1079 Preliminary studies of targeted protein substitution in Netherton syndrome

Netherton syndrome is a severe subtype of congenital ichthyosis caused by mutations in SPINK5 encoding the lymphoepithelial kazal-type related inhibitor (LEKTI), serine protease that regulates activity kallikreins skin. Loss LEKTI leads to increased activity, resulting disrupted skin barrier and enhanced susceptibility infections. Current therapeutic options are mainly inadequate. Previously, recombinantly expressed domain 8+9 showed inhibitory properties on proteases vitro. Therefore, we aimed further investigate peptide for both stability assess its potential development topical protein replacement therapy. For evaluation purposes, cell culture models were established. Recombinant expression was performed E. coli. The purified freeze-dried. Characterization took place SDS-PAGE, immunoblotting size exclusion chromatography. lyophilized determined trypsin inhibition assay. Human epidermal equivalents(HEEs) cultured from primary keratinocytes healthy donors patients. H&E- immunostaining proteins including filaggrin adequate proliferation differentiation. assessed toluidine blue penetration We successfully produced 8+9, demonstrated structural stability, which could be confirmed at least 6 weeks 4 °C after reconstitution. HEEs suitable future efficiency testing substitution comparison with equivalents. Our study so far shows promising results utilization 8+9. With regard delivery formulations, suggest experiments encapsulation solid lipid nanoparticles or liposomes.