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Skin Lipid Abnormalities in Patients with a History of Eczema Herpeticum Indicate the Involvement of the Sphingosine‐1‐Phosphate Signaling System in HSV Viral Replication
Author(s) -
Berdyshev Evgeny,
Goleva Elena,
Bronova Irina,
Jung John,
Bronoff Anna S.,
Norquest Kathryn,
Richers Brittany,
Leung Donald YM
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.654.1
Subject(s) - sphingosine , atopic dermatitis , ceramide , sphingosine 1 phosphate , sphingolipid , sphingosine kinase 1 , sphingosine kinase , pathogenesis , viral load , immunology , chemistry , medicine , biochemistry , virus , apoptosis , receptor
Rationale Atopic dermatitis (AD) with eczema herpeticum (EH) can result in systemic infections and can be life‐threatening. The purpose of this study was to identify differences in non‐lesional skin between AD with a history of recurrent EH (ADEH+), AD without EH (ADEH−), and non‐atopic (NA) controls to gain insight into mechanisms involved in EH pathogenesis using a novel minimally invasive skin tape strip (STS) methodology and mass spectrometric approaches. Methods STS layers 5–6 from 13 NA, 13 ADEH−, and 15 ADEH+ adult subjects were collected from non‐lesional skin. AD subjects were balanced by gender, age and disease severity; STS were subjected to lipidomic and filaggrin breakdown products analyses by liquid chromatography tandem mass spectrometry (LC‐MS/MS). For the in vitro experiments, differentiated primary human keratinocytes were transfected with control or sphingosine‐1‐phosphate (S1P) lyase (SGPL), sphingosine kinase‐1 (SPHK1), or sphingosine kinase‐2 (SPHK2) siRNA for 48h and infected with HSV‐1 (0.1 MOI). The effect of enzyme silencing on sphingolipidome was evaluated via the LC‐MS/MS. Viral replication was assessed in 24h with qPCR and plaque assay. Results ADEH+, as compared to ADEH− and NA, subjects had a significantly increased proportion of short‐chain over long‐chain NS‐ceramides (sphingosine N‐acylated with non‐hydroxy fatty acids) (p=0.002), and increased levels of sphingomyelins (p=0.006). In contrast to ADEH−, ADEH+ subjects demonstrated increased level of free C18‐ and C17‐sphingosines (p=0.007 and 0.052, correspondingly) that suggests upregulated skin sphingosine‐1‐phosphate (S1P) metabolism and signaling. A comparison of the effect of silencing of major enzymes that directly regulate S1P levels (SPHK1, SPHK2, SGPL) in primary human keratinocytes in vitro revealed that only the silencing of SGPL resulted in increased viral replication after keratinocyte inoculation with HSV‐1 (Mean±SE, 0.55±0.09 vs. 0.25±0.05 for S1PL vs. control siRNA transfected keratinocytes, p<0.01, as detected by qPCR). Conclusions STS from ADEH+ subjects and in vitro studies indicate involvement of the S1P signaling system in EH and suggest novel potential therapeutic targets to treat EH. Support or Funding Information This study was funded by the Atopic Dermatitis Research Network supported by NIH/NIAID grant U19 AI117673. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .