title: High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing.
abstract: We have reported that of the 10 commonly used AAV serotype vectors, AAV6 is the most efficient in transducing primary human hematopoietic stem/progenitor cells (HSPCs). However, the transduction efficiency of the wild-type (WT) AAV6 vector varies greatly in HSPCs from different donors. Here we report two distinct strategies to further increase the transduction efficiency in HSPCs from donors that are transduced less efficiently with the WT AAV6 vectors. The first strategy involved modifications of the viral capsid proteins where specific surface-exposed tyrosine (Y) and threonine (T) residues were mutagenized to generate a triple-mutant (Y705 + Y731F + T492V) AAV6 vector. The second strategy involved the use of ex vivo transduction at high cell density. The combined use of these strategies resulted in transduction efficiency exceeding ~90% in HSPCs at significantly reduced vector doses. Our studies have significant implications in the optimal use of capsid-optimized AAV6 vectors in genome editing in HSPCs.
authors.affiliation: Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA.|Powell Gene Therapy Center; University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Chen
authors.initials: C
authors.lastname: Ling
authors.affiliation: CEA/Paris Sud University (UMR-E 007), Institute of Emerging Diseases and Innovative Therapies (iMETI), Fontenay-aux-Roses, France.
authors.forename: Kanit
authors.initials: K
authors.lastname: Bhukhai
authors.affiliation: Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA.|Powell Gene Therapy Center; University of Florida College of Medicine, Gainesville, FL, USA.|Department of Traditional Chinese Medicine, Second Military Medical University, Shanghai, China.
authors.forename: Zifei
authors.initials: Z
authors.lastname: Yin
authors.affiliation: Department of Physiology, Xiang-Ya School of Medicine, Central South University, Changsha, China.
authors.forename: Mengqun
authors.initials: M
authors.lastname: Tan
authors.affiliation: Herman B Well Center for Pediatrics Research, Indiana University School of Medicine, Indianapolis, IN, USA.
authors.forename: Mervin C
authors.initials: MC
authors.lastname: Yoder
authors.affiliation: CEA/Paris Sud University (UMR-E 007), Institute of Emerging Diseases and Innovative Therapies (iMETI), Fontenay-aux-Roses, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Leboulch
authors.affiliation: CEA/Paris Sud University (UMR-E 007), Institute of Emerging Diseases and Innovative Therapies (iMETI), Fontenay-aux-Roses, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Payen
authors.affiliation: Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA.|Powell Gene Therapy Center; University of Florida College of Medicine, Gainesville, FL, USA.|Department of Molecular Genetics &Microbiology; University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Arun
authors.initials: A
authors.lastname: Srivastava
mesh_terms: D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D002460:Cell Line; D000229:Dependovirus / Q000145:classification / Q000235:genetics*; D000072669:Gene Editing; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000378:metabolism* / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D013045:Species Specificity; D014161:Transduction, Genetic
chemical_list: D036022:Capsid Proteins