The thymidine phosphorylase (TYMP) gene encodes a pivotal enzyme involved in nucleoside metabolism, playing a critical role in the processing of chemotherapeutic agents. Despite its recognized importance, the influence of TYMP on tumor immunology and clinical outcomes remains largely unexplored. This study sought to elucidate the roles and regulatory pathways of TYMP across a spectrum of cancers and whether TYMP is related to DNA damage repair, thereby promoting stem cell maintenance, chemotherapy resistance and immunosuppression. Utilizing pan-cancer bulk sequencing and digital platforms, the study investigated the impact of TYMP on patient outcomes, genomic instability, cancer stemness, DNA repair, and immune cell dynamics. Moreover, single-cell datasets of TISCH, spatial transcriptomic data of SpatialDB, and multiple fluorescence staining were used to validate the association between TYMP expression and macrophages. In vitro viability (trypan blue), wound healing, Transwell, Cell cycle and M1 macrophage infiltration assays were performed to determine the biological functions of TYMP in Kidney renal clear cell carcinoma (KIRC) cells. Tools such as ROCplotter and cMap were employed to evaluate treatment responses and identify compounds targeting TYMP, while PDB and Autodock Vina facilitated structural modeling and binding simulations. TYMP was an oncogene in many cancer types. High TYMP was associated with lower genome stability and high expression of mismatch repair genes, stemness, homologous repair gene signature. Also, immune checkpoint CD276 was positively relevant to TYMP. Subsequently, we validated TYMP as the macrophage marker and showed its connection with other immunosuppressive cells and CD8+ T-cell depression. In vitro experiments showed that TYMP promoted migration, and invasion of KIRC cells and recruited the M2 macrophage to KIRC cells. Finally, potential TYMP -targeted drugs were screened out and docked to TYMP protein. TYMP was a novel oncogene, and it correlated with immunosuppression and DNA repair. TYMP was highly associated with immune checkpoint CD276 and was an macrophage biomarker in many cancers. This study will reveal TYMP's roles in pan-cancer and its potential as a novel therapeutic target, especially KIRC.Copyright © 2025 Elsevier B.V. All rights reserved.