May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Heat-inducible AAV-mediated Gene Delivery in RPE Cells
N.J. Sund; X. Yang; A. Kuroki; N. Mirza; J. Bennett; A. Auricchio; M.J. Tolentino
Author Affiliations & Notes
  • N.J. Sund
    Ophthalmology, Scheie Eye Institute/FM Kirby Center for Molecular Ophthalmology, Philadelphia, PA, United States
  • X. Yang
    Ophthalmology, Scheie Eye Institute/FM Kirby Center for Molecular Ophthalmology, Philadelphia, PA, United States
  • A. Kuroki
    Ophthalmology, Scheie Eye Institute/FM Kirby Center for Molecular Ophthalmology, Philadelphia, PA, United States
  • N. Mirza
    Ophthalmology, Scheie Eye Institute/FM Kirby Center for Molecular Ophthalmology, Philadelphia, PA, United States
  • J. Bennett
    Ophthalmology, Scheie Eye Institute/FM Kirby Center for Molecular Ophthalmology, Philadelphia, PA, United States
  • A. Auricchio
    Telethon Institute of Genetics and Medicine (TIGEM), Naples, Italy
  • M.J. Tolentino
    Telethon Institute of Genetics and Medicine (TIGEM), Naples, Italy
  • Footnotes
    Commercial Relationships  N.J. Sund, None; X. Yang, None; A. Kuroki, None; N. Mirza, None; J. Bennett, None; A. Auricchio, None; M.J. Tolentino, None.
  • Footnotes
    Support  JDRF, EY12156, Steinbach Fund, RPB, F.M. Kirby Foundation, P30-DK-47757, AHAF
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 454. doi:
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      N.J. Sund, X. Yang, A. Kuroki, N. Mirza, J. Bennett, A. Auricchio, M.J. Tolentino; Heat-inducible AAV-mediated Gene Delivery in RPE Cells . Invest. Ophthalmol. Vis. Sci. 2003;44(13):454.

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Abstract

Abstract: : Purpose: Tightly-controlling gene delivery of therapeutic proteins to the retina will be important in the treatment of retinal pathology. The purpose of this study is to develop a heat-inducible viral delivery system using an adeno-associated virus (AAV) expressing green fluorescent protein (GFP) under the control of the heat shock protein 70 (hsp70) promoter. Methods: To observe gene activation under different temperatures, recombinant AAV-hsp70-GFP was used to infect human retinal pigment epithelial cell line D407. The cells were then heated between 43 and 47 degrees celsius at different time intervals. Reporter GFP expression was evaluated 24 hours later by fluorescence microscopy and quantified using Image Pro Plus software. Results: AAV-hsp70-GFP is highly-induced in RPE cells when heated compared to non-heat shocked controls. We report the optimal temperature and conditions for maximum transgene expression from this recombinant vector, which is comparable to AAV-GFP under the control of a CMV constitutive promoter. Conclusions: The AAV-hsp70 promoter is active in human RPE cells in vitro and is highly inducible under optimal conditions. This study suggests that an AAV-mediated delivery system based on the hsp70 promoter can deliver therapeutic proteins in a space- and time- specific manner. This may prove useful in developing novel therapies for many types of retinal pathologies.

Keywords: gene transfer/gene therapy • retinal pigment epithelium • retina 
© 2003, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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