Adenovirus (Ad) mediated gene transfer is a promising technology for the treatment of many genetic disorders of the Central Nervous System (CNS). Many of the drawbacks that exist with first-generation Ad vectors have been resolved through the use of 'gutted' or 'helper-dependent' vectors. The genetic basis of neurodegeneration is currently best understood in the retina. In addition, relative to the rest of the CNS, the physical accessibility of the eye makes the retina an excellent model system for studying gene therapy for the CNS. Upon ocular delivery, Ad vectors primarily infect the retinal pigment epithelium (RPE) and the Mueller cells of the retina. However, the diseases that most frequently cause blindness are associated with the expression of mutant proteins in the photoreceptor neurons. In our preliminary studies we have found that adenovirus-delivered green fluorescent protein fused to the full length Herpes Simplex Virus (HSV) tegument protein VP22 can translocate from infected cells to uninfected cells in culture or from the RPE to photoreceptors in vivo. This has led to the hypothesis we wish to test in this study: Can HSV VP22 be used to deliver therapeutic proteins to photoreceptor neurons via the RPE? This will be answered using a mouse model of inherited retinal degeneration (rd). Specifically, in this study we propose to 1) Construct an adenovirus vector expressing the protein transduction domains (PTD) of HSV VP22 fused to GFP and compare their ability to traffic GFP in cell culture and in murine retina. 2) Construct an adenovirus vector expressing a fusion between beta PDE and the PTD of HSV VP22. Assess the ability of this virus to express a functional PDE. 3) Administer a gutted adenovirus vector expressing either a VP22-beta PDE fusion to the retina of rd mice (which have a naturally occurring mutation in beta PDE) and assess the effects upon photoreceptor degeneration.

National Institute of Health (NIH)
National Eye Institute (NEI)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-GTIE (90))
Program Officer
Dudley, Peter A
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Utah
Schools of Medicine
Salt Lake City
United States
Zip Code
Gandhi, Jarel; Cashman, Siobhan M; Kumar-Singh, Rajendra (2011) Soluble CD59 expressed from an adenovirus in vivo is a potent inhibitor of complement deposition on murine liver vascular endothelium. PLoS One 6:e21621
Cashman, Siobhan M; Ramo, Kasmir; Kumar-Singh, Rajendra (2011) A non membrane-targeted human soluble CD59 attenuates choroidal neovascularization in a model of age related macular degeneration. PLoS One 6:e19078
Ma, Kelly N; Cashman, Siobhan M; Sweigard, J Harry et al. (2010) Decay accelerating factor (CD55)-mediated attenuation of complement: therapeutic implications for age-related macular degeneration. Invest Ophthalmol Vis Sci 51:6776-83
Sweigard, J Harry; Cashman, Siobhan M; Kumar-Singh, Rajendra (2010) Adenovirus vectors targeting distinct cell types in the retina. Invest Ophthalmol Vis Sci 51:2219-28
Read, Sarah Parker; Cashman, Siobhan M; Kumar-Singh, Rajendra (2010) A poly(ethylene) glycolylated peptide for ocular delivery compacts DNA into nanoparticles for gene delivery to post-mitotic tissues in vivo. J Gene Med 12:86-96
Read, Sarah P; Cashman, Siobhan M; Kumar-Singh, Rajendra (2010) POD nanoparticles expressing GDNF provide structural and functional rescue of light-induced retinal degeneration in an adult mouse. Mol Ther 18:1917-26
Johnson, Leslie N; Cashman, Siobhan M; Read, Sarah Parker et al. (2010) Cell penetrating peptide POD mediates delivery of recombinant proteins to retina, cornea and skin. Vision Res 50:686-97
Greenwald, David L; Cashman, Siobhan M; Kumar-Singh, Rajendra (2010) Engineered zinc finger nuclease-mediated homologous recombination of the human rhodopsin gene. Invest Ophthalmol Vis Sci 51:6374-80
Kumar-Singh, Rajendra (2008) Barriers for retinal gene therapy: separating fact from fiction. Vision Res 48:1671-80
Ramo, Kasmir; Cashman, Siobhan M; Kumar-Singh, Rajendra (2008) Evaluation of adenovirus-delivered human CD59 as a potential therapy for AMD in a model of human membrane attack complex formation on murine RPE. Invest Ophthalmol Vis Sci 49:4126-36

Showing the most recent 10 out of 14 publications