Kamboj, S.; Boche, A.; Moret, A.; Wang, Z.; Aime, C.; Agniel, R.; Leroy-Dudal, J.; Carreiras, F.; Gallet, O.; Royle, S. J.; Lambert, A.

Integrins are heterodimeric cell surface receptors that govern cell-cell interactions, which in turn can influence multiscale processes: cell migration, extracellular matrix remodeling and tissue formation. These processes occur over timescales which range from milliseconds to days. While various strategies exist to study integrin function across biological scales from cell to tissue, they are often chronic and fail to target specific cell-cell interactions acutely. We engineered cells to rapidly alter cell behavior by downregulating the surface population of 5{beta}1 integrins through hot-wired clathrin-mediated endocytosis. This method allows for inducible, specific internalization of 5{beta}1 integrins, achieving acute downregulation across various cell lines in 5-30 minutes. We show that induced internalization of 5{beta}1 decreases the cell area, causes uptake of extracellular fibronectin, and decreases the rate of tumor spheroid compaction. This targeted control of multiscale processes by rapid downregulation of this important class of cell surface receptors demonstrates that hot-wired endocytosis is a useful tool to acutely modulate cell biology.