Long-term confinement triggers RIPK3-independent necroptosis in amoeboid cancer cells to shape tumor evolution

Following MLKL activation, L-MAT cells exhibit progressive migration restriction prior to membrane rupture. Notably, MLKL-knockout cells bypass this checkpoint, retain motility, and persist as surviving clones. Spatial transcriptomic analysis of patient tumors reveals that MLKL expression correlates with local PIEZO1 activation and immunogenicity in less aggressive regions, whereas malignant and metastatic clones exhibit reduced MLKL levels.

Our findings establish MLKL as a conditional tumor checkpoint that governs L-MAT cell fate, functioning as an intrinsic barrier against invasive amoeboid cancer cells independent of canonical death-receptor signaling. Counterintuitively, this selective pressure drives tumor evolution by enriching for necroptosis-resistant, aggressive clones. These results uncover an evolutionary bottleneck in cancer progression and identify the PIEZO1-STAT1-MLKL axis as a therapeutic target.