Volumetric compression regulates the phase separation of AXIN and acts as an operational amplifier to bidirectionally modulate Wnt signaling in organoids

Protein phase separation has emerged as a crucial mechanism for spatiotemporal regulation of intracellular processes, yet its potential to integrate and compute diverse extracellular signals is not fully understood. Here, we show a mechano-biochemical circuit that harnesses phase separation to process mechanical and biochemical inputs, modulating cell fate decisions. We demonstrate that volumetric compression bidirectionally regulates canonical Wnt/β-catenin signaling, where the presence of Wnt ligands determines the locations of AXIN phase separation to form either LRP6 signalosomes on the cell membrane or β-catenin destruction complexes in the cytosol, while the mechanical stimulus promotes degree of phase separation to amplify either the positive or negative signal. This circuit enhances healthy intestinal organoid proliferation while suppressing patient-derived colorectal cancer organoid growth, revealing its potential for precise mechanotherapy. Our findings establish phase separation as a critical component in mechanical signal transduction and provide a framework for integrating mechanical and biochemical cues in cellular decision-making. This approach opens avenues for targeted therapies and deepens our understanding of how cells process complex environmental information. The scaffold protein AXIN can promote the formation of two opposing complexes: the Wnt signalosome and a degradation complex that inhibits Wnt. Here, the authors show that mechanical compression and the presence/absence of Wnt ligand are inputs in a circuit that acts as a logic gate to control which complex is formed.