title: High-throughput screening and biophysical interrogation of hepatotropic AAV.
abstract: We set out to analyze the fundamental biological differences between AAV2 and AAV8 that may contribute to their different performances in vivo. High-throughput protein interaction screens were used to identify binding partners for each serotype. Of the >8,000 proteins probed, 115 and 134 proteins were identified that interact with AAV2 and AAV8, respectively. Notably, 76 of these protein interactions were shared between the two serotypes. CDK2/cyclinA kinase was identified as a binding partner for both serotypes in the screen. Subsequent analysis confirmed direct binding of CDK2/cyclinA by AAV2 and AAV8. Inhibition of CDK2/cyclinA resulted in increased levels of vector transduction. Biophysical study of vector particle stability and genome uncoating demonstrated slightly greater thermostability for AAV8 than for AAV2. Heat-induced genome uncoating occurred at the same temperature as particle degradation, suggesting that these two processes may be intrinsically related for adeno-associated virus (AAV). Together, these analyses provide insight into commonalities and divergences in the biology of functionally distinct hepatotropic AAV serotypes.
authors.affiliation: Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Samuel L
authors.initials: SL
authors.lastname: Murphy
authors.forename: Anand
authors.initials: A
authors.lastname: Bhagwat
authors.forename: Shyrie
authors.initials: S
authors.lastname: Edmonson
authors.forename: Shangzhen
authors.initials: S
authors.lastname: Zhou
authors.forename: Katherine A
authors.initials: KA
authors.lastname: High
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D001703:Biophysics; D036022:Capsid Proteins / Q000276:immunology / Q000378:metabolism; D019925:Cyclin A / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D019925:Cyclin A; D051357:Cyclin-Dependent Kinase 2