Genotoxic potential of lineage-specific lentivirus vectors carrying the beta-globin locus control region.
Insertional mutagenesis by long terminal repeat (LTR) enhancers in gamma-retrovirus-based vectors (GVs) in clinical trials has prompted deeper investigations into vector genotoxicity. Experimentally, self-inactivating (SIN) lentivirus vectors (LVs) and GV containing internal promoters/enhancers show reduced genotoxicity, although strong ubiquitously-active enhancers dysregulate genes independent of vector type/design. Herein, we explored the genotoxicity of beta-globin (BG) locus control region (LCR), a strong long-range lineage-specific-enhancer, with/without insulator (Ins) elements in LV using primary hematopoietic progenitors to generate in vitro immortalization (IVIM) assay mutants. LCR-containing LV had approximately 200-fold lower transforming potential, compared to the conventional GV. The LCR perturbed expression of few genes in a 300 kilobase (kb) proviral vicinity but no upregulation of genes associated with cancer, including an erythroid-specific transcription factor occurred. A further twofold reduction in transforming activity was observed with insulated LCR-containing LV. Our data indicate that toxicology studies of LCR-containing LV in mice will likely not yield any insertional oncogenesis with the numbers of animals that can be practically studied.
Arumugam
Paritha I
PI
Division of Experimental Hematology/Cancer Biology, Cincinnati Children's Research Foundation, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio 45229, USA.
Higashimoto
Tomoyasu
T
Urbinati
Fabrizia
F
Modlich
Ute
U
Nestheide
Shawna
S
Xia
Ping
P
Fox
Catherine
C
Corsinotti
Andrea
A
Baum
Christopher
C
Malik
Punam
P
D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000009:adverse effects / Q000235:genetics*; D038101:Insulator Elements / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D019901:Locus Control Region / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D016254:Mutagenesis, Insertional; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic; D055544:beta-Globins / Q000235:genetics*
D055544:beta-Globins
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