Glycolytic reprogramming in host response to Borrelia burgdorferi: A gene signature revealed by integrative bioinformatics analysis and machine learning.

Lyme disease (LD), a multifaceted condition caused by Borrelia burgdorferi (Bb), remains poorly understood, particularly regarding metabolic pathways. This study aimed to evaluate the role of glycolysis-related genes (GRGs) in LD pathogenesis and identify key genes and mechanisms relevant to diagnosis and therapy. Differentially expressed GRGs were identified and further analyzed by correlation analysis and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. Key genes were screened using least absolute shrinkage and selection operator (LASSO) and support vector machine-recursive feature elimination, followed by gene set enrichment analysis, gene set variation analysis and immune infiltration analysis using CIBERSORT. Diagnostic value was assessed by receiver operating characteristic and nomogram analyses, and a competing endogenous RNA network and protein-drug interaction predictions were constructed. The expression of key genes was further validated by reverse transcription-quantitative PCR (RT-qPCR) in Bb-infected THP-1 cells, and glucose and lactate concentrations in the culture supernatants were measured using commercial colorimetric assay kits. A total of 63 differentially expressed GRGs were identified. Of note, two key genes [lactate dehydrogenase A (LDHA) and thioredoxin (TXN)] exhibited a strong diagnostic performance. Immune infiltration analysis revealed that these genes were associated with regulatory T cells (r=0.33, P=0.05), gamma delta T cells (r=-0.34, P=0.042), CD4 memory resting T cells (r=-0.4, P=0.016) and monocytes (r=-0.36, P=0.03). The potential glycolysis-targeting drugs were predicted. RT-qPCR analysis revealed increased LDHA and TXN expression in Bb-infected THP-1 cells. In addition, Bb stimulation reduced extracellular glucose levels and increased lactate accumulation in culture supernatants. Overall, this integrative analysis revealed notable alterations in glycolytic genes during Bb infection, suggesting that dysregulation of glycolysis contributes to LD immunopathology. The identified key genes (LDHA and TXN) may serve as infection-related transcriptional response markers, offering insights into LD mechanisms and potential precision-based strategies.