title: Vector design influences hepatic genotoxicity after adeno-associated virus gene therapy.
abstract: The use of adeno-associated virus (AAV) as a gene therapy vector has been approved recently for clinical use and has demonstrated efficacy in a growing number of clinical trials. However, the safety of AAV as a vector has been challenged by a single study that documented hepatocellular carcinoma (HCC) after AAV gene delivery in mice. Most studies have not noted genotoxicity following AAV-mediated gene delivery; therefore, the possibility that there is an association between AAV and HCC is controversial. Here, we performed a comprehensive study of HCC in a large number of mice following therapeutic AAV gene delivery. Using a sensitive high-throughput integration site-capture technique and global expressional analysis, we found that AAV integration into the RNA imprinted and accumulated in nucleus (Rian) locus, and the resulting overexpression of proximal microRNAs and retrotransposon-like 1 (Rtl1) were associated with HCC. In addition, we demonstrated that the AAV vector dose, enhancer/promoter selection, and the timing of gene delivery are all critical factors for determining HCC incidence after AAV gene delivery. Together, our results define aspects of AAV-mediated gene therapy that influence genotoxicity and suggest that these features should be considered for design of both safer AAV vectors and gene therapy studies.
authors.forename: Randy J
authors.initials: RJ
authors.lastname: Chandler
authors.forename: Matthew C
authors.initials: MC
authors.lastname: LaFave
authors.forename: Gaurav K
authors.initials: GK
authors.lastname: Varshney
authors.forename: Niraj S
authors.initials: NS
authors.lastname: Trivedi
authors.forename: Nuria
authors.initials: N
authors.lastname: Carrillo-Carrasco
authors.forename: Julien S
authors.initials: JS
authors.lastname: Senac
authors.forename: Weiwei
authors.initials: W
authors.lastname: Wu
authors.forename: Victoria
authors.initials: V
authors.lastname: Hoffmann
authors.forename: Abdel G
authors.initials: AG
authors.lastname: Elkahloun
authors.forename: Shawn M
authors.initials: SM
authors.lastname: Burgess
authors.forename: Charles P
authors.initials: CP
authors.lastname: Venditti
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D000229:Dependovirus; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D051379:Mice; D008817:Mice, Mutant Strains; D011257:Pregnancy Proteins / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: D011257:Pregnancy Proteins; C526773:Rtl1 protein, mouse