title: The fetal mouse is a sensitive genotoxicity model that exposes lentiviral-associated mutagenesis resulting in liver oncogenesis.
abstract: Genotoxicity models are extremely important to assess retroviral vector biosafety before gene therapy. We have developed an in utero model that demonstrates that hepatocellular carcinoma (HCC) development is restricted to mice receiving nonprimate (np) lentiviral vectors (LV) and does not occur when a primate (p) LV is used regardless of woodchuck post-translation regulatory element (WPRE) mutations to prevent truncated X gene expression. Analysis of 839 npLV and 244 pLV integrations in the liver genomes of vector-treated mice revealed clear differences between vector insertions in gene dense regions and highly expressed genes, suggestive of vector preference for insertion or clonal outgrowth. In npLV-associated clonal tumors, 56% of insertions occurred in oncogenes or genes associated with oncogenesis or tumor suppression and surprisingly, most genes examined (11/12) had reduced expression as compared with control livers and tumors. Two examples of vector-inserted genes were the Park 7 oncogene and Uvrag tumor suppressor gene. Both these genes and their known interactive partners had differential expression profiles. Interactive partners were assigned to networks specific to liver disease and HCC via ingenuity pathway analysis. The fetal mouse model not only exposes the genotoxic potential of vectors intended for gene therapy but can also reveal genes associated with liver oncogenesis.
authors.affiliation: National Centre for Tumorigenesis, Heidelberg Technology park TP4, Heidelberg, Germany.
authors.forename: Ali
authors.initials: A
authors.lastname: Nowrouzi
authors.forename: Wing T
authors.initials: WT
authors.lastname: Cheung
authors.forename: Tingting
authors.initials: T
authors.lastname: Li
authors.forename: Xuegong
authors.initials: X
authors.lastname: Zhang
authors.forename: Anne
authors.initials: A
authors.lastname: Arens
authors.forename: Anna
authors.initials: A
authors.lastname: Paruzynski
authors.forename: Simon N
authors.initials: SN
authors.lastname: Waddington
authors.forename: Emma
authors.initials: E
authors.lastname: Osejindu
authors.forename: Safia
authors.initials: S
authors.lastname: Reja
authors.forename: Christof
authors.initials: C
authors.lastname: von Kalle
authors.forename: Yoahe
authors.initials: Y
authors.lastname: Wang
authors.forename: Faisal
authors.initials: F
authors.lastname: Al-Allaf
authors.forename: Lisa
authors.initials: L
authors.lastname: Gregory
authors.forename: Matthew
authors.initials: M
authors.lastname: Themis
authors.forename: Maxine
authors.initials: M
authors.lastname: Holder
authors.forename: Niraja
authors.initials: N
authors.lastname: Dighe
authors.forename: Alaine
authors.initials: A
authors.lastname: Ruthe
authors.forename: Suzanne Mk
authors.initials: SM
authors.lastname: Buckley
authors.forename: Brian
authors.initials: B
authors.lastname: Bigger
authors.forename: Eugenio
authors.initials: E
authors.lastname: Montini
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
authors.forename: Robert
authors.initials: R
authors.lastname: Andrews
authors.forename: Terry P
authors.initials: TP
authors.lastname: Roberts
authors.forename: Robert F
authors.initials: RF
authors.lastname: Newbold
authors.forename: Charles
authors.initials: C
authors.lastname: Coutelle
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.forename: Mike
authors.initials: M
authors.lastname: Themis
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000628:therapy; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D004249:DNA Damage; D004195:Disease Models, Animal; D005333:Fetus / Q000473:pathology*; D015870:Gene Expression; D020869:Gene Expression Profiling; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000009:adverse effects* / Q000379:methods; D005822:Genetic Vectors; D016678:Genome; D006678:HIV / Q000235:genetics; D007150:Immunohistochemistry; D004860:Infectious Anemia Virus, Equine / Q000235:genetics*; D008099:Liver / Q000473:pathology*; D008113:Liver Neoplasms / Q000473:pathology / Q000628:therapy; D051379:Mice; D016296:Mutagenesis; D016254:Mutagenesis, Insertional; D009154:Mutation; D060888:Real-Time Polymerase Chain Reaction