westmead, NSW 2145, AustraliaDepartment of Biochemistry and Molecular Biology, Center for Structural Biology, University of Florida, Gainesville, FL 32610, USAGreat Ormond Street Institute of Child Health, University College London, London WC1N 1EH, UKGreat Ormond Street Institute of Child Health, University College London, London WC1N 1EH, UKDepartment of Biochemistry and Molecular Biology, Center for Structural Biology, University of Florida, Gainesville, FL 32610, USAGene Therapy Research Unit, Children’s Medical Research Institute & The Children’s Hospital at Westmead, University of Sydney, Westmead, NSW 2145, Australia; Discipline of Child and Adolescent Health, The University of Sydney, Sydney, NSW 2006, AustraliaTranslational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, Australia; Vector and Genome Engineering Facility, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, Australia; Military Institute of Hygiene and Epidemiology, Biological Threats Identification and Countermeasure Center, 24-100 Puławy, Poland; Corresponding author: Leszek Lisowski, Translational Vectorology Research Unit, Children’s Medical Research Institute, The University of Sydney, Westmead, NSW 2145, Australia.Use of the prototypical adeno-associated virus type 2 (AAV2) capsid delivered unexpectedly modest efficacy in an early liver-targeted gene therapy trial for hemophilia B. This result is consistent with subsequent data generated in chimeric mouse-human livers showing that the AAV2 capsid transduces primary human hepatocytes in vivo with low efficiency. In contrast, novel variants generated by directed evolution in the same model, such as AAV-NP59, transduce primary human hepatocytes with high efficiency. While these empirical data have immense translational implications, the mechanisms underpinning this enhanced AAV capsid transduction performance in primary human hepatocytes are yet to be fully elucidated. Remarkably, AAV-NP59 differs from the prototypical AAV2 capsid by only 11 aa and can serve as a tool to study the correlation between capsid sequence/structure and vector function. Using two orthogonal vectorological approaches, we have determined that just 2 of the 11 changes present in AAV-NP59 (T503A and N596D) account for the enhanced transduction performance of this capsid variant in primary human hepatocytes in vivo, an effect that we have associated with attenuation of heparan sulfate proteoglycan (HSPG) binding affinity. In support of this hypothesis, we have identified, using directed evolution, two additional single amino acid substitution AAV2 variants, N496D and N582S, which are highly functional in vivo. Both substitution mutations reduce AAV2’s affinity for HSPG. Finally, we have modulated the ability of AAV8, a highly murine-hepatotropic serotype, to interact with HSPG. The results support our hypothesis that enhanced HSPG binding can negatively affect the in vivo function of otherwise strongly hepatotropic variants and that modulation of the interaction with HSPG is critical to ensure maximum efficiency in vivo. The insights gained through this study can have powerful implications for studies into AAV biology and capsid development for preclinical and clinical applications targeting liver and other organs.http://www.sciencedirect.com/science/article/pii/S2329050120300917 |