Infectious laryngotracheitis virus (ILTV), formally known as Gallid alphaherpesvirus 1, represents a prominent alphaherpesvirus that poses a significant threat to the global poultry industry. Current routine vaccination strategies fail to eliminate latent infection. Host–pathogen interaction networks have become a key focus in antiviral research. Our previous study demonstrated that activation of the MEK/ERK signaling pathway upon ILTV infection restricts host cellular metabolic activity to mount protective host antiviral responses, yet the underlying molecular mechanism remains unclear. The present work systematically dissects the contribution of MEK/ERK signaling to intrinsic host defense against ILTV. The results show that ILTV infection activates the MEK/ERK pathway, which in turn promotes the expression, activation, and nuclear translocation of the transcription factor Fos. As a core transcriptional regulator, Fos represses host metabolic gene expression, thereby restricting viral replication and proliferation. Integrated multi-omics analyses further demonstrate that the MEK/ERK-Fos-metabolic regulatory axis operates uniformly during infection with avian, human, and porcine alphaherpesviruses, suggesting a broadly conserved host antiviral mechanism. This consistent cross species signaling pattern points toward an evolutionarily conserved host antiviral strategy and provides potential molecular targets for the development of broad-spectrum antiviral strategies against alphaherpesviruses.

Newcastle disease virus (NDV) is a promising oncolytic virus, but tumor heterogeneity constrains its clinical translation. The key determinants driving variable NDV oncolytic potency amid tumor heterogeneity remain incompletely elucidated. The main scope of this study included analyzing 22 human cancer cell lines representing 11 tumor types and identifying five gene co-expression modules associated with NDV susceptibility through weighted gene co-expression network analysis. Integrative multi-omics analyses of NDV-infected HCT116 cells, the least-susceptible cell line in our panel, predicted metabolic processes, innate immune defenses, and multiple forms of cell death as candidate pathways modulating NDV oncolytic efficacy. These key bioinformatic findings were further validated by genome-wide CRISPR-Cas9 screening, which uncovered POLR2G and SREBF1 as central components of the pro-oncolytic network and TNFRSF1A, TNFAIP3, and CASP9 as core mediators of the anti-oncolytic network. Their functional roles were further verified via genetic manipulation and pharmacological perturbation of JAK-STAT, TLR4, and apoptotic signaling. In conclusion, our findings support a bidirectional model wherein host metabolic programs promote NDV replication and oncolysis, whereas antiviral-innate immune signaling restricts them, providing candidate targets for host-directed combination therapy amid tumor heterogeneity.