Gene transfer of an engineered zinc finger protein enhances the anti-angiogenic defense system.
Zinc finger protein transcription factors (ZFP TFs) have been shown to positively or negatively regulate the expression of endogenous genes involved in a number of different disease processes. In this study we investigated whether gene transfer of an engineered ZFP TF designed to up-regulate expression of the chromosomal pigment epithelium-derived factor (Pedf) gene could suppress experimentally induced choroidal neovascularization (CNV). Transient transfection with engineered ZFP TFs significantly increased both Pedf messenger RNA (mRNA) and secreted PEDF protein levels in cell culture. Six weeks after intravitreous or subretinal injection of an adeno-associated viral (AAV) vector expressing the PEDF-activating ZFP TF in mice, we observed increased retinal Pedf mRNA, and a significant reduction in the size of CNV at Bruch's membrane rupture sites, assessed in vivo by fluorescein angiography or by postmortem measurements on choroidal flat mounts. Importantly, the anti-angiogenic activity persisted at 3 months after intravitreous injection. These data suggest that ZFP TF-driven enhancement of the endogenous anti-angiogenic defense system may provide a new approach for prophylaxis and treatment of neovascular diseases of the eye.
Yokoi
Katsutoshi
K
Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-9277, USA.
Zhang
Huanda Steve
HS
Kachi
Shu
S
Balaggan
Kamaljit S
KS
Yu
Qi
Q
Guschin
Dmitry
D
Kunis
Mike
M
Surosky
Richard
R
Africa
Liza M
LM
Bainbridge
James W
JW
Spratt
Sharon Kaye
SK
Gregory
Philip D
PD
Ali
Robin R
RR
Campochiaro
Peter A
PA
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D018919:Neovascularization, Physiologic / Q000235:genetics; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D012160:Retina / Q000378:metabolism; D015843:Serpins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D016335:Zinc Fingers; D000099174:Pigment Epithelium-Derived Factor
D005136:Eye Proteins; D009414:Nerve Growth Factors; D012333:RNA, Messenger; D015843:Serpins; D014157:Transcription Factors; D000099174:Pigment Epithelium-Derived Factor