Soluble forms of EphrinB2 and EphB4 reduce retinal neovascularization in a model of proliferative retinopathy

David O. Zamora, Michael H. Davies, Stephen Planck, James (Jim) Rosenbaum, Michael (Mike) Powers

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

PURPOSE. Ephrin ligands and their Eph receptors are key regulators of endothelial cell (EC) proliferation, migration, adhesion, and repulsion during mammalian vascular development. The hypothesis was that these molecules also play a role in pathologic neovascularization (NV) in the mouse model of oxygen-induced retinopathy. METHODS. C57BL/6 mice at postnatal day (P)7 were exposed to 75% oxygen (O2) for 5 days (until P12) and allowed to recover in room air to induce retinal NV. Retinas from unexposed and hyperoxia-exposed mice between P7 to P24 were analyzed specifically for EphrinB2 and EphB4 transcript expression by RT-PCR. Phospho-Eph (p-Eph) receptor was evaluated during active EC proliferation at P15 and P17 by immunohistology. Some hyperoxia-exposed mice had one eye injected intravitreally with 150 ng/1.5 μL of soluble EphrinB2/Fc or EphB4/Fc chimeras during transition from high O2 to room air (P12) and injected again on P14. Contralateral eyes were injected with human IgG as the control. Preretinal nuclei and retinal blood vessels were quantified at peak disease (PIT). RESULTS. EphrinB2 mRNA was constitutively expressed in the developing retina and was unchanged by hyperoxia. In contrast, EphB4 mRNA expression was modulated during normal retinal development and was altered by hyperoxia. Furthermore, p-Eph was detected in developing preretinal tufts, thus implying that Ephrin/Eph signaling system is active in this experimental model. Intravitreal injection of soluble versions of these molecules significantly reduced pathologic neovascularization. The number of preretinal nuclei in hyperoxia-treated mice was reduced by 66% (P <0.05) in EphrinB2-injected eyes, whereas EphB4 treatment yielded a 69% reduction (P <0.05), compared with control injections. Intraretinal vessel development was not altered by the injections. CONCLUSIONS. These results support the hypothesis that endogenous EphrinB2 and EphB4 are regulators of retinal NV during oxygen-induced retinopathy and may be useful targets for therapeutic intervention.

Original languageEnglish (US)
Pages (from-to)2175-2182
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume46
Issue number6
DOIs
StatePublished - Jun 2005

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Retinal Neovascularization
Hyperoxia
Ephrins
Pathologic Neovascularization
Oxygen
Retina
EphA1 Receptor
Endothelial Cells
Air
Cell Proliferation
Retinal Vessels
Messenger RNA
Intravitreal Injections
Injections
Inbred C57BL Mouse
Cell Movement
Blood Vessels
Theoretical Models
Immunoglobulin G
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Ophthalmology

Cite this

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title = "Soluble forms of EphrinB2 and EphB4 reduce retinal neovascularization in a model of proliferative retinopathy",
abstract = "PURPOSE. Ephrin ligands and their Eph receptors are key regulators of endothelial cell (EC) proliferation, migration, adhesion, and repulsion during mammalian vascular development. The hypothesis was that these molecules also play a role in pathologic neovascularization (NV) in the mouse model of oxygen-induced retinopathy. METHODS. C57BL/6 mice at postnatal day (P)7 were exposed to 75{\%} oxygen (O2) for 5 days (until P12) and allowed to recover in room air to induce retinal NV. Retinas from unexposed and hyperoxia-exposed mice between P7 to P24 were analyzed specifically for EphrinB2 and EphB4 transcript expression by RT-PCR. Phospho-Eph (p-Eph) receptor was evaluated during active EC proliferation at P15 and P17 by immunohistology. Some hyperoxia-exposed mice had one eye injected intravitreally with 150 ng/1.5 μL of soluble EphrinB2/Fc or EphB4/Fc chimeras during transition from high O2 to room air (P12) and injected again on P14. Contralateral eyes were injected with human IgG as the control. Preretinal nuclei and retinal blood vessels were quantified at peak disease (PIT). RESULTS. EphrinB2 mRNA was constitutively expressed in the developing retina and was unchanged by hyperoxia. In contrast, EphB4 mRNA expression was modulated during normal retinal development and was altered by hyperoxia. Furthermore, p-Eph was detected in developing preretinal tufts, thus implying that Ephrin/Eph signaling system is active in this experimental model. Intravitreal injection of soluble versions of these molecules significantly reduced pathologic neovascularization. The number of preretinal nuclei in hyperoxia-treated mice was reduced by 66{\%} (P <0.05) in EphrinB2-injected eyes, whereas EphB4 treatment yielded a 69{\%} reduction (P <0.05), compared with control injections. Intraretinal vessel development was not altered by the injections. CONCLUSIONS. These results support the hypothesis that endogenous EphrinB2 and EphB4 are regulators of retinal NV during oxygen-induced retinopathy and may be useful targets for therapeutic intervention.",
author = "Zamora, {David O.} and Davies, {Michael H.} and Stephen Planck and Rosenbaum, {James (Jim)} and Powers, {Michael (Mike)}",
year = "2005",
month = "6",
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language = "English (US)",
volume = "46",
pages = "2175--2182",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "6",

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TY - JOUR

T1 - Soluble forms of EphrinB2 and EphB4 reduce retinal neovascularization in a model of proliferative retinopathy

AU - Zamora, David O.

AU - Davies, Michael H.

AU - Planck, Stephen

AU - Rosenbaum, James (Jim)

AU - Powers, Michael (Mike)

PY - 2005/6

Y1 - 2005/6

N2 - PURPOSE. Ephrin ligands and their Eph receptors are key regulators of endothelial cell (EC) proliferation, migration, adhesion, and repulsion during mammalian vascular development. The hypothesis was that these molecules also play a role in pathologic neovascularization (NV) in the mouse model of oxygen-induced retinopathy. METHODS. C57BL/6 mice at postnatal day (P)7 were exposed to 75% oxygen (O2) for 5 days (until P12) and allowed to recover in room air to induce retinal NV. Retinas from unexposed and hyperoxia-exposed mice between P7 to P24 were analyzed specifically for EphrinB2 and EphB4 transcript expression by RT-PCR. Phospho-Eph (p-Eph) receptor was evaluated during active EC proliferation at P15 and P17 by immunohistology. Some hyperoxia-exposed mice had one eye injected intravitreally with 150 ng/1.5 μL of soluble EphrinB2/Fc or EphB4/Fc chimeras during transition from high O2 to room air (P12) and injected again on P14. Contralateral eyes were injected with human IgG as the control. Preretinal nuclei and retinal blood vessels were quantified at peak disease (PIT). RESULTS. EphrinB2 mRNA was constitutively expressed in the developing retina and was unchanged by hyperoxia. In contrast, EphB4 mRNA expression was modulated during normal retinal development and was altered by hyperoxia. Furthermore, p-Eph was detected in developing preretinal tufts, thus implying that Ephrin/Eph signaling system is active in this experimental model. Intravitreal injection of soluble versions of these molecules significantly reduced pathologic neovascularization. The number of preretinal nuclei in hyperoxia-treated mice was reduced by 66% (P <0.05) in EphrinB2-injected eyes, whereas EphB4 treatment yielded a 69% reduction (P <0.05), compared with control injections. Intraretinal vessel development was not altered by the injections. CONCLUSIONS. These results support the hypothesis that endogenous EphrinB2 and EphB4 are regulators of retinal NV during oxygen-induced retinopathy and may be useful targets for therapeutic intervention.

AB - PURPOSE. Ephrin ligands and their Eph receptors are key regulators of endothelial cell (EC) proliferation, migration, adhesion, and repulsion during mammalian vascular development. The hypothesis was that these molecules also play a role in pathologic neovascularization (NV) in the mouse model of oxygen-induced retinopathy. METHODS. C57BL/6 mice at postnatal day (P)7 were exposed to 75% oxygen (O2) for 5 days (until P12) and allowed to recover in room air to induce retinal NV. Retinas from unexposed and hyperoxia-exposed mice between P7 to P24 were analyzed specifically for EphrinB2 and EphB4 transcript expression by RT-PCR. Phospho-Eph (p-Eph) receptor was evaluated during active EC proliferation at P15 and P17 by immunohistology. Some hyperoxia-exposed mice had one eye injected intravitreally with 150 ng/1.5 μL of soluble EphrinB2/Fc or EphB4/Fc chimeras during transition from high O2 to room air (P12) and injected again on P14. Contralateral eyes were injected with human IgG as the control. Preretinal nuclei and retinal blood vessels were quantified at peak disease (PIT). RESULTS. EphrinB2 mRNA was constitutively expressed in the developing retina and was unchanged by hyperoxia. In contrast, EphB4 mRNA expression was modulated during normal retinal development and was altered by hyperoxia. Furthermore, p-Eph was detected in developing preretinal tufts, thus implying that Ephrin/Eph signaling system is active in this experimental model. Intravitreal injection of soluble versions of these molecules significantly reduced pathologic neovascularization. The number of preretinal nuclei in hyperoxia-treated mice was reduced by 66% (P <0.05) in EphrinB2-injected eyes, whereas EphB4 treatment yielded a 69% reduction (P <0.05), compared with control injections. Intraretinal vessel development was not altered by the injections. CONCLUSIONS. These results support the hypothesis that endogenous EphrinB2 and EphB4 are regulators of retinal NV during oxygen-induced retinopathy and may be useful targets for therapeutic intervention.

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