Smart, Injury-Triggered Therapy for Ocular Trauma
Technical Report,15 Sep 2015,14 Sep 2016
University of British Columbia Vancouver Canada
Pagination or Media Count:
Traumatic eye injury TEI is one of the leading causes of monocular blindness in military personnel and young males worldwide. This profound and frequently irreversible posttraumatic loss of vision has a poor prognosis due to retinal cell death, scar formation, and lack of functional regeneration. Proliferative vitreoretinopathy PVR, a form of intraocular fibrosis, is often the primary reason for the loss of vision after ocular trauma, and frequently occurs after blunt trauma and open globe injuries caused by penetration, rupture, perforation, and presence of intraocular foreign bodies as well as after retinal re-attachment surgery. We genetically engineered protease activity sensor PAS as chimeric transmembrane protein that can respond to increase in metalloproteinase activity by sheddingreleasing tagged-ectodomains in the vicinity of affected cells after traumatic eye injury and induction of PVR. We demonstrated that upon infection with AAV carrying our construct, HEK293 cells and neurons in culture expressed the engineered HA-tagged PAS proteins. Their HA-tagged ectodomains were detected in the extracellular medium within minutes following stimulation with ionomycin and glutamate respectively. We used a rabbit PVR model of ocular trauma in which autologous blood was injected into the vitreous cavity of one eye after a surgical incision through the pars plana. We have successfully developed engineered adeno-associated virus AAV with CAG and CMV promoters, to transduce primary cortical neurons and retinal cells. These vectors will be applied in vivo to deliver therapeutic genes after ocular trauma.
- Medicine and Medical Research
- Anatomy and Physiology
- Genetic Engineering and Molecular Biology