Murine leukemia virus vector integration favors promoter regions and regional hot spots in a human T-cell line.
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.
Tsukahara
Tomonori
T
Department of Microbiology and Immunology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan.
Agawa
Hideyuki
H
Matsumoto
Sayori
S
Matsuda
Mizuho
M
Ueno
Shuichi
S
Yamashita
Yuki
Y
Yamada
Koichiro
K
Tanaka
Nobuyuki
N
Kojima
Katsuhiko
K
Takeshita
Toshikazu
T
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
D012333:RNA, Messenger