title: Gene therapy for traumatic central nervous system injury and stroke using an engineered zinc finger protein that upregulates VEGF-A.
abstract: Recent studies have identified anti-apoptotic functions for vascular endothelial growth factor (VEGF) in the central nervous system (CNS). However, VEGF therapy has been hampered by a tendency to promote vascular permeability, edema, and inflammation. Recently, engineered zinc finger proteins (ZFPs) that upregulate multiple forms of VEGF in their natural biological ratios, have been developed to overcome these negative side effects. We used retinal trauma and ischemia models, and a cortical pial strip ischemia model to determine if VEGF upregulating ZFPs are neuroprotective in the adult CNS. Optic nerve transection and ophthalmic artery ligation lead to the apoptotic degeneration of retinal ganglion cells (RGCs) and are, respectively, two highly reproducible models for CNS trauma or ischemia. Adeno-associated vectors (AAV) vectors encoding VEGF-ZFPs (AAV-VEGF-ZFP) significantly increased RGC survival by ∼twofold at 14 days after optic nerve transection or ophthalmic artery ligation. Furthermore, AAV-VEGF-ZFP enhanced recovery of the pupillary light reflex. RECA-1 immunostaining demonstrated no appreciable differences between retinas treated with AAV-VEGF-ZFP and controls, suggesting that AAV-VEGF-ZFP treatment did not affect retinal vasculature. Following pial strip of the forelimb motor cortex, brains treated with an adenovirus encoding VEGF ZFPs (AdV-ZFP) showed higher neuronal survival, accelerated wound contraction, and reduced lesion volume between 1 and 6 weeks after injury. Behavioral testing using the cylinder test for vertical exploration showed that AdV-VEGF-ZFP treatment enhanced contralateral forelimb function within the first 2 weeks after injury. Our results indicate that VEGF ZFP therapy is neuroprotective following traumatic injury or stroke in the adult mammalian CNS.
authors: Department of Surgery, University of Toronto, Toronto, Ontario, Canada.
authors: Philippe M
authors: PM
authors: D'Onofrio
authors: Mahinthan
authors: M
authors: Thayapararajah
authors: Meghan D
authors: MD
authors: Lysko
authors: Mark
authors: M
authors: Magharious
authors: S Kaye
authors: SK
authors: Spratt
authors: Gary
authors: G
authors: Lee
authors: Dale
authors: D
authors: Ando
authors: Richard
authors: R
authors: Surosky
authors: Michael G
authors: MG
authors: Fehlings
authors: Paulo D
authors: PD
authors: Koeberle
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000502:physiology; D001930:Brain Injuries / Q000235:genetics / Q000628:therapy*; D015316:Genetic Therapy / Q000379:methods*; D020221:Optic Nerve Injuries / Q000235:genetics / Q000628:therapy; D015202:Protein Engineering; D051381:Rats; D020127:Recovery of Function / Q000235:genetics; D020521:Stroke / Q000235:genetics / Q000628:therapy*; D016896:Treatment Outcome; D015854:Up-Regulation / Q000235:genetics*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D042461:Vascular Endothelial Growth Factor A