The overall goal of this Program Project is to develop improved vectors for gene delivery in vivo and the application of these vectors for treatment of cardiovascular disease in experimental animals. There are four research projects supported by four core units and complemented by two pilot and feasibility projects. The objective of Project l is to develop and test gutless adenoviral vectors using reporter genes, and to test of the utility of these vectors in the in vivo delivery of the low density lipoprotein receptor (LDLR) and the very low density lipoprotein receptor (VLDLR) genes in the treatment of hypercholesterolemia in experimental animals. In close collaboration with Project 2, helper- dependent adenoviral vectors will be produced and strategies will be developed to ensure long-term expression by transient immunosuppression, use of vectors of different serotypes, and use of gutless adenoviral vectors totally devoid of viral genes. Experimental animals will be normal mice, LDLR -/- mice and heterozygous LDLR-deficient rhesus monkeys. Project 2 will collaborate with Projects 1, 3, and 4 to develop gutless vectors for experiments in vitro and in vivo using helper-dependent viruses that contain viron protein genes from different serotypes. It will also develop integrating adenoviral vectors using a eukaryotic transposable element. Project 3 will apply a drug dependent regulatable system to produce transgenic mice that express LDLR that can be turned on by an exogenous ligand. It will optimize the system for possible use in humans in the future and to incorporate into helper-dependent gutless adenoviral vectors developed in Projects 1 and 2. Project 4 will test systematically the utility of adenoviral gene transfer to diminish the extent of apoptosis after coronary artery ligation, for five complementary classes of protein with proven ability to inhibit apoptosis in vitro. These projects will be supported by an Administrative Core, a Primate Core (at the Southwest Foundation for Biomedical Research where the rhesus monkeys are housed) which conducts the primate experiments ii collaboration with Projects 1, 2 and 3, and Vector Production Core, which produces large amounts of high quality gutless vectors for in vivo use. Two pilot and feasibility projects have been selected from eleven submissions for inclusion in this application. They deal with (i) control of antigen presentation in gene therapy and (ii) apoptosis-based gene therapy of vascular smooth muscle.
