Gene therapy with brain-derived neurotrophic factor as a protection: Retinal ganglion cells in a rat glaucoma model

Keith R G Martin, Harry A Quigley, Donald J Zack, Hana Levkovitch-Verbin, Jennifer Kielczewski, Danielle Valenta, Lisa Baumrind, Mary Ellen Pease, Ronald L. Klein, William W. Hauswirth

Research output: Contribution to journalArticle

Abstract

PURPOSE. To develop a modified adenoassociated viral (AAV) vector capable of efficient transfection of retinal ganglion cells (RGCs) and to test the hypothesis that use of this vector to express brain-derived neurotrophic factor (BDNF) could be protective in experimental glaucoma. METHODS. Ninety-three rats received one unilateral, intravitreal injection of either normal saline (n = 30), AAV-BDNF-wood-chuck hepatitis posttranscriptional regulatory element (WPRE; n = 30), or AAV-green fluorescent protein (GFP)-WPRE (n = 33). Two weeks later, experimental glaucoma was induced in the injected eye by laser application to the trabecular mesh-work. Survival of RGCs was estimated by counting axons in optic nerve cross sections after 4 weeks of glaucoma. Transgene expression was assessed by immunohistochemistry, Western blot analysis, and direct visualization of GFP. RESULTS. The density of GFP-positive cells in retinal whole-mounts was 1,828 ± 299 cells/mm2 (72,273 ± 11,814 cells/retina). Exposure to elevated intraocular pressure was similar in all groups. Four weeks after initial laser treatment, axon loss was 52.3% ± 27.1% in the saline-treated group (n = 25) and 52.3% ± 24.2% in the AAV-GFP-WPRE group (n = 30), but only 32.3% ± 23.0% in the AAV-BDNF-WPRE group (n = 27). Survival in AAV-BDNF-WPRE animals increased markedly and the difference was significant compared with those receiving either AAV-GFP-WPRE (P = 0.002, t-test) or saline (P = 0.006, t-test). CONCLUSIONS. Overexpression of the BDNF gene protects RGC as estimated by axon counts in a rat glaucoma model, further supporting the potential feasibility of neurotrophic therapy as a complement to the lowering of IOP in the treatment of glaucoma.

Original languageEnglish (US)
Pages (from-to)4357-4365
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume44
Issue number10
DOIs
StatePublished - Oct 1 2003

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Retinal Ganglion Cells
Brain-Derived Neurotrophic Factor
Green Fluorescent Proteins
Genetic Therapy
Glaucoma
Axons
Lasers
Intravitreal Injections
Optic Nerve
Intraocular Pressure
Transgenes
Hepatitis
Transfection
Retina
Therapeutics
Western Blotting
Immunohistochemistry
Genes

ASJC Scopus subject areas

  • Ophthalmology

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Gene therapy with brain-derived neurotrophic factor as a protection : Retinal ganglion cells in a rat glaucoma model. / Martin, Keith R G; Quigley, Harry A; Zack, Donald J; Levkovitch-Verbin, Hana; Kielczewski, Jennifer; Valenta, Danielle; Baumrind, Lisa; Pease, Mary Ellen; Klein, Ronald L.; Hauswirth, William W.

In: Investigative Ophthalmology and Visual Science, Vol. 44, No. 10, 01.10.2003, p. 4357-4365.

Research output: Contribution to journalArticle

Martin, Keith R G ; Quigley, Harry A ; Zack, Donald J ; Levkovitch-Verbin, Hana ; Kielczewski, Jennifer ; Valenta, Danielle ; Baumrind, Lisa ; Pease, Mary Ellen ; Klein, Ronald L. ; Hauswirth, William W. / Gene therapy with brain-derived neurotrophic factor as a protection : Retinal ganglion cells in a rat glaucoma model. In: Investigative Ophthalmology and Visual Science. 2003 ; Vol. 44, No. 10. pp. 4357-4365.
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abstract = "PURPOSE. To develop a modified adenoassociated viral (AAV) vector capable of efficient transfection of retinal ganglion cells (RGCs) and to test the hypothesis that use of this vector to express brain-derived neurotrophic factor (BDNF) could be protective in experimental glaucoma. METHODS. Ninety-three rats received one unilateral, intravitreal injection of either normal saline (n = 30), AAV-BDNF-wood-chuck hepatitis posttranscriptional regulatory element (WPRE; n = 30), or AAV-green fluorescent protein (GFP)-WPRE (n = 33). Two weeks later, experimental glaucoma was induced in the injected eye by laser application to the trabecular mesh-work. Survival of RGCs was estimated by counting axons in optic nerve cross sections after 4 weeks of glaucoma. Transgene expression was assessed by immunohistochemistry, Western blot analysis, and direct visualization of GFP. RESULTS. The density of GFP-positive cells in retinal whole-mounts was 1,828 ± 299 cells/mm2 (72,273 ± 11,814 cells/retina). Exposure to elevated intraocular pressure was similar in all groups. Four weeks after initial laser treatment, axon loss was 52.3{\%} ± 27.1{\%} in the saline-treated group (n = 25) and 52.3{\%} ± 24.2{\%} in the AAV-GFP-WPRE group (n = 30), but only 32.3{\%} ± 23.0{\%} in the AAV-BDNF-WPRE group (n = 27). Survival in AAV-BDNF-WPRE animals increased markedly and the difference was significant compared with those receiving either AAV-GFP-WPRE (P = 0.002, t-test) or saline (P = 0.006, t-test). CONCLUSIONS. Overexpression of the BDNF gene protects RGC as estimated by axon counts in a rat glaucoma model, further supporting the potential feasibility of neurotrophic therapy as a complement to the lowering of IOP in the treatment of glaucoma.",
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T2 - Retinal ganglion cells in a rat glaucoma model

AU - Martin, Keith R G

AU - Quigley, Harry A

AU - Zack, Donald J

AU - Levkovitch-Verbin, Hana

AU - Kielczewski, Jennifer

AU - Valenta, Danielle

AU - Baumrind, Lisa

AU - Pease, Mary Ellen

AU - Klein, Ronald L.

AU - Hauswirth, William W.

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N2 - PURPOSE. To develop a modified adenoassociated viral (AAV) vector capable of efficient transfection of retinal ganglion cells (RGCs) and to test the hypothesis that use of this vector to express brain-derived neurotrophic factor (BDNF) could be protective in experimental glaucoma. METHODS. Ninety-three rats received one unilateral, intravitreal injection of either normal saline (n = 30), AAV-BDNF-wood-chuck hepatitis posttranscriptional regulatory element (WPRE; n = 30), or AAV-green fluorescent protein (GFP)-WPRE (n = 33). Two weeks later, experimental glaucoma was induced in the injected eye by laser application to the trabecular mesh-work. Survival of RGCs was estimated by counting axons in optic nerve cross sections after 4 weeks of glaucoma. Transgene expression was assessed by immunohistochemistry, Western blot analysis, and direct visualization of GFP. RESULTS. The density of GFP-positive cells in retinal whole-mounts was 1,828 ± 299 cells/mm2 (72,273 ± 11,814 cells/retina). Exposure to elevated intraocular pressure was similar in all groups. Four weeks after initial laser treatment, axon loss was 52.3% ± 27.1% in the saline-treated group (n = 25) and 52.3% ± 24.2% in the AAV-GFP-WPRE group (n = 30), but only 32.3% ± 23.0% in the AAV-BDNF-WPRE group (n = 27). Survival in AAV-BDNF-WPRE animals increased markedly and the difference was significant compared with those receiving either AAV-GFP-WPRE (P = 0.002, t-test) or saline (P = 0.006, t-test). CONCLUSIONS. Overexpression of the BDNF gene protects RGC as estimated by axon counts in a rat glaucoma model, further supporting the potential feasibility of neurotrophic therapy as a complement to the lowering of IOP in the treatment of glaucoma.

AB - PURPOSE. To develop a modified adenoassociated viral (AAV) vector capable of efficient transfection of retinal ganglion cells (RGCs) and to test the hypothesis that use of this vector to express brain-derived neurotrophic factor (BDNF) could be protective in experimental glaucoma. METHODS. Ninety-three rats received one unilateral, intravitreal injection of either normal saline (n = 30), AAV-BDNF-wood-chuck hepatitis posttranscriptional regulatory element (WPRE; n = 30), or AAV-green fluorescent protein (GFP)-WPRE (n = 33). Two weeks later, experimental glaucoma was induced in the injected eye by laser application to the trabecular mesh-work. Survival of RGCs was estimated by counting axons in optic nerve cross sections after 4 weeks of glaucoma. Transgene expression was assessed by immunohistochemistry, Western blot analysis, and direct visualization of GFP. RESULTS. The density of GFP-positive cells in retinal whole-mounts was 1,828 ± 299 cells/mm2 (72,273 ± 11,814 cells/retina). Exposure to elevated intraocular pressure was similar in all groups. Four weeks after initial laser treatment, axon loss was 52.3% ± 27.1% in the saline-treated group (n = 25) and 52.3% ± 24.2% in the AAV-GFP-WPRE group (n = 30), but only 32.3% ± 23.0% in the AAV-BDNF-WPRE group (n = 27). Survival in AAV-BDNF-WPRE animals increased markedly and the difference was significant compared with those receiving either AAV-GFP-WPRE (P = 0.002, t-test) or saline (P = 0.006, t-test). CONCLUSIONS. Overexpression of the BDNF gene protects RGC as estimated by axon counts in a rat glaucoma model, further supporting the potential feasibility of neurotrophic therapy as a complement to the lowering of IOP in the treatment of glaucoma.

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