Data Sheet 3_Identification and histological validation of autophagy-related core genes ADRB2 and PLK2 in keloids, with integrated immune infiltration analysis.pdf

IntroductionKeloids are pathological fibroproliferative scars characterized by excessive collagen deposition and a lack of effective targeted therapies. Autophagy dysregulation has been linked to keloid pathogenesis, but the underlying molecular mechanisms remain unclear.
MethodsTranscriptomic datasets were integrated and analyzed using differential expression analysis and weighted gene co-expression network analysis. Three machine learning algorithms—least absolute shrinkage and selection operator (LASSO), support vector machine–recursive feature elimination (SVM-RFE), and random forest—were applied to identify autophagy-related hub gene candidates in keloids. Immune infiltration and functional analyses were conducted to explore immune microenvironment alterations. Histological staining (H&E and Masson), immunohistochemistry, and Western blotting were used for tissue-level validation, while cellular experiments were performed in keloid fibroblasts with autophagy modulation.
ResultsADRB2 and PLK2 were consistently identified as key autophagy-related candidate genes. Immune-related analyses suggested that these genes may be involved in remodeling the keloid immune microenvironment by influencing the abundance and functional status of multiple immune cell subsets. Histological and protein-level assays demonstrated significantly higher expression of ADRB2 and PLK2 in keloid tissues compared with adjacent normal skin. In keloid fibroblasts, fibrotic markers (COL1/COL3) and autophagy-related markers (LC3-II/LC3-I and p62) were upregulated concomitantly with ADRB2 and PLK2 at baseline. Autophagy modulation altered ADRB2 expression (decreased with EBSS and increased with chloroquine), whereas PLK2 expression remained largely unchanged.
DiscussionThese findings identify ADRB2 and PLK2 as under-recognized autophagy- andimmunity-related candidate biomarkers in keloids, highlighting their potential relevance asdiagnostic indicators and future therapeutic research targets.