Retroviral vector integration occurs in preferred genomic targets of human bone marrow-repopulating cells.
Increasing use of hematopoietic stem cells for retroviral vector-mediated gene therapy and recent reports on insertional mutagenesis in mice and humans have created intense interest to characterize vector integrations on a genomic level. We studied retrovirally transduced human peripheral blood progenitor cells with bone marrow-repopulating ability in immune-deficient mice. By using a highly sensitive and specific ligation-mediated polymerase chain reaction (PCR) followed by sequencing of vector integration sites, we found a multitude of simultaneously active human stem cell clones 8 weeks after transplantation. Vector integrations occurred with significantly increased frequency into chromosomes 17 and 19 and into specific regions of chromosomes 6, 13, and 16, although most of the chromosomes were targeted. Preferred genomic target sites have previously only been reported for wild-type retroviruses. Our findings reveal for the first time that retroviral vector integration into human marrow-repopulating cells can be nonrandom (P =.000 37).
Laufs
Stephanie
S
German Cancer Research Center, Research Program Diagnostics and Experimental Therapy, Heidelberg, Germany.
Gentner
Bernhard
B
Nagy
K Zsuzsanna
KZ
Jauch
Anna
A
Benner
Axel
A
Naundorf
Sonja
S
Kuehlcke
Klaus
K
Schiedlmeier
Bernhard
B
Ho
Anthony D
AD
Zeller
W Jens
WJ
Fruehauf
Stefan
S
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002882:Chromosomes, Human, Pair 13; D002885:Chromosomes, Human, Pair 16; D002886:Chromosomes, Human, Pair 17; D002888:Chromosomes, Human, Pair 19; D002896:Chromosomes, Human, Pair 6; D003001:Cloning, Molecular; D005260:Female; D005822:Genetic Vectors; D006410:Hematopoiesis; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016133:Polymerase Chain Reaction / Q000379:methods; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection; D016662:Virus Integration