Retinal Neuroprotection against Ischemic Injury Mediated by Intraocular Gene Transfer of Pigment Epithelium-Derived Factor

• Published in: Investigative ophthalmology & visual science Vol. 44; no. 10; pp. 4497 - 4504
• Main Authors: Takita, Hiroyasu, Yoneya, Shin, Gehlbach, Peter L, Duh, Elia J, Wei, Lisa L, Mori, Keisuke
• Format: Journal Article
• Language: English
• Published: Rockville, MD ARVO 01-10-2003; Association for Research in Vision and Ophtalmology
• Subjects: Adenoviridae - genetics; Animals; Apoptosis; Biological and medical sciences; Cell Count; Cell Survival; Cytoprotection - genetics; Enzyme-Linked Immunosorbent Assay; Eye Proteins; Female; Gene Transfer Techniques; Genetic Therapy; Genetic Vectors; In Situ Nick-End Labeling; Medical sciences; Nerve Growth Factors - genetics; Nerve Growth Factors - metabolism; Ophthalmology; Proteins - genetics; Proteins - metabolism; Rats; Rats, Inbred Lew; Reperfusion Injury - metabolism; Reperfusion Injury - pathology; Reperfusion Injury - prevention & control; Retinal Diseases - metabolism; Retinal Diseases - pathology; Retinal Diseases - prevention & control; Retinal Ganglion Cells - metabolism; Retinal Ganglion Cells - pathology; Retinopathies; Serpins - genetics; Serpins - metabolism; Retinopathy; Intraocular; Rat; Rodentia; Retina; Genetic transfer; Eye disease; Vertebrata; Mammalia; Reperfusion; Ischemia; Animal; Pigment epithelium-derived factor; Gene therapy
• ISSN: 0146-0404; 1552-5783; 1552-5783
• DOI: 10.1167/iovs.03-0052

To determine whether intraocular gene transfer of pigment epithelium-derived factor (PEDF) protects the retina from ischemia-reperfusion injury.
Four days before induction of pressure-induced ischemia, Lewis rats received intravitreous injection of 3 x 10(9) particles of an adenovirus vector expressing PEDF (AdPEDF.11) in one eye and 3 x 10(9) particles of an empty adenovirus vector (AdNull.11) in the contralateral eye. Seven days after reperfusion, eyes were enucleated and processed for morphometric analysis. Apoptotic cells stained by TdT-dUTP terminal nick-end labeling (TUNEL) in the retina were counted 12 hours after initiation of reperfusion. Retina levels of PEDF were measured by enzyme-linked immunosorbent assay.
PEDF levels in retinal homogenates from eyes receiving AdPEDF.11 injection were well above the background levels in the untreated baseline and control eyes (P = 0.04). Retinal thickness was preserved in AdPEDF.11-treated eyes. Retinal cell density was significantly greater in the ganglion cell layer (GCL; P = 0.014), inner nuclear layer (INL; P = 0.008), and outer nuclear layer (ONL; P = 0.008) of AdPEDF.11-treated eyes compared with the corresponding layers in AdNull.11-treated eyes. AdNull.11-treated eyes also had significantly more TUNEL-positive cells in these layers than AdPEDF.11-treated eyes (P < 0.05).
Adenoviral vector-mediated intraocular expression of PEDF significantly increases cell survival after ischemia-reperfusion injury of the retina. The protective effect may result from inhibition of ischemia-induced apoptosis. This study provides proof of concept for a gene transfer approach directed at interrupting programmed cell death induced by retinal ischemic insult.