Publikationen

Molecular Therapy 24 (3): 592-606 (2016-03)

Gogol-Doering A.; Ammar I.; Gupta S.; Bunse M.; Miskey C.; Chen W.; Uckert W.; Schulz T.F.; Izsvak Z.; Ivics Z.

Genome-wide profiling reveals remarkable parallels between insertion site selection properties of the MLV retrovirus and the piggyBac transposon in primary human CD4+ T cells

Description:
The inherent risks associated with vector insertion in gene therapy need to be carefully assessed. We analyzed the genome-wide distributions of Sleeping Beauty (SB) and piggyBac (PB) transposon insertions as well as MLV retrovirus and HIV lentivirus insertions in human CD4+ T cells cells with respect to a panel of 40 chromatin states. The distribution of SB transposon insertions displayed the least deviation from random, while the PB transposon and the MLV retrovirus showed unexpected parallels across all chromatin states. Both MLV and PB insertions are enriched at transcriptional start sites (TSSs) and co-localize with BRD4-associated sites. We demonstrate physical interaction between the PB transposase and BET-domain proteins (including BRD4), suggesting convergent evolution of a tethering mechanism that directs integrating genetic elements into TSSs. We detect unequal biases across the four systems with respect to targeting genes whose deregulation has been previously linked to serious adverse events in gene therapy clinical trials. The SB transposon has the highest theoretical chance of targeting a safe harbor locus in the human genome. The data underscore the significance of vector choice to reduce the mutagenic load on cells in clinical applications.

Keywords: Binding Sites, CD4-Positive T-Lymphocytes, Chromatin, Chromosome Mapping, DNA Transposable Elements, Gene Expression Regulation, Genetic Vectors, Genome-Wide Association Study, Murine Leukemia Virus, Protein Binding, Transcription Initiation Site, Transposases, Virus Integration, Animals

Type: Article

PubMed: 26755332
Official URL: https://doi.org/10.1038/mt.2016.11
MDC Repository: https://edoc.mdc-berlin.de/15375/