Development of New Approaches for Breast Cancer Therapy and Diagnosis Based on Angiogenesis

摘要

This project focuses on angiogenesis, the growth of new blood capillaries, as a potential target for breast cancer therapy and diagnosis. Our approach was based on our finding that specialized cytoskeletal structures that form at sites of cell-extracellular matrix adhesion, known as the focal adhesion complexes (FACs), contain many of the signal transducing molecules that mediate the capillary growth-stimulating effects of angiogenic mitogens and extracellular matrix. During the last year, we have developed, screened, recloned, and begun to characterize monoclonal antibodies that were generated using isolated FACs as an immunogen. Three different hybridoma fusions have been carried out. These resulted in production of 385 different hybridoma lines of which 26% exhibited positive ELISA and immunofluorescence staining for FAC or cytoskeletal-associated proteins. Eight of these lines have been subcloned at least two times and three of these are currently being used in expression cloning to identify and isolate their molecular antigens. Identification of FAC- associated molecules in capillary cells should facilitate development of new angiogenesis inhibitors as well as alternative methods for breast cancer diagnosis.