title: Recombinant AAV Integration Is Not Associated With Hepatic Genotoxicity in Nonhuman Primates and Patients.
abstract: Recombinant adeno-associated viral vectors (rAAV) currently constitute a real therapeutic strategy for the sustained correction of diverse genetic conditions. Though a wealth of preclinical and clinical studies have been conducted with rAAV, the oncogenic potential of these vectors is still controversial, particularly when considering liver-directed gene therapy. Few preclinical studies and the recent discovery of incomplete wild-type AAV2 genomes integrated in human hepatocellular carcinoma biopsies have raised concerns on rAAV safety. In the present study, we have characterized the integration of both complete and partial rAAV2/5 genomes in nonhuman primate tissues and clinical liver biopsies from a trial aimed to treat acute intermittent porphyria. We applied a new multiplex linear amplification-mediated polymerase chain reaction (PCR) assay capable of detecting integration events that are originated throughout the rAAV genome. The integration rate was low both in nonhuman primates and patient's samples. Importantly, no integration clusters or events were found in genes previously reported to link rAAV integration with hepatocellular carcinoma development, thus showing the absence of genotoxicity of a systemically administered rAAV2/5 in a large animal model and in the clinical context.
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg, Germany.
authors.forename: Irene
authors.initials: I
authors.lastname: Gil-Farina
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg, Germany.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Fronza
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg, Germany; Current address: Division of Applied Genomics, Eurofins Medigenomix GmbH, Ebersberg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Kaeppel
authors.affiliation: Department of Gene Therapy and Hepatology, Center for Applied Medical Research, Pamplona, Spain.
authors.forename: Esperanza
authors.initials: E
authors.lastname: Lopez-Franco
authors.affiliation: Research and Development, uniQure B.V., Amsterdam, The Netherlands.
authors.forename: Valerie
authors.initials: V
authors.lastname: Ferreira
authors.affiliation: Hepatology Unit, Clinica Universidad de Navarra, Pamplona, Spain; Instituto de Investigación Sanitaria de Navarra, Pamplona, Spain.
authors.forename: Delia
authors.initials: D
authors.lastname: D'Avola
authors.affiliation: Hepatology Unit, Clinica Universidad de Navarra, Pamplona, Spain; Instituto de Investigación Sanitaria de Navarra, Pamplona, Spain.
authors.forename: Alberto
authors.initials: A
authors.lastname: Benito
authors.affiliation: Hepatology Unit, Clinica Universidad de Navarra, Pamplona, Spain; Instituto de Investigación Sanitaria de Navarra, Pamplona, Spain.
authors.forename: Jesus
authors.initials: J
authors.lastname: Prieto
authors.affiliation: Research and Development, uniQure B.V., Amsterdam, The Netherlands.
authors.forename: Harald
authors.initials: H
authors.lastname: Petry
authors.affiliation: Department of Gene Therapy and Hepatology, Center for Applied Medical Research, Pamplona, Spain; Instituto de Investigación Sanitaria de Navarra, Pamplona, Spain.
authors.forename: Gloria
authors.initials: G
authors.lastname: Gonzalez-Aseguinolaza
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg, Germany. Electronic address: manfred.schmidt@nct-heidelberg.de.
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics / Q000502:physiology*; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000009:adverse effects; D006801:Humans; D008099:Liver / Q000187:drug effects*; D008252:Macaca fascicularis; D017118:Porphyria, Acute Intermittent / Q000628:therapy*; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA / Q000379:methods; D014161:Transduction, Genetic; D016662:Virus Integration