RGD-mediated biomimetic dynamic matrix stiffness signaling enable rapid 3D organoid formation

The stiffness of the extracellular matrix (ECM) is crucial for regulating tissue morphogenesis and cellular behavior. The Arg-Gly-Asp (RGD) motif is a key integrin ligand. However, whether RGD regulates cellular function and tissue remodeling through mechanical stiffness-driven mechanical signaling remains unclear. Here, we develop an easily prepared, ECM-free RGD-functionalized carboxymethylcellulose (CMC) hydrogel. By modulating its mechanical strength, the synergistic effect between RGD-mediated integrin linking and matrix stiffness regulates the formation and expansion of a wide range of tissue-specific organoids through the mechanical transduction pathway of integrin β1/FAK/YAP. Finally, using this method, we develop an acral melanoma organoid model that simulates its development driven by mechanical force and provides a platform for drug screening. These results highlight that RGD-functionalized, stiffness-tunable hydrogel is an excellent biomimetic model for investigating mechanotransduction-driven cell development and disease progression, providing a low-cost strategy for future high-throughput drug screening using organoids.