EVOLUTION OF ANTIMETASTATIC ENZYME

描述(申请人提供)：癌症是一种毁灭性的疾病，几乎没有治愈方法。大多数治疗方法依赖于破坏细胞DNA或扰乱肿瘤细胞的细胞周期。转移性黑色素瘤尤其具有破坏性，因为它对常规化疗和放射治疗相对耐受，往往很快就会致命。然而，与黑色素瘤和其他癌症的肿瘤发生和转移有关的一个新途径是由氨基酸酪氨酸激活的。值得注意的是，黑色素瘤细胞系和小鼠模型中酪氨酸的耗尽抑制了肿瘤的侵袭和转移。酪氨酸的消耗可以通过限制饮食或通过酪氨酸苯酚裂解酶(TPL)的分解代谢来完成。KM测定的TPL活性很低，需要大量的酶用于进一步的动物研究或最终的人类治疗。“DNA改组”是一种能够通过基于DNA同源性的过程进化增强蛋白质的方法。然而，这种方法不适用于TPL，因为它的结构同源物在DNA水平上不相似。IntegriGen正在开发一种非同源“洗牌”基因的方法。将这种方法应用于第三方物流，以增强其Km测量的功能。因此，该项目有两个应用：1)进化一种新的具有增强活性的TPL变异体，该变异体随后可以被评估为抗癌治疗；2)建立一种新的方法来对非同源基因进行“改组”，作为一种普遍适用于蛋白质工程的分子进化技术。 建议的商业应用：不可用

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