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Retroviral vector-mediated in vivo expression of low-density-lipoprotein receptors in the Watanabe heritable hyperlipidemic rabbit

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Somatic Cell and Molecular Genetics

Abstract

We have achieved in vivo expression of recombinant low-density-lipoprotein (LDL) receptors in the Watanabe heritable hyperlipidemic (WHHL) rabbit, an animal model for the human disease familial hypercholesterolemia. A retroviral vector was constructed containing the human LDL receptor cDNA and was used to stably transduce primary skin fibroblasts from WHHL rabbits. The integrity and function of the introduced LDL receptor was established by immunoprecipitation, by a fluorescent LDL binding assay, and by the ability of the transduced cells to suppress 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase activity in response to exogenous cholesterol. Autologous transduced fibroblasts were reimplanted into donor rabbits; in vivo LDL receptor expression and the survival of the transduced cells were analyzed by immunohistochemistry and by LDL binding assays performed on cells recovered from the implants. LDL receptor-bearing cells could be identified on tissue sections and recovered from implants for up to four weeks. Total and LDL cholesterol levels decreased significantly after implantation of the transduced cells; however, control experiments indicated that the decreases were not mediated through the recombinant LDL receptor. While in vivo stable expression of recombinant LDL receptors in Watanabe rabbits is possible, consequent changes in lipid levels must be interpreted with caution. This system of site-specific in vivo expression of recombinant LDL receptors permits further evaluation of the role of LDL receptor-gene replacement in the therapy of hypercholesterolemia.

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Author notes

  1. James A. Zwiebel

    Present address: Lombardi Cancer Center, Georgetown University, 20007, Washington D.C.

Authors and Affiliations

  1. Molecular Hematology Branch, National Heart, Lung, and Blood Institute, 20892, Bethesda, Maryland

    David A. Dichek, James A. Zwiebel & W. French Anderson

  2. Molecular Diseases Branch, National Heart, Lung, and Blood Institute, 20892, Bethesda, Maryland

    Zafarul H. Beg

  3. Armed Forces Institute of Pathology, 20306, Washington, D.C.

    Gary L. Bratthauer & Jeffrey M. Hoeg

  4. Childrens' National Medical Center, 20010, Washington, D.C.

    Kathryn D. Anderson & Kurt D. Newman

Authors
  1. David A. Dichek
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  2. Gary L. Bratthauer
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  3. Zafarul H. Beg
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  4. Kathryn D. Anderson
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  6. James A. Zwiebel
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Dichek, D.A., Bratthauer, G.L., Beg, Z.H. et al. Retroviral vector-mediated in vivo expression of low-density-lipoprotein receptors in the Watanabe heritable hyperlipidemic rabbit. Somat Cell Mol Genet 17, 287–301 (1991). https://doi.org/10.1007/BF01232823

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  • DOI: https://doi.org/10.1007/BF01232823

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