The role of NEK2 and AURKB on triple-negative breast cancer progression

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, accounting for 10-20% of all cases. TNBC's aggressiveness is mostly attributed to its high rates of chromosomal instability, proliferation, and metastatic potential. Molecular subtyping has revealed that overexpression of mitotic kinases NEK2 and AURKB is implicated in chromosomal instability and epithelial-to-mesenchymal transition (EMT). Recent studies have shown that the mitotic kinases NEK2 and TTK (a kinase regulated by AURKB) are involved in early metastatic signaling by promoting EMT, cell migration, and invasion through distinct transcriptional pathways. Despite their clinical relevance, no co-targeting therapeutic strategy exists to interrupt the distinct mitotic signaling pathways driven by these kinases. Thus, our study aims to investigate the functional convergence of NEK2 and AURKB signaling pathways as drivers of EMT and metastatic progression in TNBC. We hypothesize that NEK2 and AURKB cooperatively regulate mitotic and EMT-associated phosphorylation events that fuel early metastasis. Our approach leverages TNBC cell models representing ancestrally distinct origins (MDA-MB-231 and MDA-MB-157) and utilizes genetic knockdown to dissect how individual and dual suppression of NEK2 and AURKB alters the phosphorylation state and functional behavior of TNBC cells. Preliminary findings using Kaplan-Meier survival analysis confirm that high expression of NEK2 or AURKB is significantly associated with overall survival and relapse-free survival. STRING (Search Tool for the Retrieval of Interacting Genes/Proteins) protein-protein interaction analysis showed that both AURKB and NEK2 may converge on EMT-related processes, while STRING-based GO enrichment suggested that their inhibition may impair metastatic progression by disrupting EMT signaling in TNBC. Western blot analysis confirmed a decrease in EMT biomarkers Vimentin and Slug after individual and dual knockdown of NEK2 and AURKB. Moreover, invasion assays demonstrated a statistically significant reduction after individual and dual knockdown of both kinases. Overall, these results suggest that suppression of the mitotic kinases NEK2 and AURKB could disrupt EMT-associated metastatic potential in TNBC MDA-MB-231 cells. Ongoing experiments will further confirm and expand these findings. This project may advance the development of more effective precision-targeted approaches for TNBC, particularly for African American and Caribbean Hispanic/Latino women who are disproportionately affected.