7-Ketocholesterol promotes T cell migration through Ca(2+)-NFATc1 pathway-mediated F-actin polymerization and proinflammatory cytokine production in oral lichen planus.

BACKGROUND: Oral lichen planus (OLP) is a chronic T-cell-mediated inflammatory disorder of unknown etiology. Accumulating evidence has demonstrated elevated cholesterol levels in OLP, and its oxidation products --oxysterols have been implicated in T cell dysfunction. However, whether the oxysterol is involved in OLP pathogenesis remains to be fully elucidated. METHODS: Metabolomics was performed to profile oxysterols in the plasma of OLP patients, followed by functional enrichment analysis. Single-cell RNA sequencing was utilized to characterize gene expression dysregulation in tissue-resident T cells isolated from OLP lesions. Flow cytometry, immunofluorescence, and qRT-PCR were collectively used to quantify Ca(2+) concentration, cell apoptosis, protein expression, intracellular signaling, and gene transcription levels. Functional validation was conducted through a co-culture model and Transwell migration assays to assess the cytotoxic and migratory capacity of OLP T cells. RESULTS: The oxysterol profiles were aberrant in OLP plasma, with marked accumulation of 7-ketocholesterol (7KC). Functional analysis identified significant enrichment of differential metabolites in androstenedione metabolism. 7KC upregulated the expression of cholesterol regulators (SREBP2/LXR) in OLP T cells. Pro-7-ketocholesterogenic gene sets were dysregulated in OLP tissues, with localized T cells exhibiting enriched Ca(2+) -NFATc1 signaling and coordinated F-actin polymerization/ITGAL (LFA-1α) upregulation, positively correlating with migration signatures. Peripheral OLP T cells showed elevated Ca(2+), nuclear NFATc1, F-actin polymerization, and LFA-1α, all of which, along with ITGAL/IL1B/CCL4/IL6 levels, were further potentiated by 7KC treatment. 7KC was confirmed to enhance migrations of primary OLP T cells and OLP plasma-pretreated Jurkat T cells toward LPS-treated keratinocytes, without affecting keratinocyte apoptosis. Furthermore, CM4620-mediated blockade of Ca(2+) -NFATc1 pathway in OLP T cells inhibited 7KC-induced NFATc1 activation, reduced the expressions of F-actin and its modulators ACTB/DIAPH1, and IL1B/CCL4/IL6 gene expressions, with migration suppressions of both primary OLP T cells and OLP plasma-pretreated Jurkat T cells. CONCLUSIONS: 7KC could promote T cell migration through Ca(2+) -NFATc1 pathway-mediated F-actin polymerization and expression of IL1B/CCL4/IL6 in OLP.