GLA is associated with ESCC progression and chemotherapy response via DNA damage repair-related pathways.

BACKGROUNDEsophageal squamous cell carcinoma (ESCC) is a highly lethal malignancy frequently complicated by chemoresistance. α-Galactosidase A (GLA), a lysosomal hydrolase associated with poor prognosis in multiple cancers, has not been investigated in ESCC.

METHODSTwo independent ESCC datasets (GSE161533, GSE38129) were used to screen commonly upregulated genes, which were ranked by diagnostic performance and further validated in an integrated single-cell RNA-seq atlas and an independent dataset (GSE20347), identifying GLA. GLA expression was validated by immunohistochemistry, qRT-PCR, and Western blot. Its biological function was assessed by siRNA-mediated knockdown combined with CCK-8, EdU, colony formation, Transwell, and wound healing assays. KEGG, GO, and GSEA were performed to explore underlying mechanisms. The effect of GLA knockdown on chemosensitivity, and the combination of the GLA pharmacological chaperone Migalastat with gemcitabine or cisplatin, were evaluated by CCK-8 assays in two ESCC cell lines.

RESULTSGLA was significantly upregulated in ESCC, showed strong diagnostic performance across datasets, and was specifically enriched in malignant epithelial cells at the single-cell level. Multiple validation approaches confirmed GLA overexpression in ESCC cell lines and patient tissues. GLA knockdown markedly suppressed proliferation, colony formation, and migration. Enrichment analyses suggested that high GLA expression was associated with DNA damage repair, cell cycle, and DNA replication-related transcriptional programs, as well as with gemcitabine/cisplatin resistance-related genes. GLA knockdown increased the sensitivity of ESCC cells to gemcitabine and cisplatin, and Migalastat combined with either agent enhanced cytotoxicity in two ESCC cell lines, with combination index analysis indicating synergy.

CONCLUSIONGLA is a novel, upregulated gene in ESCC with diagnostic relevance and an oncogenic role, and is associated with DNA damage repair-related transcriptional programs and chemotherapy response. Combining Migalastat with chemotherapy may represent a candidate strategy that warrants further mechanistic and in vivo investigation.