EVOLUTION OF ANTIMETASTATIC ENZYME

DESCRIPTION (provided by applicant): Cancer is a devastating disease with few cures. Most treatments rely on damaging cellular DNA or disrupting the cell cycle in tumor cells. Metastatic melanoma is especially devastating, as it is often rapidly fatal due to its relative resistance to conventional chemo and radiation therapy. However, a novel pathway involved in tumorigenesis and metastasis in melanoma and other cancers is activated by the amino acid tyrosine. Remarkably, tumor invasion and metastases are suppressed by depletion of tyrosine in melanoma cell lines, as well as mouse models. Tyrosine depletion can be accomplished by dietary restriction or through catabolism by the enzyme tyrosine phenol-lyase (TPL). The activity of TPL as measured by Km is low, requiring large amounts of the enzyme for further animal investigation or eventual human therapy. "DNA shuffling" is one method capable of evolving enhanced proteins through a DNA homology based process. This method, however, is not applicable to TPL because its structural homologs are not similar at the DNA level. IntegriGen is developing a method to non-homologously "shuffle" genes. This method will be applied to TPL in order to enhance its function as measured by Km. Thus, this project has two applications: 1) to evolve a novel TPL variant which has enhanced activity, that can subsequently be evaluated as an anti-cancer therapeutic, and 2) to establish a new method to non-homologously "shuffle" genes as a molecular evolution technology applicable in general to protein engineering. PROPOSED COMMERCIAL APPLICATIONS: NOT AVAILABLE