Clinical Pediatric Dermatology Open Access

Stimulation of keratinocyte wound healing responses and re-epithelialization by novel epoxy-tiglianes via protein kinase C activation

Joint Event on 5^th International Conference on Advances in Skin, Wound Care and Tissue Science & 14^th International Conference on Clinical Dermatology

October 15-16, 2018 Rome, Italy

Rachael L. Moses, Glen M. Boyle, Paul Reddell, Robert Steadman and Ryan Moseley

Cardiff University, UK QIMR Berghofer Medical Research Institute, Australia QBiotics Ltd., Australia Welsh Kidney Research Unit, UK

Scientific Tracks Abstracts: Clin Pediatr Dermatol

Abstract:

Novel epoxy-tiglianes, EBC-46 and EBC-211, are sourced from seeds of the Fountain’s Blushwood Tree, indigenous to Queensland. EBC-46 possess potent tumouricidal properties, through classical PKC activation, and is under development by our industrial partner, QBiotics Ltd., as a human and veterinary anti-cancer pharmaceutical. In clinical studies, EBC-46 also stimulated exceptional dermal healing, manifested as accelerated wound re-epithelialisation, closure and minimal scarring. This work describes epoxy-tigliane effects on keratinocyte wound healing responses and their underlying mechanisms of action. Immortalized human epidermal keratinocytes (HaCaTs) were treated with EBC-46 or EBC-211 (0-10μg/ml). Cell cycle progression/proliferation were assessed by FACS analysis and MTT assay. HaCaT migration was assessed using in vitro scratch wound assays. Global gene expression changes induced by epoxy-tiglianes were quantified by Microarray analysis, with differentially expressed genes confirmed by protein level analysis. As epoxy-tiglianes mediate responses via classical protein kinase (PKC) activation, mechanistic studies were performed with BIM-1 (pan-PKC), Gö6976 (classical-PKC) and LY317615 (PKC-βI/PKC-βII) inhibitors. Western blotting confirmed phospho-PKC activation following epoxy-tigliane treatment. Both epoxy-tiglianes induced significant HaCaT cell cycle progression and proliferation; and also promoted significant HaCaT scratch wound closure. Microarray analyses identified key genes differentially expressed in EBC-46/EBC-211-treated HaCaTs, which contribute to their stimulatory effects on keratinocyte proliferation and migration. Enhanced proliferative and migratory responses were significantly abrogated by BIM-1 and Gö6976, although LY317615 exhibited minimal inhibitory effects. PKC activation increased following epoxy-tigliane treatment. Such findings explain the enhanced re-epithelialization responses in epoxy-tigliane-treated skin; and provide justification for their translational development as novel therapeutics for impaired wound re-epithelialisation.

Biography :

Rachael Moses completed her PhD at Cardiff University in 2016 and is currently a Postdoctoral Research Associate at Cardiff University, UK. Her research focuses on elucidating the mechanisms underlying the novel epoxy-tigliane pharmaceuticals exceptional dermal wound healing responses. She was awarded a travel bursary to visit a medical research institute in Queensland, Australia, to undertake Microarray Analysis determining key genotypic changes following epoxy-tigliane treatment. She has filed patents with an industrial partner in this sector (QBiotics Ltd.); and has been awarded conference prizes relating to this area.

E-mail: MosesR@cardiff.ac.uk