Isolation and Functional Characterization of Prostate Tumor-Specific Hypoxia-Inducible Promoter/Enhancer Elements for Use in Gene Therapy.

摘要

The major objective of this research project was to identify and characterize promoter/enhancer elements that can be used to specifically target the expression of therapeutic genes to hypoxic regions within prostatic tumors in vivo. We have characterized sequences present within the human TNF-alpha promoter that are responsible for both constitutive activity and induction by hypoxia. We have produced a mutated version of this promoter that exhibits substantially reduced constitutive activity in epithelial cells while retaining responsiveness to various activating stimuli including hypoxia and ionizing radiation. While lacking prostate specificity, it is hoped nevertheless that this element may prove useful in various cancer gene therapy applications. In addition, we have cloned and characterized a large panel of novel sequences that exhibit high levels of constitutive promoter activity in prostate cell lines. Finally, we have identified a number of hypoxia inducible-promoter elements and are continuing to characterize these with respect to their prostate specificity.