PGK1 moonlights as an NF-kB/p65 kinase to orchestrate M2-like macrophage-mediated immunosuppression in triple-negative breast cancer

Triple-negative breast cancer (TNBC) is the most aggressive molecular subtype with limited therapeutic options. Metabolic reprogramming is a hallmark of cancer, and emerging evidence indicates metabolic enzymes possess non-canonical moonlighting functions. We identified phosphoglycerate kinase 1 (PGK1) as a key link between glycolysis and immunosuppression in TNBC. To investigate the role of PGK1 in shaping the tumor immune microenvironment, we performed single-cell RNA sequencing (scRNA-seq) on TNBC cells (4T1) with PGK1 knockdown (shPgk1) and negative control (shNC), each with three biological replicates. Single-cell libraries were prepared on the TaiM4 scRNA platform and sequenced on the BGI DNBSEQ platform using paired-end sequencing. A total of 33,138 high-quality cells were retained after quality filtering. Our analysis revealed that PGK1 knockdown significantly altered the transcriptional landscape and immune signaling pathways in TNBC cells.