A preclinical model for skin sensitization prediction of antineoplasic drugs.

Abstract

e15643 Background: Skin side effects are common manifestations of antineoplastic drugs that are frequently observed in early clinical trials. Therefore, there is a need to identify skin toxic agents before clinical development in order to predict severe skin manifestations. In many cases, skin toxicity is due to sensitization, a key immunologic process mediating redness, swelling and itching that can lead to more severe skin alterations. Methods: We adapted three skin cellular in vitro techniques for cutaneous drug sensitization assessment of the Organization for Economic Cooperation and Development (OECD, 2012) in order to predict antineoplastic drug skin sensitization: 1) Direct peptide reactivity assay (DPRA) was assessed by high performance liquid chromatography (HPLC) coupled to ultraviolet detector, to detect covalent binding of antineoplastic to skin proteins (haptenation). 2) KeratinoSense skin sensitization assay was evaluated in keratinocyte cell line (HaCaT) transfected with a selectable plasmid to quantify luciferase gene induction by chemiluminescence as a measure of activation of Keap1-Nrf2-antioxidant response element involved in the cellular processes in skin sensitization and 3) The human cell line activation test (h-CLAT) represents the activation of dendritic cells by the increase of CD86 and CD54 surface markers measured by flow cytometry. An antineoplastic was considered sensitizing when at least two of these three tests were positive. Antineoplastic tested were paclitaxel, erlotinib, imatinib, sunitinib, cetuximab, olaparib, palbociclib and everolimus. Results: Paclitaxel was positive for all the three tests; the DPRA formation, keratinoSense activation and h-CLAT expression showing the highest level of sensitization. Moreover sunitinib, and imatinib were positive for two of the three in vitro cellular tests, being considered sensitizing drugs. In contrast, erlotinib, cetuximab, olaparib, palbociclib and everolimus were considered non-sensitizing antineoplastic agents. In the case of erlotinib a high proportion of apoptosis of the keratinocytes during the keratinosense test was observed, suggesting that cutaneous toxicity could be due to cytolysis instead of sensitization. Conclusions: Results obtained in this work suggest that pre-clinical assays may predict skin sensitization and be of potential value to predict skin sensitization of antineoplastic drugs before clinical development. Further assessment with in vivo tests will help to identify the cutaneous toxic mechanisms of those non-sensitizing drugs as cetuximab