title: Short hairpin RNA-mediated knockdown of VEGFA in Müller cells reduces intravitreal neovascularization in a rat model of retinopathy of prematurity.
abstract: Vascular endothelial growth factor (VEGF) A is implicated in aberrant angiogenesis and intravitreous neovascularization (IVNV) in retinopathy of prematurity (ROP). However, VEGFA also regulates retinal vascular development and functions as a retinal neural survival factor. By using a relevant ROP model, the 50/10 oxygen-induced retinopathy (OIR) model, we previously found that broad inhibition of VEGFA bioactivity using a neutralizing antibody to rat VEGF significantly reduced IVNV area compared with control IgG but also significantly reduced body weight gain in the pups, suggesting an adverse effect. Therefore, we propose that knockdown of up-regulated VEGFA in cells that overexpress it under pathological conditions would reduce IVNV without affecting physiological retinal vascular development or overall pup growth. Herein, we determined first that the VEGFA mRNA signal was located within the inner nuclear layer corresponding to CRALBP-labeled Müller cells of pups in the 50/10 OIR model. We then developed a lentiviral-delivered miR-30eembedded shRNA against VEGFA that targeted Müller cells. Reduction of VEGFA by lentivector VEGFA-shRNAetargeting Müller cells efficiently reduced 50/10 OIR up-regulated VEGFA and IVNV in the model, without adversely affecting physiological retinal vascular development or pup weight gain. Knockdown of VEGFA in rat Müller cells by lentivector VEGFA-shRNA significantly reduced VEGFR2 phosphorylation in retinal vascular endothelial cells. Our results suggest that targeted knockdown of overexpressed VEGFA in Müller cells safely reduces IVNV in a relevant ROP model.
authors: The John A. Moran Eye Center, The University of Utah, Salt Lake City, Utah, USA.
authors: Haibo
authors: H
authors: Wang
authors: George W
authors: GW
authors: Smith
authors: Zhihong
authors: Z
authors: Yang
authors: Yanchao
authors: Y
authors: Jiang
authors: Manabu
authors: M
authors: McCloskey
authors: Kenneth
authors: K
authors: Greenberg
authors: Pete
authors: P
authors: Geisen
authors: William D
authors: WD
authors: Culp
authors: John
authors: J
authors: Flannery
authors: Tal
authors: T
authors: Kafri
authors: Scott
authors: S
authors: Hammond
authors: M Elizabeth
authors: ME
authors: Hartnett
mesh_terms: D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology; D063928:Ependymoglial Cells / Q000378:metabolism* / Q000473:pathology*; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D051381:Rats; D015861:Retinal Neovascularization / Q000235:genetics / Q000473:pathology*; D012178:Retinopathy of Prematurity / Q000378:metabolism / Q000473:pathology*; D015398:Signal Transduction; D013194:Staining and Labeling; D014161:Transduction, Genetic; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
chemical_list: C020932:11-cis-retinal-binding protein; D002352:Carrier Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat
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