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

ABSTRACT: The mesenchymal–amoeboid transition (MAT) confers enhanced migratory capacity and therapy resistance to cancer cells. To disseminate, MAT cancer cells squeeze through compacted intra-tumor spaces, breach matrix barriers, and infiltrate into the vasculature—a journey defined by sustained mechanical pressure. Yet how prolonged MAT under such pressure influences cell fate remains poorly understood. Here, we introduce the concept of “Long-term MAT” (L-MAT)—referring to cells with sustained MAT status under pressure—and demonstrate that L-MAT in non-adhesive three-dimensional microenvironments triggers a previously unrecognized RIPK3-independent necroptotic pathway. Mechanistically, the PIEZO1-STAT1 axis mediates transcriptional upregulation of MLKL, and elevated MLKL accumulates on the membrane even without detectable RIPK3 expression, driving necroptosis in cells traditionally considered necroptosis-resistant in 2D petri dish. 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.

ORGANISM(S): Mus musculus Homo sapiens

PROVIDER: GSE328536 | GEO | 2026/09/05

REPOSITORIES: GEO