Adenovirus-mediated delivery of bcl-2 in conjunction with pde prolongs the delay in photoreceptor cell death in the rd/rd mouse

Academic Article

Abstract

  • Purpose: To test the possibility that adenovirus-mediated co-delivery of the protooncogene bcl-2 with PDE results in an even greater delay the onset of phoioreceptor cell death in the rd/rd mouse than observed after application of PDE alone. Methods- An El, E3-deleted recombinant adenovirus containing the human bcl-2 cDNA (generously provided by D. Merry) driven by the 2.5 kb sequence immediately upstream to the human rhodopsin gene was generated ( Ad.2.5HRPbc/-2). Effects of somatic overexpression of bcl-2 and PDE were assessed after co-injection of 5 × 104 pfu Ad.2.5HRPfec/-2 and 5 × 104 pfu M.CMVPDE into the subretinal space. M.CMVPDE contains the subunit of rod cGMP phosphodiesterase driven by the CMV promoter {Bennett el al, '96 Nature Medicine 2(6):649}. As control, contralateral eyes were injected with 10 pfu Ad.CMVPDE alone. Clinical assessment of retinal degeneration was made with indirect ophthalmoscopy. Expression of bcl-2 and PDE were assessed with RT-PCR and/or. Western analysis and immunohistochemistry. The rate of retinal degeneration was assessed through measurements of widths of photoreceptor layers as a function of time after birth. Results: Expression studies revealed moderate levels of expression of the bcl-2 transgene after injection of Ad.2.5HRPfcc/ 2 but not after injection of M.CMVPDE alone. Histological evaluation revealed that co-injection of bcl-2 with PDE delayed photoreceptor cell death by 7 weeks as compared to 6 weeks after treatment with Ad.CMVPDE alone. Conclusions: Somatic co-delivery of bcl-2 with PDE prolongs the delay in photoreceptor degeneration in the rd/rd mouse. Delivery of bcl-2 and other genes may be useful in conjunction more directed gene-based treatments in limiting the progression of inherited retinal degeneration. Supported by RO1 EY10820-01, RPB, FFB, the Pennsylvania Lions. V & P Mackall Trust and the F. M. Kirby Foundation. None.
  • Authors

    Author List

  • Klatt L; Zeng Y; Bajwa R; Maguire AM; Li Y; Bennett J
  • Volume

  • 38
  • Issue

  • 4