TY - JOUR
TI - Cell Cycle Status of CD34+Hemopoietic Stem Cells Determines Lentiviral
Integration in Actively Transcribed and Development-related Genes
AU - Papanikolaou, Eleni
AU - Paruzynski, Anna
AU - Kasampalidis, Ioannis and
AU - Deichmann, Annette
AU - Stamateris, Evangelos
AU - Schmidt, Manfred and
AU - von Kalle, Christof
AU - Anagnou, Nicholas P.
JO - Molecular Therapy
PY - 2015
VL - 23
TODO - 4
SP - 683-696
PB - Cell Press
SN - 1525-0016, 1525-0024
TODO - 10.1038/mt.2014.246
TODO - null
TODO - Gene therapy utilizing lentiviral-vectors (LVs) is postulated as a
dynamic therapeutic alternative for monogenic diseases. However,
retroviral gene transfer may cause insertional mutagenesis. Although,
such risks had been originally estimated as extremely low, several
reports of leukemias or clonal dominance, have led to a re-evaluation of
the mechanisms operating in insertional mutagenesis. Therefore,
unraveling the mechanism of retroviral integration is mandatory toward
safer gene therapy applications. In the present study, we undertook an
experimental approach which enabled direct correlation of the cell cycle
stage of the target cell with the integration profile of LVs. CD34+
cells arrested at different stages of cell cycle, were transduced with a
GFP-LV. LAM-PCR was employed for integration site detection, followed by
microarray analysis to correlate transcribed genes with integration
sites. The results indicate that similar to 10% of integration events
occurred in actively transcribed genes and that the cell cycle stage of
target cells affects integration pattern. Specifically, use of thymine
promoted a safer profile, since it significantly reduced integration
within cell cycle-related genes, while we observed increased possibility
for integration into genes related to development, and decreased
possibility for integration within cell cycle and cancer-related genes,
when transduction occurs during mitosis.
ER -