Generation of genetically engineered precursor t-cells from human umbilical cord blood using an optimized alpharetroviral vector platform Journal Article


Authors: Hübner, J.; Hoseini, S. S.; Suerth, J. D.; Hoffmann, D.; Maluski, M.; Herbst, J.; Maul, H.; Ghosh, A.; Eiz-Vesper, B.; Yuan, Q.; Ott, M.; Heuser, M.; Schambach, A.; Sauer, M. G.
Article Title: Generation of genetically engineered precursor t-cells from human umbilical cord blood using an optimized alpharetroviral vector platform
Abstract: Retroviral engineering of hematopoietic stem cell-derived precursor T-cells (preTs) opens the possibility of targeted T-cell transfer across human leukocyte antigen (HLA)-barriers. Alpharetroviral vectors exhibit a more neutral integration pattern thereby reducing the risk of insertional mutagenesis. Cord blood-derived CD34+ cells were transduced and differentiated into preTs in vitro. Two promoters, elongation-factor-1-short-form, and a myeloproliferative sarcoma virus variant in combination with two commonly used envelopes were comparatively assessed choosing enhanced green fluorescent protein or a third-generation chimeric antigen receptor (CAR) against CD123 as gene of interest. Furthermore, the inducible suicide gene iCaspase 9 has been validated. Combining the sarcoma virus-derived promoter with a modified feline endogenous retrovirus envelope glycoprotein yielded in superior transgene expression and transduction rates. Fresh and previously frozen CD34+ cells showed similar transduction and expansion rates. Transgene-positive cells did neither show proliferative impairment nor alteration in their lymphoid differentiation profile. The sarcoma virus-derived promoter only could express sufficient levels of iCaspase 9 to mediate dimerizer-induced apoptosis. Finally, the CD123 CAR was efficiently expressed in CD34+ cells and proved to be functional when expressed on differentiated T-cells. Therefore, the transduction of CD34+ cells with alpharetroviral vectors represents a feasible and potentially safer approach for stem cell-based immunotherapies for cancer. © 2016 The American Society of Gene & Cell Therapy.
Journal Title: Molecular Therapy
Volume: 24
Issue: 7
ISSN: 1525-0016
Publisher: Nature Publishing Group  
Date Published: 2016-07-01
Start Page: 1216
End Page: 1226
Language: English
DOI: 10.1038/mt.2016.89
PROVIDER: scopus
PUBMED: 27138041
PMCID: PMC5088766
DOI/URL:
Notes: Article -- Export Date: 1 September 2016 -- Source: Scopus
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  1. Arnab Ghosh
    65 Ghosh