A. C. Lu, L. Moeller, S. Moore, K. Ho, V. Tobin, A. Qiang, E. Zhang, Z. Li, M. Wang, S. Xia, M. W. Budde, H. Larson, A. Ahmed Diaz, B. Gu, J. M. Linton, L. Klock, M. J. Flynn, Q. Igomu, X. J. Gao, D. J. Siegwart, H. Zhu, M. B. Elowitz
A central challenge in cancer therapy is selectively killing cancer cells while minimizing resistance. Here, we engineer modular protease-based protein circuits that sense mutant RAS, the most frequently mutated oncogene in cancer, and conditionally activate cell death. Delivered transiently as mRNA in lipid nanoparticles (LNPs), circuits selectively eliminated RAS-mutant cancer cells in culture and suppressed aggressive, multifocal RAS-driven liver tumors. Compared to RAS inhibitors, circuits killed cancer cells independently of oncogene addiction, achieved potent cytotoxicity rather than cytostasis, and functioned at low RAS occupancy through catalysis rather than stoichiometric inhibition. Critically, circuits acquired minimal to no resistance under prolonged selection, while remaining potent against prevalent drug-resistance mechanisms including RAS amplification and bypass signaling. Together, these results establish design principles for engineering therapeutic protein circuits and highlight their potential to overcome longstanding limitations of existing therapeutic modalities.