Adeno-associated virus capsid assembly is divergent and stochastic

Tobias P Wörner; Antonette Bennett; Sana Habka; Joost Snijder; Olga Friese; Thomas Powers; Mavis Agbandje-McKenna; Albert J R Heck

doi:10.1038/s41467-021-21935-5

Adeno-associated virus capsid assembly is divergent and stochastic

Tobias P Wörner, Antonette Bennett, Sana Habka, Joost Snijder, Olga Friese, Thomas Powers, Mavis Agbandje-McKenna, Albert J R Heck

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Adeno-associated viruses (AAVs) are increasingly used as gene therapy vectors. AAVs package their genome in a non-enveloped T = 1 icosahedral capsid of ~3.8 megaDalton, consisting of 60 subunits of 3 distinct viral proteins (VPs), which vary only in their N-terminus. While all three VPs play a role in cell-entry and transduction, their precise stoichiometry and structural organization in the capsid has remained elusive. Here we investigate the composition of several AAV serotypes by high-resolution native mass spectrometry. Our data reveal that the capsids assemble stochastically, leading to a highly heterogeneous population of capsids of variable composition, whereby even the single-most abundant VP stoichiometry represents only a small percentage of the total AAV population. We estimate that virtually every AAV capsid in a particular preparation has a unique composition. The systematic scoring of the simulations against experimental native MS data offers a sensitive new method to characterize these therapeutically important heterogeneous capsids.

Original language	English
Article number	1642
Pages (from-to)	1-9
Number of pages	9
Journal	Nature Communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-21935-5
Publication status	Published - 12 Mar 2021

Bibliographical note

Funding Information:
We thank the members of the Heck laboratory for general support, especially Arjan Barendregt. This research received funding through the Netherlands Organization for Scientific Research (NWO) TTW project 15575 (Structural analysis and position-resolved imaging of macromolecular structures using novel mass spectrometry–based approaches) and the Spinoza Award SPI.2017.028 to A.J.R.H. A.B. and M.A.-M. are supported by NIH R01 GM109524, NSF DMS 1563234, and funds from the UF College of Medicine. J.S. is supported by the Dutch Research Council NWO Gravitation 2013 BOO, Institute for Chemical Immunology (ICI, 024.002.009). Additionally, we are grateful for the support from the Pfizer Biotherapeutics Pharmaceutical Sciences organization.

Publisher Copyright:
© 2021, The Author(s).

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abstract = "Adeno-associated viruses (AAVs) are increasingly used as gene therapy vectors. AAVs package their genome in a non-enveloped T = 1 icosahedral capsid of ~3.8 megaDalton, consisting of 60 subunits of 3 distinct viral proteins (VPs), which vary only in their N-terminus. While all three VPs play a role in cell-entry and transduction, their precise stoichiometry and structural organization in the capsid has remained elusive. Here we investigate the composition of several AAV serotypes by high-resolution native mass spectrometry. Our data reveal that the capsids assemble stochastically, leading to a highly heterogeneous population of capsids of variable composition, whereby even the single-most abundant VP stoichiometry represents only a small percentage of the total AAV population. We estimate that virtually every AAV capsid in a particular preparation has a unique composition. The systematic scoring of the simulations against experimental native MS data offers a sensitive new method to characterize these therapeutically important heterogeneous capsids.",

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AU - Powers, Thomas

AU - Agbandje-McKenna, Mavis

AU - Heck, Albert J R

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Adeno-associated virus capsid assembly is divergent and stochastic

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