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- Title
Ex Vivo Expansion of Retrovirally Transduced Primate CD34<sup>+</sup> Cells Results in Overrepresentation of Clones With MDS1/EVI1 Insertion Sites in the Myeloid Lineage After Transplantation.
- Authors
Sellers, Stephanie; Gomes, Theotonius J.; Larochelle, Andre; Lopez, Rebecca; Adler, Rima; Krouse, Allen; Donahue, Robert E.; Childs, Richard W.; Dunbar, Cynthia E.
- Abstract
Activation of proto-oncogenes by retroviral insertion is an important issue delaying clinical development of gene therapy. We have reported the nonrandom persistence of hematopoietic clones with vector insertions within the MDS1/EVI1 locus following transplantation of rhesus macaques. We now ask whether prolonged culture of transduced CD34+ cells before transplantation selects for clones with insertions in the MDS1/EVI11 or other proto-oncogene loci. CD34+ cells were transduced with standard retroviral vectors for 4 days and then continued in culture for an additional 6 days before transplantation. A 15% of insertions identified in granulocytes 6 months post-transplant were in MDS1/EVI11, significantly increased compared to the frequency in animals transplanted with cells immediately following transduction. MDS1/EVI1 clones became more dominant over time post-transplantation in one animal that was followed long term, accompanied by an increased overall copy number of vector-containing granulocytes, with one MDS1/EVI1 clone eventually accounting for 100% of transduced granulocytes and marrow colony-forming unit (CFU). This vector insertion increased the expression of Evi1 mRNA. There was no overrepresentation of MDS1/EVI1 insertions contributing to lymphoid lineages. Strategies involving prolonged ex vivo expansion of transduced cells may increase the risk of genotoxicity.
- Subjects
RETROVIRUS diseases; CELL transplantation; HEMATOPOIETIC stem cells; MOLECULAR genetics; MOLECULAR cloning; GENE expression; GENETIC transduction; GENE therapy
- Publication
Molecular Therapy, 2010, Vol 18, Issue 9, p1633
- ISSN
1525-0016
- Publication type
Article
- DOI
10.1038/mt.2010.117