A key finding: we observed a loss of rotational migration in pancreatic cancer organoids compared to normal ones, suggesting its potential role in maintaining normal tissue function. We hope this versatile model enables future studies with human primary cells! #CancerResearch

Furthermore, inhibition of actin polymerization (Latrunculin A) and actin branching (CK-666) disrupted rotation and cryptic lamellipodia formation, highlighting the importance of actin organization.

Our inhibition studies using Blebbistatin (myosin II inhibitor) and Y-27632 (ROCK inhibitor) confirmed that myosin activity is essential, as treated organoids showed significant slowdown.

Mechanistically, we found prominent actin stress fibers and focal adhesion clusters at the basal side, interacting with the ECM (Matrigel). We also found enrichment of activated myosin (pMYL9) on these fibers, suggesting actomyosin contractility plays a key role.

By developing a 3D single-cell tracking method, we were able to quantify the rotation velocity, finding a stable average of ~15–20 µm/hr. Our data shows this motion is an inherent feature of these 3D structures.

Movie for the evolution of the cell orientation in zebrafish CE.