title: Murine leukemia virus vector integration favors promoter regions and regional hot spots in a human T-cell line.
abstract: Genomic analysis of integration will be important in evaluating the safety of human gene therapy with retroviral vectors. Here, we investigated MLV vector integration sites in human T-cells, since they are amenable to gene transfer studies, and have been used therapeutically in clinical trials. We mapped 340 MLV vector integration sites in the infected human T-cell clones we established. The data showed that MLV preferred integration near the transcription start sites (+/-5kb), near CpG islands (+/-1kb), and within the first intron of RefSeq genes. We also identified MLV integration hot spots that contained three or more integrations within a 100kb region. RT-PCR revealed that mRNA-levels of T-cell clones that contained MLV integrations near transcription start sites or introns were dysregulated compared to the uninfected cells. These studies help define the profile of MLV integration in T-cells and the risks associated with MLV-based gene therapy.
authors.affiliation: Department of Microbiology and Immunology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Tomonori
authors.initials: T
authors.lastname: Tsukahara
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Agawa
authors.forename: Sayori
authors.initials: S
authors.lastname: Matsumoto
authors.forename: Mizuho
authors.initials: M
authors.lastname: Matsuda
authors.forename: Shuichi
authors.initials: S
authors.lastname: Ueno
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yamashita
authors.forename: Koichiro
authors.initials: K
authors.lastname: Yamada
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Tanaka
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Kojima
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Takeshita
mesh_terms: D002460:Cell Line; D003001:Cloning, Molecular; D018899:CpG Islands; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics*; D016296:Mutagenesis; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism* / Q000821:virology; D024363:Transcription Initiation Site; D016662:Virus Integration
chemical_list: D012333:RNA, Messenger