An anti-VEGF-B antibody fragment induces regression of pre-existing blood vessels in the rat cornea

Yazad Irani, Pierre D. Scotney, Sonja Klebe, Lauren A. Mortimer, Andrew D. Nash, Keryn A. Williams

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

PURPOSE. We tested the ability of an antibody fragment with specificity for vascular endothelial growth factor-B (VEGF-B) to regress nascent and established corneal blood vessels in the rat. METHODS. A single chain variable antibody fragment (scFv) with specificity for VEGF-B was engineered from the 2H10 hybridoma. Binding to rat, mouse, and human VEGF-B was confirmed by surface plasmon resonance. Activity of the anti–VEGF-B scFv on developing and established corneal blood vessels was assessed following unilateral superficial cautery in male and female outbred Sprague Dawley rats. Groups (untreated, control scFv-treated, or anti– VEGF-B scFv-treated) comprised 6 to 22 rats. Treatment consisted of 5 μL scFv, 1 mg/mL, applied topically five times per day for 14 days, or two subconjunctival injections, 50 μg scFv each, applied 7 days apart, or combined topical and subconjunctival treatment. Corneal vessel area was quantified on hematoxylin-stained corneal flat-mounts, and groups were compared using the Mann-Whitney U test, with post hoc Bonferroni correction. Immunohistochemistry for cleaved caspase-3 was performed. RESULTS. Topical anti–VEGF-B scFv therapy alone did not regress corneal blood vessels significantly (P > 0.05). Subconjunctival injection and combined treatment regressed 14-day established corneal blood vessels (25% reduction in vessel area [P = 0.04] and 37% reduction in vessel area [P < 0.001], respectively, compared to results in untreated controls). Cleaved caspase-3 was identified in vascular endothelial cells of anti–VEGF-B scFv-treated corneas. In scFv-treated rats, corneal endothelial cell function was maintained to 12 weeks after treatment and a normal blink reflex was present. CONCLUSIONS. The anti–VEGF-B scFv significantly regressed established but not developing corneal blood vessels in rats.

LanguageEnglish
Pages3404-3413
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume58
Issue number9
DOIs
Publication statusPublished - 1 Jul 2017

Keywords

  • Antibody fragment
  • Corneal neovascularization
  • Eye drops
  • Subconjunctival injection
  • VEGF-B

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Irani, Yazad ; Scotney, Pierre D. ; Klebe, Sonja ; Mortimer, Lauren A. ; Nash, Andrew D. ; Williams, Keryn A. / An anti-VEGF-B antibody fragment induces regression of pre-existing blood vessels in the rat cornea. In: Investigative Ophthalmology and Visual Science. 2017 ; Vol. 58, No. 9. pp. 3404-3413.
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abstract = "PURPOSE. We tested the ability of an antibody fragment with specificity for vascular endothelial growth factor-B (VEGF-B) to regress nascent and established corneal blood vessels in the rat. METHODS. A single chain variable antibody fragment (scFv) with specificity for VEGF-B was engineered from the 2H10 hybridoma. Binding to rat, mouse, and human VEGF-B was confirmed by surface plasmon resonance. Activity of the anti–VEGF-B scFv on developing and established corneal blood vessels was assessed following unilateral superficial cautery in male and female outbred Sprague Dawley rats. Groups (untreated, control scFv-treated, or anti– VEGF-B scFv-treated) comprised 6 to 22 rats. Treatment consisted of 5 μL scFv, 1 mg/mL, applied topically five times per day for 14 days, or two subconjunctival injections, 50 μg scFv each, applied 7 days apart, or combined topical and subconjunctival treatment. Corneal vessel area was quantified on hematoxylin-stained corneal flat-mounts, and groups were compared using the Mann-Whitney U test, with post hoc Bonferroni correction. Immunohistochemistry for cleaved caspase-3 was performed. RESULTS. Topical anti–VEGF-B scFv therapy alone did not regress corneal blood vessels significantly (P > 0.05). Subconjunctival injection and combined treatment regressed 14-day established corneal blood vessels (25{\%} reduction in vessel area [P = 0.04] and 37{\%} reduction in vessel area [P < 0.001], respectively, compared to results in untreated controls). Cleaved caspase-3 was identified in vascular endothelial cells of anti–VEGF-B scFv-treated corneas. In scFv-treated rats, corneal endothelial cell function was maintained to 12 weeks after treatment and a normal blink reflex was present. CONCLUSIONS. The anti–VEGF-B scFv significantly regressed established but not developing corneal blood vessels in rats.",
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An anti-VEGF-B antibody fragment induces regression of pre-existing blood vessels in the rat cornea. / Irani, Yazad; Scotney, Pierre D.; Klebe, Sonja; Mortimer, Lauren A.; Nash, Andrew D.; Williams, Keryn A.

In: Investigative Ophthalmology and Visual Science, Vol. 58, No. 9, 01.07.2017, p. 3404-3413.

Research output: Contribution to journalArticle

TY - JOUR

T1 - An anti-VEGF-B antibody fragment induces regression of pre-existing blood vessels in the rat cornea

AU - Irani, Yazad

AU - Scotney, Pierre D.

AU - Klebe, Sonja

AU - Mortimer, Lauren A.

AU - Nash, Andrew D.

AU - Williams, Keryn A.

PY - 2017/7/1

Y1 - 2017/7/1

N2 - PURPOSE. We tested the ability of an antibody fragment with specificity for vascular endothelial growth factor-B (VEGF-B) to regress nascent and established corneal blood vessels in the rat. METHODS. A single chain variable antibody fragment (scFv) with specificity for VEGF-B was engineered from the 2H10 hybridoma. Binding to rat, mouse, and human VEGF-B was confirmed by surface plasmon resonance. Activity of the anti–VEGF-B scFv on developing and established corneal blood vessels was assessed following unilateral superficial cautery in male and female outbred Sprague Dawley rats. Groups (untreated, control scFv-treated, or anti– VEGF-B scFv-treated) comprised 6 to 22 rats. Treatment consisted of 5 μL scFv, 1 mg/mL, applied topically five times per day for 14 days, or two subconjunctival injections, 50 μg scFv each, applied 7 days apart, or combined topical and subconjunctival treatment. Corneal vessel area was quantified on hematoxylin-stained corneal flat-mounts, and groups were compared using the Mann-Whitney U test, with post hoc Bonferroni correction. Immunohistochemistry for cleaved caspase-3 was performed. RESULTS. Topical anti–VEGF-B scFv therapy alone did not regress corneal blood vessels significantly (P > 0.05). Subconjunctival injection and combined treatment regressed 14-day established corneal blood vessels (25% reduction in vessel area [P = 0.04] and 37% reduction in vessel area [P < 0.001], respectively, compared to results in untreated controls). Cleaved caspase-3 was identified in vascular endothelial cells of anti–VEGF-B scFv-treated corneas. In scFv-treated rats, corneal endothelial cell function was maintained to 12 weeks after treatment and a normal blink reflex was present. CONCLUSIONS. The anti–VEGF-B scFv significantly regressed established but not developing corneal blood vessels in rats.

AB - PURPOSE. We tested the ability of an antibody fragment with specificity for vascular endothelial growth factor-B (VEGF-B) to regress nascent and established corneal blood vessels in the rat. METHODS. A single chain variable antibody fragment (scFv) with specificity for VEGF-B was engineered from the 2H10 hybridoma. Binding to rat, mouse, and human VEGF-B was confirmed by surface plasmon resonance. Activity of the anti–VEGF-B scFv on developing and established corneal blood vessels was assessed following unilateral superficial cautery in male and female outbred Sprague Dawley rats. Groups (untreated, control scFv-treated, or anti– VEGF-B scFv-treated) comprised 6 to 22 rats. Treatment consisted of 5 μL scFv, 1 mg/mL, applied topically five times per day for 14 days, or two subconjunctival injections, 50 μg scFv each, applied 7 days apart, or combined topical and subconjunctival treatment. Corneal vessel area was quantified on hematoxylin-stained corneal flat-mounts, and groups were compared using the Mann-Whitney U test, with post hoc Bonferroni correction. Immunohistochemistry for cleaved caspase-3 was performed. RESULTS. Topical anti–VEGF-B scFv therapy alone did not regress corneal blood vessels significantly (P > 0.05). Subconjunctival injection and combined treatment regressed 14-day established corneal blood vessels (25% reduction in vessel area [P = 0.04] and 37% reduction in vessel area [P < 0.001], respectively, compared to results in untreated controls). Cleaved caspase-3 was identified in vascular endothelial cells of anti–VEGF-B scFv-treated corneas. In scFv-treated rats, corneal endothelial cell function was maintained to 12 weeks after treatment and a normal blink reflex was present. CONCLUSIONS. The anti–VEGF-B scFv significantly regressed established but not developing corneal blood vessels in rats.

KW - Antibody fragment

KW - Corneal neovascularization

KW - Eye drops

KW - Subconjunctival injection

KW - VEGF-B

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U2 - 10.1167/iovs.16-21343

DO - 10.1167/iovs.16-21343

M3 - Article

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JO - Investigative Ophthalmology and Visual Science

T2 - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

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