Neurofibromin deficiency induces endothelial cell proliferation and retinal neovascularization

Hanfang Zhang, Farlyn Z. Hudson, Zhimin Xu, Rebekah Tritz, Modesto Antonio Rojas, Chintan Patel, Stephen B. Haigh, Zsuzsanna Bordán, David A. Ingram, David J Fulton, Neal Lee Weintraub, Ruth B Caldwell, Brian Kevin Stansfield

Research output: Contribution to journalArticle

Abstract

PURPOSE. Neurofibromatosis type 1 (NF1) is the result of inherited mutations in the NF1 tumor suppressor gene, which encodes the protein neurofibromin. Eye manifestations are common in NF1 with recent reports describing a vascular dysplasia in the retina and choroid. Common features of NF1 retinopathy include tortuous and dilated feeder vessels that terminate in capillary tufts, increased endothelial permeability, and neovascularization. Given the retinal vascular phenotype observed in persons with NF1, we hypothesize that preserving neurofibromin may be a novel strategy to control pathologic retinal neovascularization. METHODS. Nf1 expression in human endothelial cells (EC) was reduced using small hairpin (sh) RNA and EC proliferation, migration, and capacity to form vessel-like networks were assessed in response to VEGF and hypoxia. Wild-type (WT), Nf1 heterozygous (Nf1 +/- ), and Nf1 flox/+ ;Tie2cre pups were subjected to hyperoxia/hypoxia using the oxygen-induced retinopathy model. Retinas were analyzed quantitatively for extent of retinal vessel dropout, neovascularization, and capillary branching. RESULTS. Neurofibromin expression was suppressed in response to VEGF, which corresponded with activation of Mek-Erk and PI3-K-Akt signaling. Neurofibromin-deficient EC exhibited enhanced proliferation and network formation in response to VEGF and hypoxia via an Akt-dependent mechanism. In response to hyperoxia/hypoxia, Nf1 +/- retinas exhibited increased vessel dropout and neovascularization when compared with WT retinas. Neovascularization was similar between Nf1 +/- and Nf1 flox/+ ;Tie2cre retinas, but capillary drop out in Nf1 flox/+ ;Tie2cre retinas was significantly reduced when compared with Nf1 +/- retinas. CONCLUSIONS. These data suggest that neurofibromin expression is essential for controlling endothelial cell proliferation and retinal neovascularization and therapies targeting neurofibromin-deficient EC may be beneficial.

Original languageEnglish (US)
Pages (from-to)2520-2528
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume59
Issue number6
DOIs
StatePublished - May 1 2018

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Neurofibromin 1
Retinal Neovascularization
Retina
Neurofibromatosis 1
Endothelial Cells
Cell Proliferation
Vascular Endothelial Growth Factor A
Retinal Vessels
Hyperoxia
Eye Manifestations
Pathologic Neovascularization
Choroid
Tumor Suppressor Genes
Small Interfering RNA
Cell Movement
Blood Vessels
Permeability
Oxygen
Phenotype
Mutation

Keywords

  • Endothelial cell
  • Neurofibromatosis
  • Ras
  • Retinopathy of prematurity
  • VEGF

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Neurofibromin deficiency induces endothelial cell proliferation and retinal neovascularization. / Zhang, Hanfang; Hudson, Farlyn Z.; Xu, Zhimin; Tritz, Rebekah; Rojas, Modesto Antonio; Patel, Chintan; Haigh, Stephen B.; Bordán, Zsuzsanna; Ingram, David A.; Fulton, David J; Weintraub, Neal Lee; Caldwell, Ruth B; Stansfield, Brian Kevin.

In: Investigative Ophthalmology and Visual Science, Vol. 59, No. 6, 01.05.2018, p. 2520-2528.

Research output: Contribution to journalArticle

Zhang, Hanfang ; Hudson, Farlyn Z. ; Xu, Zhimin ; Tritz, Rebekah ; Rojas, Modesto Antonio ; Patel, Chintan ; Haigh, Stephen B. ; Bordán, Zsuzsanna ; Ingram, David A. ; Fulton, David J ; Weintraub, Neal Lee ; Caldwell, Ruth B ; Stansfield, Brian Kevin. / Neurofibromin deficiency induces endothelial cell proliferation and retinal neovascularization. In: Investigative Ophthalmology and Visual Science. 2018 ; Vol. 59, No. 6. pp. 2520-2528.
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abstract = "PURPOSE. Neurofibromatosis type 1 (NF1) is the result of inherited mutations in the NF1 tumor suppressor gene, which encodes the protein neurofibromin. Eye manifestations are common in NF1 with recent reports describing a vascular dysplasia in the retina and choroid. Common features of NF1 retinopathy include tortuous and dilated feeder vessels that terminate in capillary tufts, increased endothelial permeability, and neovascularization. Given the retinal vascular phenotype observed in persons with NF1, we hypothesize that preserving neurofibromin may be a novel strategy to control pathologic retinal neovascularization. METHODS. Nf1 expression in human endothelial cells (EC) was reduced using small hairpin (sh) RNA and EC proliferation, migration, and capacity to form vessel-like networks were assessed in response to VEGF and hypoxia. Wild-type (WT), Nf1 heterozygous (Nf1 +/- ), and Nf1 flox/+ ;Tie2cre pups were subjected to hyperoxia/hypoxia using the oxygen-induced retinopathy model. Retinas were analyzed quantitatively for extent of retinal vessel dropout, neovascularization, and capillary branching. RESULTS. Neurofibromin expression was suppressed in response to VEGF, which corresponded with activation of Mek-Erk and PI3-K-Akt signaling. Neurofibromin-deficient EC exhibited enhanced proliferation and network formation in response to VEGF and hypoxia via an Akt-dependent mechanism. In response to hyperoxia/hypoxia, Nf1 +/- retinas exhibited increased vessel dropout and neovascularization when compared with WT retinas. Neovascularization was similar between Nf1 +/- and Nf1 flox/+ ;Tie2cre retinas, but capillary drop out in Nf1 flox/+ ;Tie2cre retinas was significantly reduced when compared with Nf1 +/- retinas. CONCLUSIONS. These data suggest that neurofibromin expression is essential for controlling endothelial cell proliferation and retinal neovascularization and therapies targeting neurofibromin-deficient EC may be beneficial.",
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T1 - Neurofibromin deficiency induces endothelial cell proliferation and retinal neovascularization

AU - Zhang, Hanfang

AU - Hudson, Farlyn Z.

AU - Xu, Zhimin

AU - Tritz, Rebekah

AU - Rojas, Modesto Antonio

AU - Patel, Chintan

AU - Haigh, Stephen B.

AU - Bordán, Zsuzsanna

AU - Ingram, David A.

AU - Fulton, David J

AU - Weintraub, Neal Lee

AU - Caldwell, Ruth B

AU - Stansfield, Brian Kevin

PY - 2018/5/1

Y1 - 2018/5/1

N2 - PURPOSE. Neurofibromatosis type 1 (NF1) is the result of inherited mutations in the NF1 tumor suppressor gene, which encodes the protein neurofibromin. Eye manifestations are common in NF1 with recent reports describing a vascular dysplasia in the retina and choroid. Common features of NF1 retinopathy include tortuous and dilated feeder vessels that terminate in capillary tufts, increased endothelial permeability, and neovascularization. Given the retinal vascular phenotype observed in persons with NF1, we hypothesize that preserving neurofibromin may be a novel strategy to control pathologic retinal neovascularization. METHODS. Nf1 expression in human endothelial cells (EC) was reduced using small hairpin (sh) RNA and EC proliferation, migration, and capacity to form vessel-like networks were assessed in response to VEGF and hypoxia. Wild-type (WT), Nf1 heterozygous (Nf1 +/- ), and Nf1 flox/+ ;Tie2cre pups were subjected to hyperoxia/hypoxia using the oxygen-induced retinopathy model. Retinas were analyzed quantitatively for extent of retinal vessel dropout, neovascularization, and capillary branching. RESULTS. Neurofibromin expression was suppressed in response to VEGF, which corresponded with activation of Mek-Erk and PI3-K-Akt signaling. Neurofibromin-deficient EC exhibited enhanced proliferation and network formation in response to VEGF and hypoxia via an Akt-dependent mechanism. In response to hyperoxia/hypoxia, Nf1 +/- retinas exhibited increased vessel dropout and neovascularization when compared with WT retinas. Neovascularization was similar between Nf1 +/- and Nf1 flox/+ ;Tie2cre retinas, but capillary drop out in Nf1 flox/+ ;Tie2cre retinas was significantly reduced when compared with Nf1 +/- retinas. CONCLUSIONS. These data suggest that neurofibromin expression is essential for controlling endothelial cell proliferation and retinal neovascularization and therapies targeting neurofibromin-deficient EC may be beneficial.

AB - PURPOSE. Neurofibromatosis type 1 (NF1) is the result of inherited mutations in the NF1 tumor suppressor gene, which encodes the protein neurofibromin. Eye manifestations are common in NF1 with recent reports describing a vascular dysplasia in the retina and choroid. Common features of NF1 retinopathy include tortuous and dilated feeder vessels that terminate in capillary tufts, increased endothelial permeability, and neovascularization. Given the retinal vascular phenotype observed in persons with NF1, we hypothesize that preserving neurofibromin may be a novel strategy to control pathologic retinal neovascularization. METHODS. Nf1 expression in human endothelial cells (EC) was reduced using small hairpin (sh) RNA and EC proliferation, migration, and capacity to form vessel-like networks were assessed in response to VEGF and hypoxia. Wild-type (WT), Nf1 heterozygous (Nf1 +/- ), and Nf1 flox/+ ;Tie2cre pups were subjected to hyperoxia/hypoxia using the oxygen-induced retinopathy model. Retinas were analyzed quantitatively for extent of retinal vessel dropout, neovascularization, and capillary branching. RESULTS. Neurofibromin expression was suppressed in response to VEGF, which corresponded with activation of Mek-Erk and PI3-K-Akt signaling. Neurofibromin-deficient EC exhibited enhanced proliferation and network formation in response to VEGF and hypoxia via an Akt-dependent mechanism. In response to hyperoxia/hypoxia, Nf1 +/- retinas exhibited increased vessel dropout and neovascularization when compared with WT retinas. Neovascularization was similar between Nf1 +/- and Nf1 flox/+ ;Tie2cre retinas, but capillary drop out in Nf1 flox/+ ;Tie2cre retinas was significantly reduced when compared with Nf1 +/- retinas. CONCLUSIONS. These data suggest that neurofibromin expression is essential for controlling endothelial cell proliferation and retinal neovascularization and therapies targeting neurofibromin-deficient EC may be beneficial.

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KW - Ras

KW - Retinopathy of prematurity

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