Using a quantitative proteomics approach based on 4D-DIA combined with LC-MS/MS analysis, we identified TRPV2 as a potential host resistance factor that was significantly upregulated in DF-1 cells 48 hours post-DTMUV infection. Further experiments involving siRNA-mediated knockdown of TRPV2 expression in DF-1 cells revealed that suppression of TRPV2 expression inhibited DTMUV replication within the cells. Therefore, TRPV2 emerges as a promising candidate for poultry breeding programs aimed at enhancing resistance against DTMUV infection, serving as a potential target for the development of DTMUV-resistant animals in avian breeding.