Background
Ceramides play an essential role in the formation of the stratum corneum, and include a long-chain sphingoid base in an amide linkage with one or more fatty acids (FAs). Skin contains many unique FAs, particularly esterified omega (EO)-hydroxy(OH)ceramides with ultra-long FA chains, which are critical to the CLE.
Atopic Dermatitis, a common pediatric skin disorder with cutaneous inflammation and barrier impairment, is recognized to have a deficiency of ceramides, including EO-OH ceramides, based on targeted lipidomic analyses. Untargeted lipidomics (Q Exactive HF OrbiTrap) is a new technology that, as RNA-Seq or proteomics, allows comprehensive comparison of diseased vs healthy tissues but has yet to be described for skin.

Methods
We extracted lipids from tape strips of lesional atopic dermatitis (AD-L) and nonlesional (NL) skin from 31 AD patients (0.2-17.3 yrs old, mean 7 yrs; 51% female; 58% White) and 14 age- and site-matched healthy controls (C) for untargeted lipidomic analysis.

Results
PCA plots showed distinct separation between AD-L and C skin, regardless of sex or age. 637 distinct ceramide species were detected in AD and 616 in C skin, with 291 decreased in AD-L vs. C and 242 reduced in AD-L vs NL skin. Total normalized signal response of ceramides decreased 84% in AD-L and 51% in NL skin vs C (p < 0.001). Most ceramide classes decreased by >80% in AD-L vs C skin (EO by 88-93%; ODS/ADS 91%; NP 90%; NH 87%; NSD 83%; OH/AH 83%; and OSD/ASD 82%).
We found a new ceramide class (m), overall decreased in AD-L by 81%. A few species were increased in EOSD, OH/AH, OSD/ASD, NDS, NH, NP, and NS classes. Ceramide changes in NL skin were all in AD-L. Decreased AD-L ceramides correlated with eczema severity indices: SCORAD and EASI scores (Spearman r= –0.4 to -0.57, p < 0.001); m-ceramides with EASI and Pruritus ADQ scores (r= -0.43 and r= -0.44, p < 0.01). Increases of NDS in AD-L correlated with EASI (r=0.48, p < 0.01); and NH, NP, and NS with Itch VAS (r= 0.73 to 0.93, all p < 0.05).

Conclusion
The discovery of a new ceramide class (m), which is found to be decreased in AD-L highlights the value of untargeted lipidomic analyses for understanding skin disorders and in clinical trials that can target specific ceramides in the metabolic pathway.

Untargeted Lipidomic Analysis from Atopic Dermatitis Stratum Corneum Reveals Novel Ceramide Defects

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Untargeted Lipidomic Analysis from Atopic Dermatitis Stratum Corneum Reveals Novel Ceramide Defects

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