title: Stable integration of recombinant adeno-associated virus vector genomes after transduction of murine hematopoietic stem cells.
abstract: We previously reported that among single-stranded adeno-associated virus (ssAAV) vectors, serotypes 1 through 5, ssAAV1 is the most efficient in transducing murine hematopoietic stem cells (HSCs), but viral second-strand DNA synthesis remains a rate-limiting step. Subsequently, using double-stranded, self-complementary AAV (scAAV) vectors, serotypes 7 through 10, we observed that scAAV7 vectors also transduce murine HSCs efficiently. In the present study, we used scAAV1 and scAAV7 shuttle vectors to transduce HSCs in a murine bone marrow serial transplant model in vivo, which allowed examination of the AAV proviral integration pattern in the mouse genome, as well as recovery and nucleotide sequence analyses of AAV-HSC DNA junction fragments. The proviral genomes were stably integrated, and integration sites were localized to different mouse chromosomes. None of the integration sites was found to be in a transcribed gene, or near a cellular oncogene. None of the animals, monitored for up to 1 year, exhibited pathological abnormalities. Thus, AAV proviral integration-induced risk of oncogenesis was not found in our study, which provides functional confirmation of stable transduction of self-renewing multipotential HSCs by scAAV vectors as well as promise for the use of these vectors in the potential treatment of disorders of the hematopoietic system.
authors.affiliation: Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Zongchao
authors.initials: Z
authors.lastname: Han
authors.forename: Li
authors.initials: L
authors.lastname: Zhong
authors.forename: Njeri
authors.initials: N
authors.lastname: Maina
authors.forename: Zhongbo
authors.initials: Z
authors.lastname: Hu
authors.forename: Xiaomiao
authors.initials: X
authors.lastname: Li
authors.forename: Nitin S
authors.initials: NS
authors.lastname: Chouthai
authors.forename: Daniela
authors.initials: D
authors.lastname: Bischof
authors.forename: Kirsten A
authors.initials: KA
authors.lastname: Weigel-Van Aken
authors.forename: William B
authors.initials: WB
authors.lastname: Slayton
authors.forename: Mervin C
authors.initials: MC
authors.lastname: Yoder
authors.forename: Arun
authors.initials: A
authors.lastname: Srivastava
mesh_terms: D000818:Animals; D001761:Bleomycin; D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D005260:Female; D015316:Genetic Therapy; D005822:Genetic Vectors; D016679:Genome, Viral; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D051379:Mice; D008810:Mice, Inbred C57BL; D011533:Proviruses / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes; D016662:Virus Integration
chemical_list: D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D001761:Bleomycin; C105427:Zeocin