Synthetic Lethality Screening Highlights Colorectal Cancer Vulnerability To Concomitant Blockade Of Nedd8 And Egfr Pathways

Simple Summary Identification of effective therapies for clinically aggressive, treatment-resistant colorectal cancer (CRC) remains an unmet clinical need. Targeted therapies against the epidermal growth factor receptor (EGFR) signaling axis lead to clinical benefits only in a small fraction of patients due to primary and acquired resistance. We previously showed that the NEDD8 pathway inhibitor pevonedistat induced tumor stabilization in preclinical models of aggressive CRC. Here, through synthetic lethality screenings, we found that pevonedistat could be successfully combined with EGFR pathway-targeted treatments in BRAF-mutant and RAS-RAF wild-type CRCs originally resistant to BRAF and EGFR blockade. We found that combined blockade of NEDD8 and EGFR pathways reverted compensatory feedback loops that reduced the efficacy of single treatments. Our results provide preclinical validation of a promising therapeutic strategy for clinically aggressive CRC resistant to EGFR and BRAF-targeted treatments. Colorectal cancer (CRC) is a heterogeneous disease showing significant variability in clinical aggressiveness. Primary and acquired resistance limits the efficacy of available treatments, and identification of effective drug combinations is needed to further improve patients' outcomes. We previously found that the NEDD8-activating enzyme inhibitor pevonedistat induced tumor stabilization in preclinical models of poorly differentiated, clinically aggressive CRC resistant to available therapies. To identify drugs that can be effectively combined with pevonedistat, we performed a "drop-out" loss-of-function synthetic lethality screening with an shRNA library covering 200 drug-target genes in four different CRC cell lines. Multiple screening hits were found to be involved in the EGFR signaling pathway, suggesting that, rather than inhibition of a specific gene, interference with the EGFR pathway at any level could be effectively leveraged for combination therapies based on pevonedistat. Exploiting both BRAF-mutant and RAS/RAF wild-type CRC models, we validated the therapeutic relevance of our findings by showing that combined blockade of NEDD8 and EGFR pathways led to increased growth arrest and apoptosis both in vitro and in vivo. Pathway modulation analysis showed that compensatory feedback loops induced by single treatments were blunted by the combinations. These results unveil possible therapeutic opportunities in specific CRC clinical settings.