title: High-Density Recombinant Adeno-Associated Viral Particles are Competent Vectors for In Vivo Transduction.
abstract: Recombinant adeno-associated viral (rAAV) vectors have recently achieved clinical successes in human gene therapy. However, the commonly observed, heavier particles found in rAAV preparations have traditionally been ignored due to their reported low in vitro transduction efficiency. In this study, the biological properties of regular and high-density rAAV serotype 8 vectors, rAAVRDand rAAVHD, were systemically compared. Results demonstrated that both rAAVRDand rAAVHDexhibited similar DNA packaging profiles, while rAAVHDcapsids contained fewer VP1 and VP2 proteins, indicating that the rAAVHDparticles contained a higher DNA/protein ratio than that of rAAVRDparticles. Dynamic light scattering and transmission electron microscopy data revealed that the diameter of rAAVHDwas smaller than that of rAAVRD. In vitro, rAAVHDwas two- to fourfold less efficient in transduction compared with rAAVRD. However, the transduction performance of rAAVHDand rAAVRDwas similar in vivo. No significant difference in neutralizing antibody formation against rAAVRDand rAAVHDwas observed, suggesting that the surface epitopes of rAAVRDand rAAVHDare congruent. In summary, the results of this study demonstrate that rAAVRDand rAAVHDare equally competent for in vivo transduction, despite their difference in vitro. Therefore, the use of rAAVHDvectors in human gene therapy should be further evaluated.
authors.affiliation: 1 Institute of Genomics, School of Biomedical Sciences, Huaqiao University , Quanzhou, China.|2 Sol Sherry Thrombosis Research Center, Temple University , Philadelphia, Pennsylvania.
authors.forename: Qizhao
authors.initials: Q
authors.lastname: Wang
authors.affiliation: 3 Department of Microbiology and Immunology, Temple University , Philadelphia, Pennsylvania.|4 United States Department of Agriculture, ARS , ERRC, Wyndmoor, Pennyslvania.
authors.forename: Jenni
authors.initials: J
authors.lastname: Firrman
authors.affiliation: 2 Sol Sherry Thrombosis Research Center, Temple University , Philadelphia, Pennsylvania.
authors.forename: Zhongren
authors.initials: Z
authors.lastname: Wu
authors.affiliation: 3 Department of Microbiology and Immunology, Temple University , Philadelphia, Pennsylvania.
authors.forename: Katie A
authors.initials: KA
authors.lastname: Pokiniewski
authors.affiliation: 5 Division of Human Genetics, Cincinnati Children's Hospital Medical Center , Cincinnati, Ohio.|6 Department of Pediatrics, University of Cincinnati School of Medicine , Cincinnati, Ohio.
authors.forename: C Alexander
authors.initials: CA
authors.lastname: Valencia
authors.affiliation: 7 Department of Chemistry, Temple University , Philadelphia, Pennsylvania.
authors.forename: Hairong
authors.initials: H
authors.lastname: Wang
authors.affiliation: 2 Sol Sherry Thrombosis Research Center, Temple University , Philadelphia, Pennsylvania.
authors.forename: Hongying
authors.initials: H
authors.lastname: Wei
authors.affiliation: 1 Institute of Genomics, School of Biomedical Sciences, Huaqiao University , Quanzhou, China.|2 Sol Sherry Thrombosis Research Center, Temple University , Philadelphia, Pennsylvania.
authors.forename: Zhenjing
authors.initials: Z
authors.lastname: Zhuang
authors.affiliation: 4 United States Department of Agriculture, ARS , ERRC, Wyndmoor, Pennyslvania.
authors.forename: LinShu
authors.initials: L
authors.lastname: Liu
authors.affiliation: 7 Department of Chemistry, Temple University , Philadelphia, Pennsylvania.
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Wunder
authors.affiliation: 1 Institute of Genomics, School of Biomedical Sciences, Huaqiao University , Quanzhou, China.
authors.forename: Mario P S
authors.initials: MP
authors.lastname: Chin
authors.affiliation: 1 Institute of Genomics, School of Biomedical Sciences, Huaqiao University , Quanzhou, China.
authors.forename: Ruian
authors.initials: R
authors.lastname: Xu
authors.affiliation: 1 Institute of Genomics, School of Biomedical Sciences, Huaqiao University , Quanzhou, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Diao
authors.affiliation: 2 Sol Sherry Thrombosis Research Center, Temple University , Philadelphia, Pennsylvania.
authors.forename: Biao
authors.initials: B
authors.lastname: Dong
authors.affiliation: 1 Institute of Genomics, School of Biomedical Sciences, Huaqiao University , Quanzhou, China.|2 Sol Sherry Thrombosis Research Center, Temple University , Philadelphia, Pennsylvania.|3 Department of Microbiology and Immunology, Temple University , Philadelphia, Pennsylvania.|4 United States Department of Agriculture, ARS , ERRC, Wyndmoor, Pennyslvania.|8 Cardiovascular Research Center, Temple University , Philadelphia, Pennsylvania.
authors.forename: Weidong
authors.initials: W
authors.lastname: Xiao
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics*; D000229:Dependovirus / Q000235:genetics*; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage*; D006467:Hemophilia A / Q000235:genetics / Q000628:therapy*; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: D036022:Capsid Proteins; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: AAV; capsid; density; gene therapy