%0 PDF %T Non-Viral Gene Therapy for the Treatment of Retinal Degeneration. %A Read, Sarah. %D 2017-04-14T13:40:54.657Z %8 2017-04-14 %R http://localhost/files/5h73q698k %X Abstract: Retinitis pigmentosa (RP) is the most common neurodegenerative disorder that leads to blindness and affects approximately 1 in 4000 individuals world wide. There is currently no curative therapeutic option for patients suffering from RP. Because of the chronic nature of RP and many other ocular diseases, gene therapy may offer an ideal form of treatment. We have shown that a novel synthetic peptide POD, or "protein for ocular delivery," is able to deliver small molecules to the retina in vivo. Generation of a POD-GFP fusion protein demonstrated that POD was able to deliver macromolecules to the retinal pigment epithelium (RPE) and photoreceptors after subretinal delivery, and to the retinal ganglion calls after intravitreal delivery. POD was also shown to efficiently compact DNA and deliver it to cells in vitro. This observation prompted us to develop use of POD as a non-viral vector in vivo. When conjugated with polyethylene glycol (PEG-POD), PEG-POD can compact plasmid DNA into 136 nm that can penetrate and transduce the retinal pigment epithelium (RPE) of adult murine retina in vivo. PEG-POD significantly increased expression of plasmid DNA 215-fold relative to DNA alone. PEG-POD can protect plasmid DNA from DNaseI digestion, resulting in significant transfection of the lung following intravenous injection. To observe the therapeutic implications of PEG-POD mediated transduction, nanoparticles containing an expression cassette for Glial Cell Line-Derived Neurotrophic Factor (PEG-POD~GDNF) were investigated for their ability to slow or inhibit blue light-induced photoreceptor apoptosis. Animals injected with PEG-POD~GDNF showed a significant decrease in apoptosis photoreceptor survival relative to control-injected animals. PEG-POD~GDNF injected eyes showed a 27-39% increase in their functional response relative to controls, as measured by electroretinogram. Taken together, PEG-POD was found to significantly increase gene delivery relative to both DNA alone and other PEGylated peptides. This is one of only two studies demonstrating both histological and functional rescue of a mouse model of retinal degeneration following non-viral administration of a therapeutic transgene into adult retina. Although rescue is short lived for clinical application, this study represents an important step in the development of non-viral gene therapy for retinal disease.; Thesis (Ph.D.)--Tufts University, 2012.; Submitted to the Dept. of Genetics.; Advisor: Rajendra Kumar-Singh.; Committee: Janis Lem, Gavin Schnitzler, Michael Rosenblatt, Naomi Rosenberg, and John Castellot.; Keywords: Genetics, and Ophthalmology. %[ 2022-10-11 %~ Tufts Digital Library %W Institution