Research Project
2108264
Normal cell death regulates tumor and normal tissue kinetics, and its suppression may be associated with prognosis and diagnosis of cancer. Apoptosis is the normal physiological process of selective cell elimination entailing preprogrammed responses to physiological inducers and to a wide spectrum of chemotherapeutic drugs. A molecular hallmark of apoptosis is that genomic DNA is fragmented and remains inside the nucleus. For staining 3'-OH ends of the DNA strand breaks in situ, terminal deoxynucleotidyl transferase (TdT) and an affinity-tagged nucleotide are applied directly to a microscope slide. The chain is extended, and then a detectable antibody conjugate is bound. The current Phase I proposal aims to significantly advance the technology for in situ measurement of apoptosis by: (1) developing faster and more sensitive methods of conferring color on DNA strand breaks in situ ; and (2) using computerized data acquisition, digital confocal microscopy, and color image analysis to very rapidly translate staining into a quantitative apoptotic labeling index that is directly proportional to tumor cell kinetic rate. In Phase II research, the potential clinical utility of applying this system to answer medical questions about cancer treatment, diagnosis and prognosis will be assessed.
