The long-term goals of this research are to understand the biology of ovarian cancer and in that understanding test novel methods for treatment. The objective of this application are to establish an adenoviral (Ad) system of in situ gene transfer using a novel syngeneic mouse model of ovarian cancer and to assess the effects of inhibition of cellular Src kinase activity of progression on the disease. This model utilizes a stable cell line of mouse ovarian surface epithelial cells (ID8) that has undergone spontaneous in vitro transformation.
The first Aim will prepare the ID8 cells for subsequent experiments by producing green fluorescent protein expressing stable transfectants. In addition, Ad constructs containing DNA for betaGAL, a Src dominant negative protein and csk, an endogenous inhibitor of Src, will be prepared. The specific DNA expressed by the Ad will be under the control of L-plastin promoter, a promoter that will function to target the virus to cancerous ovarian epithelial cells. The infection efficiency of the Ad for the ID8 cells will e assessed. Inhibition of Src activity by Ad infection will be confirmed. In addition, the effects of inhibition of Src on in vitro cell proliferation, expression of plasminogen activator (PA) and vascular endothelial cell growth factor (VEGF) will be determined.
Aim 2 will assess the effects of AD-mediated inhibition of Src activity on progression of ovarian cancer in vivo in the mouse model. Initial experiments will assess the efficiency of Ad infection in vivo. Mice will be injected with ID8 cells and after 8 weeks when macroscopic tumors are present animals will be treated with Ad expressing beta GAL. BetaGAL expression in tumors and normal tissues will be assessed. In a similar experiment, the effects of in vivo treatment with the specific Src-inhibitory Ad on tumor Src activity, expression of PA and VEGF will be determined. The final set of experiments in this Aim will assess the effects of inhibition of Src by in vivo Ad infection on overall progression of the disease. Treatment with the virus will be tested at several time points after injection of ID8 cells.
Aim 3 will assess the effects of combined Ad-mediated inhibition of tumor Src activity with taxol and cisplatin treatment on the progression of the cancer in the mouse. Ad and drug treatments will be given at various times during the progression of the cancer.
The final aim, 4, will focus on the discovery of new drugs and drug combinations specific for killing ovarian cancer cells with high and low levels of Src expression. Studies in this proposal will provide novel insight into the role of Src in ovarian cancer and test the potential of inhibition of Src activity as a potential therapy.
Showing the most recent 10 out of 240 publications
