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

L. Klatt, Y. Zeng, R. Bajwa, A. M. Maguire, Yiwen Li, J. Bennett

Research output: Contribution to journalArticle

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 × 10 4 pfu Ad.2.5HRPfec/-2 and 5 × 10 4 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.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number4
StatePublished - Dec 1 1997
Externally publishedYes

Fingerprint

Photoreceptor Cells
Adenoviridae
Retinal Degeneration
Cell Death
Injections
Lions
bcl-2 Genes
Human Adenoviruses
Ophthalmoscopy
Rhodopsin
Phosphoric Diester Hydrolases
Transgenes
Genes
Complementary DNA
Immunohistochemistry
Medicine
Parturition
Polymerase Chain Reaction
Therapeutics

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Adenovirus-mediated delivery of bcl-2 in conjunction with pde prolongs the delay in photoreceptor cell death in the rd/rd mouse. / Klatt, L.; Zeng, Y.; Bajwa, R.; Maguire, A. M.; Li, Yiwen; Bennett, J.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 4, 01.12.1997.

Research output: Contribution to journalArticle

@article{6410b1b9bf25417592e934e107d47fd9,
title = "Adenovirus-mediated delivery of bcl-2 in conjunction with pde prolongs the delay in photoreceptor cell death in the rd/rd mouse",
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 × 10 4 pfu Ad.2.5HRPfec/-2 and 5 × 10 4 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.",
author = "L. Klatt and Y. Zeng and R. Bajwa and Maguire, {A. M.} and Yiwen Li and J. Bennett",
year = "1997",
month = "12",
day = "1",
language = "English (US)",
volume = "38",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "4",

}

TY - JOUR

T1 - Adenovirus-mediated delivery of bcl-2 in conjunction with pde prolongs the delay in photoreceptor cell death in the rd/rd mouse

AU - Klatt, L.

AU - Zeng, Y.

AU - Bajwa, R.

AU - Maguire, A. M.

AU - Li, Yiwen

AU - Bennett, J.

PY - 1997/12/1

Y1 - 1997/12/1

N2 - 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 × 10 4 pfu Ad.2.5HRPfec/-2 and 5 × 10 4 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.

AB - 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 × 10 4 pfu Ad.2.5HRPfec/-2 and 5 × 10 4 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.

UR - http://www.scopus.com/inward/record.url?scp=33749132825&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749132825&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:33749132825

VL - 38

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 4

ER -