Pharmacological regulation of protein expression from adeno-associated viral vectors in the eye

Mol Ther. 2002 Aug;6(2):238-42. doi: 10.1006/mthe.2002.0660.

Abstract

The control, over time and space, of the levels of therapeutic proteins is crucial for successful retinal gene therapy. We tested the ability of adeno-associated viral vectors (AAV) delivered intraocularly to release a secreted protein (erythropoietin (Epo) used as a marker) in the eye, either constitutively or in a pharmacologically regulated manner using the dimerizer-inducible transcriptional regulatory system. Following delivery of a constitutively expressing vector to the intravitreal or subretinal space of nude rats, Epo protein was detected in both the anterior chamber and vitreous fluids. A dual-vector system inducible by the dimerizer rapamycin and expressing Epo was administered into the subretinal space in an attempt to achieve pharmacologic control of trangene expression in the eye. Before induction with rapamycin, the intraocular Epo level was negligible. However, following a systemic administration of rapamycin, Epo was detected in the anterior chamber, peaking on day 3 and returning to baseline 2-3 weeks after withdrawal of the drug. Peak-induced Epo in the anterior chamber was proportional to the dose of rapamycin and was not detected in serum. Similar results were obtained following subretinal administration of the vectors in one nonhuman primate. The rapamycin inducible system promises to be useful for developing gene therapies for inherited retinal degeneration and ocular neovascularization.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Dependovirus / genetics*
  • Erythropoietin / genetics
  • Erythropoietin / metabolism
  • Eye / metabolism*
  • Female
  • Genetic Therapy
  • Genetic Vectors*
  • Rats
  • Rats, Nude
  • Rats, Sprague-Dawley
  • Recombinant Proteins

Substances

  • Recombinant Proteins
  • Erythropoietin