A Novel Suite of Biosensors to Investigate Vinculin Coordination of Collective Cell Migration

Benjamin Johns

Mentors: T. Curtis Shoyer, Brenton Hoffman, Ph.D.

Department of Biomedical Engineering

Collective cell migration (CCM) features a group or chain of linked cells moving together with the same speed in the same direction. This multicellular process is critical in physiological events such as wound healing and morphogenesis. The coordination of forces crucial to CCM is achieved by connecting the actin cytoskeletons of adjacent cells together at dynamic cadherin cell-cell contacts. However, how this mechanical linking regulates CCM at the molecular level remains poorly understood. We hypothesize that the actin-binding protein vinculin mediates these connections to control the speed and coordination of CCM. To test this, we are fusing vinculin to the fluorescent protein mScarletI and creating versions with point mutations that affect specific vinculin protein-protein interactions. With this suite of biosensors, we can visualize vinculin localization and dynamics through fluorescence microscopy and examine vinculin’s effects on CCM speed and coordination through a migration assay. Specifically, we expect that the actin-binding mutant vinculin will alter CCM speed and coordination. Identifying and characterizing these key molecular players of CCM would both greatly advance our understanding of this biological process and possibly provide future targets for therapeutic and tissue engineering purposes.

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