Mapping the function of EF-hand domain-containing protein 2 and determining its clinical relevance in non-small-cell lung cancer through single-cell transcriptomics

Objective: This study aims to explore the cellular composition and transcriptional variability within the tumor microenvironment (TME) of non-small-cell lung cancer (NSCLC) using single-cell RNA sequencing (scRNAseq) and to assess the role of EF-hand domain-containing protein 2 (EFHD2) in tumor progression and its involvement in relevant signaling pathways. Material and Methods: We analyzed scRNA-seq datasets to map the cellular and transcriptional landscape of NSCLC tumors. Immunohistochemistry (IHC) was employed to validate the expression of EFHD2 and assess immune cell infiltration in clinical samples. To further investigate the functional effect of EFHD2, we performed Western blot and quantitative real-time polymerase chain reaction analyses as well as cell proliferation, migration, invasion, and apoptosis assays. We also explored the janus kinase (JAK)-signal transducers and activators of transcription (STAT) signaling pathway as a potential underlying mechanism. Results: The scRNA-seq analysis revealed that epithelial cells were the predominant population within the TME, alongside endothelial cells, fibroblasts, macrophages, and a small proportion of stem cells. EFHD2 expression exhibited considerable variability, with higher levels associated with clusters enriched in transcriptionally active and immunomodulatory pathways. The IHC results demonstrated elevated EFHD2 expression and immune cell infiltration in tumor tissues compared with adjacent non-tumor tissues. Conclusion: EFHD2 expression in the NSCLC TME correlates with immune cell infiltration and may play a significant role in tumor progression and immune modulation. The JAK-STAT signaling pathway may be a potential mechanism underlying the effect of EFHD2. This work provides a new avenue for targeted therapy in NSCLC.