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Propionate metabolism in cultured human cells after overexpression of recombinant methylmalonyl CoA mutase: Implications for somatic gene therapy

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Somatic Cell and Molecular Genetics

Abstract

Strategies for somatic gene therapy must consider the metabolic consequences of expressing the recombinant gene product in addition to methods for gene transfer and expression. We describe studies of propionate metabolism in cultured cells transfected with methylmalonyl CoA mutase (MCM), the enzyme deficient inmut methylmalonic acidemia. Transfection of MCM intomut fibroblasts restores propionate metabolism to normal levels in a dose-dependent manner. Overexpression of MCM, or the addition of excess propionate, carnitine, or cobalamin, does not increase propionate metabolism in normal human fibroblasts, lymphoblasts, or hepatoma cells, although hepatic cells exhibit >10-fold higher levels of propionate metabolism. Significantly, the restoration of propionate metabolism inmut fibroblasts is disproportionately greater than the efficiency of transfection, suggesting the presence of a cooperative phenomenon between cells. Intercellular participation in propionate metabolism is evident in cocultures of MCM-deficient and propionyl CoA carboxylase-deficient cells. We conclude that the liver is the preferred target for gene therapy of MCM deficiency because of its greater capacity for propionate metabolism and that cooperation between cells could enhance the biological effect of a subpopulation of cells transformed with recombinant MCM.

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Authors and Affiliations

  1. Departments of Cell Biology and Pediatrics, Baylor College of Medicine, 77030, Houston, Texas

    Michael Wilkemeyer, Jozsef Stankovics & Fred D. Ledley

  2. Howard Hughes Medical Institute, USA

    Jozsef Stankovics & Fred D. Ledley

  3. Department of Pediatrics, Baylor College of Medicine, USDA/ARS Children's Nutritional Research Center, Houston, Texas

    Thomas Foy

  4. Department of Pediatrics, University Medical School of Pecs, 7621, Pecs, Hungary

    Jozsef Stankovics

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  1. Michael Wilkemeyer
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Wilkemeyer, M., Stankovics, J., Foy, T. et al. Propionate metabolism in cultured human cells after overexpression of recombinant methylmalonyl CoA mutase: Implications for somatic gene therapy. Somat Cell Mol Genet 18, 493–505 (1992). https://doi.org/10.1007/BF01232646

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  • DOI: https://doi.org/10.1007/BF01232646

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