Caroline Pennington, Rob Nuttall, CJ Shukla, Sarah Porter, Clara Sampieri, Kostas Lazaridis, Madeleine Handsley, Angela Stokes, Mark Pitchers
We have established a quantitative real-time PCR (qPCR) assay that allows for rapid, reliable, and specific analysis of the RNA levels of numerous proteinases, inhibitors, growth factor, receptor, and regulatory proteins, in human- and mouse-derived cells and tissues. By analyzing each of these genes concurrently, we hope to identify: 1) potential prognostic indicators of disease; and 2) co-expression profiles that can be used to form hypotheses about the relationships between regulatory molecules with downstream effector functions such as proteolysis. Currently we have validated primer-probe sets for over 100 human genes including the complete MMP, ADAMTS and TIMP gene families. We are engaged in using expression profiling to analyses the “Degradome” – the complete repertoire of proteases that cells and tissues deploy – in a variety of developmental, repair and pathological states, including wound healing, cancer, multiple sclerosis and osteoarthritis. See also links to the FP6 CANCERDEGRADOME project.