The long term priority of this aspect of the Laboratory of Cancer Genetics is the development and implementation of methods integrating molecular and cytogenetic technologies in the study of cancer. The impetus for this effort is largely derived from the inadequacy of older technologies to deal with the complex changes in the structure of the genome and the myriad alterations of gene expression which occur during oncogenesis. Improvements in technology for placing molecular probes onto the cytogenetic map combined with the ability to convert cytogenetically observable structures into discrete molecular reagents now provide an important pathway for information flow between the molecular and cytogenetic arenas. Chromosome microdissection has provided a key technology for generating the reagents which facilitate this effort. This project is divided into tow major areas. 1) High-resolution positional reagents in visualization mehtods. [This work encompasses microdissection and amplification technology as well as high-resolution fluorescence in situ hybridization (FISH)]. 2) Isolation of region-specific cDNAs by microdissection-mediated cDNA capture. [This specific aim encompasses a variety of techniques aimed at using various hybrid selection techniques to identify candidate cDNAs involved in human disease]. A new technology, the use of cDNA microarrays, is being developed to allow simultaneous evaluation of cellular mRNA levels for thousands of genes. This technology specifically enables sensitive comparisons of gene transcript levels between cells from various pathological stages. (Experiments to date with a model system in which tumorigenic potential is genetically suppressed demonstrate that the method allows detection of many changes in gene expression associated with an important biological event.)
