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  • Cell-Based Therapy
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Cell-Based Therapy

Retroviral vector design studies toward hematopoietic stem cell gene therapy for mucopolysaccharidosis type I

Gene Therapy volume 7, pages 1875–1883 (2000)Cite this article

Abstract

To optimize a gene transfer system for hematopoietic stem cell gene therapy of patients with mucopolysaccharidosis (MPS) type I, 10 retroviral vectors were constructed to express the human α-L-iduronidase (IDUA) cDNA. These vectors were designed to evaluate the potential effects of specific promoters, the addition of selectable markers, and the use of multiple promoters versus an internal ribosome entry site for expression of IDUA and selectable maker genes. The effect of vector design was investigated in primary patient fibroblasts (FMPS) or murine fibroblast cell lines; while overall comparison of transgene expression was determined in patients’ peripheral blood lymphocytes (PBLMPS) and CD34+ progenitors (PBPCMPS). We observed that the human PGK promoter introduced the highest IDUA activity per 1% relative transgene frequency in FMPS. Use of the same promoter to separately regulate both the therapeutic gene and a drug-resistance gene resulted in decreased expression of the unselected gene. Co-selection using bicistronic vectors not only increased the number of transductants, but also elevated transgene expression under selective pressure in transgene-positive progenitors. Bicistronic vector LP1CD overcame down-regulation and practically introduced the highest IDUA level in unselected PBLMPS and an intermediate level in PBPCMPS. These studies provide a better understanding of factors contributing to efficient gene expression in hematopoietic cells.

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Acknowledgements

This work was supported by a grant from the National Institute of Child Health and Human Development (PO1-HD32652).

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  1. Department of Pediatrics, Gene Therapy Program, and Institute of Human Genetics, University of Minnesota, Minneapolis, MN, USA

    D Pan, E Aronovich, R S McIvor & C B Whitley

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  1. D Pan
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Pan, D., Aronovich, E., McIvor, R. et al. Retroviral vector design studies toward hematopoietic stem cell gene therapy for mucopolysaccharidosis type I. Gene Ther 7, 1875–1883 (2000). https://doi.org/10.1038/sj.gt.3301298

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