In Vivo Estradiol, Tamoxifen and Raloxifene Modulation of Association/ Dissociation Kinetics for Estrogen Receptor, Interacting Co-Factors and DNA binding Sites

摘要

This one-year, concept award was approved for the development of quantitative fluorescence imaging technologies that measure ligand-regulated co- localization and interactions of estrogen receptor with its interacting factors. DAMD support enabled us to successfully quantify the extent of factor co- localization using computer-assisted techniques. We also applied these quantitative techniques to accurately measure the extent of energy transfer, between fluorophores as a marker of the extent of interaction between attached molecules. One manuscript was published (see Appendix I) that detailed the effects of estradiol, tamoxifen, and four other ligands on the direct interactions of estrogen receptor with specific protein targets in living cells. A second manuscript is currently being written that expands this analysis to more interacting targets and more ligands. To date, we have used these technologies to identify at least two novel estrogen receptor ligands that regulate estrogen receptor interactions distinct from those regulated by current Selective Estrogen Receptor Modulators, including tamoxifen and raloxifene. Thus, DAMD support permitted the successful development of technologies that dramatically improve the identification and characterization of next-generation, anti-breast cancer reagents.