The common coxsackie-adenovirus receptor protein (CAR) mediates adenoviral infection. Replication deficient recombinant adenoviruses (rAv) are potent vectors for DNA transfer (transduction) in gene therapy. We have developed a method for delivering rAv to the glomerulus for the purpose of treating diabetic glomerulosclerosis. Understanding the role of CAR in this process is essential for refining rAv-based approaches to glomerular gene therapy. Thus, the primary goal of this project is to assess the role of CAR in mediating adenoviral transduction in the normal and diabetic glomerulus. The work has 3 specific aims.
Aim #1 will assess CAR expression in vitro using Northern analysis and Western blotting. In addition, the function of CAR in mediating transduction will be assessed using specific anti-CAR antibody.
Aim #2 will define in vivo expression of CAR using immunohistochemistry, Northern analysis and Western blotting. Glomerular and cellular localization of CAR will be investigated using laser microdissection. CAR-mediated transduction in vivo will be assessed using anti-AR antibodies and transduction with rAv.
Aim #3 will test the effect of a diabetic milieu on expression and function of CAR in vitro and in vivo. Taken together, the results of these studies will help clarify glomerulosclerosis BAR expression and function. and allow refinement of adenovinal-based approaches to the gene therapy of diabetic glomeruloscerosis.
|Bhatt, Udayan Y; Sferra, Thomas J; Johnson, Amy et al. (2002) Glomerular beta-galactosidase expression following transduction with microsphere-adenoviral complexes. Kidney Int 61:S68-72|