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Severe forms of EBS are associated with cytoplasmic protein aggregates, with elevated kinase activation of ERK1 and ERK2 (ERK1\/2; also known as MAPK3 and MAPK1, respectively), suggesting intrinsic stress caused by misfolded keratin protein. Human keratinocyte EBS reporter cells stably expressing GFP-tagged EBS-mimetic mutant K14 were used to optimize a semi-automated system to quantify the effects of test compounds on keratin aggregates. Screening of a protein kinase inhibitor library identified several candidates that reduced aggregates and impacted on epidermal growth factor receptor (EGFR) signalling. EGF ligand exposure induced keratin aggregates in EBS reporter keratinocytes, which was reversible by EGFR inhibition. EBS keratinocytes treated with a known EGFR inhibitor, afatinib, were driven out of activation and towards quiescence with minimal cell death. Aggregate reduction was accompanied by denser keratin filament networks with enhanced intercellular cohesion and resilience, which when extrapolated to a whole tissue context would predict reduced epidermal fragility in EBS patients. This assay system provides a powerful tool for discovery and development of new pathway intervention therapeutic avenues for EBS.<\/jats:p>","DOI":"10.1242\/jcs.258409","type":"journal-article","created":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T12:37:04Z","timestamp":1634128624000},"source":"Crossref","is-referenced-by-count":10,"title":["A cell-based drug discovery assay identifies inhibition of cell stress responses as a new approach to treatment of epidermolysis bullosa simplex"],"prefix":"10.1242","volume":"134","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2428-2308","authenticated-orcid":false,"given":"Tong San","family":"Tan","sequence":"first","affiliation":[{"name":"Skin Research Institute of Singapore, A*STAR, Immunos Building, 8A Biomedical Grove, Singapore138648"},{"name":"Institute of Medical Biology, A*STAR, Immunos Building, 8A Biomedical Grove, Singapore138648"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3280-7365","authenticated-orcid":false,"given":"John E. 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