Mechanisms of Met Induced Hepatocytes Survival

点击翻译按钮获取中文摘要

DESCRIPTION (provided by applicant): Escape from apoptosis is a cardinal feature of cancer cells derived from liver tumors and other malignancies. The Hepatocyte Growth Factor Receptor (HGFR) known as Met is a transmembrane receptor tyrosine kinase (RTK) that plays an important role in promoting hepatocyte survival and proliferation, and it is overexpressed in various human carcinoma tissues and cell lines including hepatocellular carcinoma (HCC). Understanding the molecular mechanisms of HCC development is a key NIDDK initiative (i.e., the Action Plan for Liver Disease Research). Using structure-function studies, we discovered that the C-terminal end of human Met's intracellular domain harbors a novel tandem Caspase-3 cleavage site which we named the Met Caspase Decoy Site (MCDS). It has the following sequence: DNAD(DEVD(TRPASFWETS ((denotes the caspase cleavage site). Our central hypothesis to be tested in this A2 Renewal Application is that, by virtue of harboring a unique caspase substrate decoy motif (MCDS), the cytoplasmic tail of human Met functions as a 'bait' and traps the active site of Caspase-3 by forming a stable transition state intermediate during the cleavage process. This results in inhibition of Caspase-3 activity, apoptosis resistance and promotion of HCC. State-of-the-art molecular biochemical and biological approaches are planned to test this intriguing hypothesis. In Aim 1, we will investigate the functional role of the Met Caspase Decoy Site (MCDS) in Caspase-3 inhibition and promotion of hepatocyte survival and hepatocarcinogenesis using loss-of-function and gain-of-function approaches. In Aim 2, we will assess the molecular mechanisms involved in MCDS-Caspase-3 interaction and Caspase-3 inhibition. In Aim 3, we will determine whether MCDS function depends on the activation status of Met. From these studies, we should gather substantial insight into a novel pro-survival mechanism never before described.