Validatie van moleculaire markers voor huidsensitisatie door "gene silencing" in gereconstrueerde huidmodellen
07 / 2009 - 07 / 2013
Keratinocytes are the first cells to be encountered by chemicals after skin contact and therefore considered a suitable target for studying the cutaneous response to sensitizers. In this project genome wide gene and protein expression analyses are employed as tools to discriminate between classes of chemicals using human cell lines and reconstructed skin models. This latter approach allows analysis of skin sensitization in an experimental model that recapitulates most of the in vivo characteristics and retains the actual human organotypic environment, while it shares the advantages of unrestricted experimental manipulation with cell-based assays. These analyses will deliver information on expression of m(i)RNAs and proteins that may discriminate between skin sensitization and skin irritation. In subsequent experiments, functional involvement of differentially expressed genes and proteins will be determined by specifically perturbing expression of m(i)RNAs of interest in skin models and cell lines and monitor effects on responses to chemicals. Furthermore, the markers identified by the experiments described above will be validated in a study involving human volunteers who will be exposed to known sensitizers (e.g. DCP) and from which skin biopsies will be analyzed for regulation of the markers identified by the studies described above using immunohistochemistry and quantitative PCR.