Integrative transcriptomic and single-cell analyses link ENO1 to CD4+ T cell function and glycolysis-associated metabolism in systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by immune dysregulation and cellular metabolic abnormalities, including enhanced glycolytic activity in CD4 + T cells. However, the molecular regulators that link metabolic alterations to aberrant CD4 + T cell responses remain incompletely understood. We integrated bulk RNA sequencing (RNA-seq) datasets from peripheral blood mononuclear cells (PBMCs) of SLE patients with a set of glycolysis- and lactate metabolism-related genes (GALMRGs) to prioritize candidate metabolic regulators. Candidate genes were then evaluated in independent bulk RNA-seq, single-cell RNA sequencing (scRNA-seq), and purified CD4 + T cell datasets, among which ENO1 emerged as a leading hit. To investigate its role in CD4 + T cell function, we performed in silico perturbation analysis and validated predictions using clinical samples and in vitro experiments in human and murine CD4 + T cells. ENO1 was reproducibly upregulated across multiple independent cohorts and emerged as the top candidate. scRNA-seq revealed elevated GALMRG scores in CD4 + T cells from SLE patients. In silico perturbation linked ENO1 to T-cell activation programs, and experimental ENO1 knockdown altered the expression of activation-related genes, including CCR7 , IL7R , JUNB , and AIF1 . At the protein level, ENO1 was elevated in PBMCs and CD4 + T cells from SLE patients, and ENO1 expression in CD4 + T cells correlated positively with SLE Disease Activity Index (SLEDAI) scores. Functionally, ENO1 inhibition suppressed proliferation of both human and murine CD4 + T cells, and ENO1 knockdown reduced glucose uptake and intracellular lactate levels in activated murine CD4 + T cells. Molecular docking identified Barasertib and CC_401 as candidate ENO1-interacting compounds, providing a computational basis for future validation. This study identifies ENO1 as a glycolysis- and lactate metabolism-associated factor associated with CD4 + T cell function in SLE and offers a resource for further mechanistic investigation of glycolysis and lactate metabolism in the disease.