P38 Investigating the biosynthesis of 6-hydroxyceramides in the stratum corneum

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ID: 318471
2026
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Abstract
Abstract Introduction and aims The stratum corneum (SC) is a key component of the epidermal permeability barrier. It is composed of terminally differentiated keratinocytes in lipid lamellae, containing cholesterol, ceramides and free fatty acids. Ceramides are esters of sphingoid bases; to date, > 1200 species belonging to 12 classes were reported in SC. The ceramides are derivatives of sphingosine (S), dihydrosphingosine (DS), phytosphingosine (P), 6-hydroxysphingosine(H) and 4,14-sphingadiene (SD). Although ceramide biosynthesis has been studied, enzymes producing 6-hydroxysphingosine ceramides (CER[H]) are unknown. As SC ceramides are vital for skin barrier function, we explored potential biochemical pathways leading to CER[H]. Methods Rat epidermal keratinocyte cultures were air-lifted to form SC. Three dimensional (3D) cultures were treated with CER[N(8)DS(18)]; its metabolism to CER[N(8)S(18)], CER[N(8)P(18)], CER[N(8)H(18)] and CER[N(8)SD(18)] was monitored by lipidomics using ultrahigh performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UPLC/ESI-MS/MS) and multiple reaction monitoring. Cultures were treated with fenretinide [inhibitor of dihydroceramide desaturases 1/2]. Immunohistochemistry (ki67; keratin14) were used to assess the SC. Results 3D cultures formed SC as assessed by ki67 and keratin14 expression. UPLC/ESI-MS/MS ceramide profiling confirmed that 3D cultures formed all ceramides of interest, including CER[H]. Although CER[N(8)DS(18)] was metabolized to CER[N(8)S(18)] and CER[N(8)P(18)], CER[N(8)H(18)] was not detected. Fenretinide reduced utilization of CER[N(8)DS(18)] and impacted CER[N(8)S(18)] production, but did not induce CER[N(8)H(18)] formation. Conclusions Findings indicate that CER[H] species are not formed via metabolism of precursor CER[NDS]. Desaturation of CER[S] and CER[P] did not appear to be involved in CER[H] biosynthesis either. Alternative targets should be considered.
Reference Key
openalex_W7165682540 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Hawra Alsalman, Alexandra Kendall, Catherine O’Neill, Anna Nicolaou
Journal the british journal of dermatology
Year 2026
DOI
10.1093/bjd/ljag151.077
URL
https://doi.org/10.1093/bjd/ljag151.077
Keywords Keywords not found

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