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Hypothalamic delivery of doxycycline-inducible leptin gene allows for reversible transgene expression and physiological responses

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Abstract

Our purpose was to incorporate regulation into the recombinant adeno-associated virus encoding leptin by introducing a tet-inducible promotor. This system, TET-Ob, allows for control of leptin gene expression via doxycycline in drinking water. F344XBN rats (aged 4 months) were given a hypothalamic injection of TET-Ob or control virus. During 34 days of doxycycline (doxy) administration to all rats (STAGE 1), TET-Ob rats gained 50.7% less mass, ate 10.4% less food, and had a 77.5% reduction in serum leptin as compared with controls. Doxy was then withdrawn from half of the TET-Ob rats for 32 days (TET-Ob-OFF), while half continued to receive doxy (TET-Ob-ON) (stage 2). During stage 2, TET-Ob-ON rats gained 44.8% less mass than TET-Ob-OFF and ate significantly less food than both TET-Ob-OFF and controls. Serum leptin increased to 83.4% of control values in TET-Ob-OFF, but remained very low in the in TET-Ob-ON. At death, visceral adiposity was 14.5% of controls in TET-Ob-ON animals, but had risen to 76.9% of controls in TET-Ob-OFF. A reversible increase in both leptin signal transduction in the hypothalamus and uncoupling protein expression in brown adipose was recorded. This system allows for more precise regulation of gene therapy-mediated fat loss.

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Acknowledgements

This work was supported by the Medical Research Service of the Department of Veterans Affairs and National Institute on Aging Grant AG-17047.

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Authors and Affiliations

  1. Department of Veterans Affairs Medical Center, Geriatric Research, Education and Clinical Center, Gainesville, FL, USA

    J Wilsey, E W Shek, M K Matheny & P J Scarpace

  2. Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, FL, USA

    J Wilsey, E W Shek, M K Matheny & P J Scarpace

  3. Department of Molecular Genetics, University of Florida College of Medicine, Gainesville, FL, USA

    S Zolotukhin & V Prima

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  1. J Wilsey
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  2. S Zolotukhin
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Wilsey, J., Zolotukhin, S., Prima, V. et al. Hypothalamic delivery of doxycycline-inducible leptin gene allows for reversible transgene expression and physiological responses. Gene Ther 9, 1492–1499 (2002). https://doi.org/10.1038/sj.gt.3301835

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  • DOI: https://doi.org/10.1038/sj.gt.3301835

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