Reducing off target viral delivery in ovarian cancer gene therapy using a protease-activated AAV2 vector platform.

Gene therapy is a promising strategy for treating metastatic epithelial ovarian cancer (EOC). However, efficient vector targeting to tumors is difficult and off-target effects can be severely detrimental. Most vector targeting approaches rely on surface receptors overexpressed on some subpopulation of cancer cells. Unfortunately, there is no universally expressed cell surface biomarker for tumor cells. As an alternative, we developed an adeno-associated virus (AAV) based "Provector" whose cellular transduction can be activated by extracellular proteases, such as matrix metalloproteinases (MMP) that are overexpressed in the tumor microenvironments of the most aggressive forms of EOC. In a non-tumor bearing mouse model, the Provector demonstrates efficient de-targeting of healthy tissues, especially the liver, where viral delivery is <1% of AAV2. In an orthotopic HeyA8 tumor model of EOC, the Provector maintains decreased off-target delivery in the liver and other tissues but with no loss in tumor delivery. Notably, approximately 10% of the injected Provector is still detected in the blood at 24 h while >99% of injected AAV2 has been cleared from the blood by 1 h. Furthermore, mouse serum raised against the Provector is 16-fold less able to neutralize Provector transduction compared to AAV2 serum neutralizing AAV2 transduction (1:200 vs 1:3200 serum dilution, respectively). Thus, the Provector appears to generate less neutralizing antibodies than AAV2. Importantly, serum against AAV2 does not neutralize the Provector as well as AAV2, suggesting that pre-existing antibodies against AAV2 would not negate the clinical application of Provectors. Taken together, we present an EOC gene delivery vector platform based on AAV with decreased off-target delivery without loss of on-target specificity, and greater immunological stealth over the traditional AAV2 gene delivery vector.

Effect of age and cholestyramine feeding on rat liver 3-hydroxy-3-methyl glutaryl CoA reductase, sterol carrier protein 1 and sterol carrier protein 2 activities.

The rate of formation of sterol from squalene in livers from suckling rats was less than one-third that of adults. This difference was due to a lesser activity of microsomal enzymes in the suckling rat livers, and not to any difference in cytosolic sterol carrier protein 1. The microsomal enzymes and sterol carrier protein 2 of the cytosol required for the conversion 7-dehydrocholesterol to cholesterol were both lower in suckling rats compared to adults. Both those activities paralleled the differences in HMG-CoA reductase activities between suckling and adult rats. Feeding of cholestryamine to adult rats increased the activities of the microsomal enzymes, sterol carrier protein 1 and sterol carrier protein 2 involved in the conversion of squalene to cholesterol.

Financial prediction and trading strategies using neurofuzzy approaches.

Neurofuzzy approaches for predicting financial time series are investigated and shown to perform well in the context of various trading strategies involving stocks and options. The horizon of prediction is typically a few days and trading strategies are examined using historical data. Two methodologies are presented wherein neural predictors are used to anticipate the general behavior of financial indexes (moving up, down, or staying constant) in the context of stocks and options trading. The methodologies are tested with actual financial data and show considerable promise as a decision making and planning tool.