A METHOD TO IDENTIFY UPSTREAM REGULATORS OF ONCOGENES

DESCRIPTION (provided by applicant): A comprehensive understanding of all transduction partners, transcription factors, and their targets responsible for each type of primary tumor is lacking, despite intense effort. Furthermore, only a limited number of known "points of regulatory convergence," covering multiple cancer types, exists. Vaccinex has developed a highly efficient, proprietary technology for the identification of genes based on functional selection within a mammalian background. This method utilizes cDNA libraries inserted into the vaccinia virus (vv) genome and has been successfully applied to the isolation of novel tumor antigens. The ability to make libraries in vaccinia virus represents a major advance, since vaccinia virus is the only mammalian expression vector in which it is possible to construct a diverse and representative cDNA library whose elements can be efficiently recovered from the cytoplasm of infected cells. We propose to apply a variant of this functional gene selection technology to use known oncogenes and tumor cell markers as 'bait' to identify proteins that modulate their activity. This approach represents a refreshing, functional alternative to global transcript monitoring experiments performed with cDNA and oligonucleotide microarrays. Specifically, where microarray experiments monitor global transcriptional events related to cellular environment, our technique chooses a well-defined end point of transformation, either in the form of an up-regulated transcript or an over-abundant protein and proposes to uncover proteins that regulate these targets. Vaccinex technology, therefore, has the unique capability to functionally scan upstream in a transduction pathway. Moreover, the technique is designed to identify common points of regulation from overlapping pathways. Defining these regulatory mechanisms directly in the context of cancer is a critical step for generating more effective therapeutics.