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Gene transfer into human hematopoietic progenitor cells with an episomal vector carrying an S/MAR element

Gene Therapy volume 13, pages 40–51 (2006)Cite this article

Abstract

Episomally maintained self-replicating systems present attractive alternative vehicles for gene therapy applications. Recent insights into the ability of chromosomal scaffold/matrix attachment regions (S/MARs) to mediate episomal maintenance of genetic elements allowed the development of a small circular episomal vector that functions independently of virally encoded proteins. In this study, we investigated the potential of this vector, pEPI-eGFP, to mediate gene transfer in hematopoietic progenitor cell lines and primary human cells. pEPI-eGFP was episomally maintained and conferred sustained eGFP expression even in nonselective conditions in the human cell line, K562, as well as in primary human fibroblast-like cells. In contrast, in the murine erythroleukemia cell line, MEL, transgene expression was silenced through histone deacetylation, despite the vector's episomal persistence. Hematopoietic semisolid cell colonies derived from transfected human cord blood CD34+ cells expressed eGFP, albeit at low levels. After 4 weeks, the vector is retained in approximately 1% of progeny cells. Our results provide the first evidence that S/MAR-based plasmids can function as stable episomes in primary human cells, supporting long-term transgene expression. However, they do not display universal behavior in all cell types.

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Acknowledgements

We thank Ioannis Vervitas (Department of Obstetrics, Patras University Hospital) for cord blood collection, Professor Hans Lipps (Institute of Cell Biology, University of Witten, Germany) and Aris Giannakopoulos (Department of Biology, Faculty of Medicine, University of Patras) for providing pEPI-eGFP and pCEP4-eGFP plasmids, respectively, and Zoi Lygerou (Department of Biology, Faculty of Medicine, University of Patras) for her generous gifts of antibodies and fruitful discussions. This work was supported by grants ‘Karatheodori 2003’ (B112, University of Patras) and EPAN 2003 (SP-YB90, EU via GSRT) to AA.

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  1. Department of Biology, Faculty of Medicine, University Hospital of Patras, Rion, Patras, Greece

    E P Papapetrou & A Athanassiadou

  2. Hematology Division, Department of Internal Medicine, University Hospital of Patras, Rion, Patras, Greece

    E P Papapetrou, P G Ziros, I D Micheva & N C Zoumbos

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Correspondence to E P Papapetrou.

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Papapetrou, E., Ziros, P., Micheva, I. et al. Gene transfer into human hematopoietic progenitor cells with an episomal vector carrying an S/MAR element. Gene Ther 13, 40–51 (2006). https://doi.org/10.1038/sj.gt.3302593

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