TY - JOUR
T1 - Attenuation of Heparan Sulfate Proteoglycan Binding Enhances In Vivo Transduction of Human Primary Hepatocytes with AAV2
AU - Cabanes-Creus, Marti
AU - Westhaus, Adrian
AU - Navarro, Renina Gale
AU - Baltazar, Grober
AU - Zhu, Erhua
AU - Amaya, Anais K.
AU - Liao, Sophia H.Y.
AU - Scott, Suzanne
AU - Sallard, Erwan
AU - Dilworth, Kimberley L.
AU - Rybicki, Arkadiusz
AU - Drouyer, Matthieu
AU - Hallwirth, Claus V.
AU - Bennett, Antonette
AU - Santilli, Giorgia
AU - Thrasher, Adrian J.
AU - Agbandje-McKenna, Mavis
AU - Alexander, Ian E.
AU - Lisowski, Leszek
N1 - Publisher Copyright:
© 2020 The Author(s)
PY - 2020/6/12
Y1 - 2020/6/12
N2 - 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. Adeno-associated viral vectors based on serotype 2 transduce primary human hepatocytes in a xenograft model of human liver with low efficiency. Lisowski and colleagues demonstrated that attenuation of the interaction of AAV2 with its primary receptor, heparan sulfate proteoglycan, significantly enhances transduction of human hepatocytes in this preclinical model.
AB - 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. Adeno-associated viral vectors based on serotype 2 transduce primary human hepatocytes in a xenograft model of human liver with low efficiency. Lisowski and colleagues demonstrated that attenuation of the interaction of AAV2 with its primary receptor, heparan sulfate proteoglycan, significantly enhances transduction of human hepatocytes in this preclinical model.
UR - http://www.scopus.com/inward/record.url?scp=85085511681&partnerID=8YFLogxK
U2 - 10.1016/j.omtm.2020.05.004
DO - 10.1016/j.omtm.2020.05.004
M3 - Article
AN - SCOPUS:85085511681
SN - 2329-0501
VL - 17
SP - 1139
EP - 1154
JO - Molecular Therapy - Methods and Clinical Development
JF - Molecular Therapy - Methods and Clinical Development
ER -