Final Report Summary - SKINTERMINOMICS (Matrix metalloproteinase degradomics at the epidermal-dermal interface)
In this project, we exploited this specific advantage and modified TAILS to not only identify new substrates but also to monitor their cleavage over time. This significantly enhanced confidence in protease substrate discovery and helped to categorize cleavages based on clusters of processing events with different efficiencies. Next, we applied this improved technology to identify new MMP10 substrates in cell culture supernatants from keratinocytes and fibroblasts. Thereby, we identified novel MMP10-dependent cleavages of extracellular matrix proteins like type I collagen and ADAMTS-like protein 1 (ADAMTSL1) as well as of growth factor receptors, such as platelet-derived growth factor receptor alpha (PDGFRalpha). The functional importance of these cleavages for cellular behavior in healing skin wounds and skin tumors are currently under further investigation.
The second objective of this project aimed to better understand proteolysis under conditions of epidermal hyperproliferation. For this purpose, we used a mouse model of cutaneous wound healing and employed TAILS to examine protease cleavage products during skin repair at important steps of the healing process. Thereby, we related highly abundant proteases to cleavages at distinct time points after wounding and are currently investigating the functions of specific protease-substrate relations.
In conclusion, this project used and further developed the newest technologies to generate novel hypotheses for the elucidation of protease function in hyperproliferative epithelia. This opened up several avenues for future research, which are currently explored and will help to further explore proteases as drug targets to address unmet medical needs.