Identifying the therapeutic effects of talniflumate in metastatic papillary thyroid microcarcinoma through integrating multiple omics analysis.

Papillary thyroid microcarcinoma (PTMC) is a prevalent endocrine malignancy, and metastatic progression remains a major clinical concern. Identifying novel therapeutic strategies and metastasis-associated targets is therefore urgently needed. Integrated transcriptomic analyses were performed using TCGA and GEO datasets to identify metastasis-associated genes and biological pathways in PTMC. Functional enrichment, immune infiltration analysis, single-cell RNA sequencing analysis, and external validation were conducted to characterize candidate genes. The connectivity map (CMap) database was used for drug screening, followed by network pharmacology, molecular docking, and molecular dynamics simulations to evaluate potential therapeutic candidates. In vitro assays, including proliferation, migration, invasion, macrophage co-culture experiments, and Western blot analysis, were performed to evaluate the biological effects of the candidate drug. Twenty differentially expressed genes were identified between metastatic and non-metastatic PTMC samples, among which AREG and HBEGF were prioritized as hub candidates. HBEGF showed a negative correlation with tumor mutational burden, although its biological significance requires further investigation. Enrichment analyses suggested that ErbB signaling may be involved in PTMC metastatic progression. External validation using an independent GEO cohort provided additional support for the association between HBEGF expression and metastatic characteristics. CMap-based screening identified talniflumate as a potential therapeutic candidate. In vitro experiments demonstrated that talniflumate suppressed proliferation, migration, and invasion of thyroid cancer cells. Furthermore, talniflumate promoted M1 macrophage polarization and increased pro-inflammatory cytokine secretion in a cancer cell-macrophage co-culture model. Western blot analysis showed reduced HBEGF protein expression following talniflumate treatment, suggesting a potential regulatory effect on the HBEGF-associated signaling network. This study identifies HBEGF as a potential metastasis-associated candidate in PTMC and suggests that talniflumate may represent a promising therapeutic candidate. The proposed HBEGF-associated regulatory mechanism provides a potential direction for further investigation, while larger independent cohorts and additional mechanistic studies are required for validation.