Mark Colasurdo

BME PhD Proposal Presentation

Date: December 14th, 2018

Time: 10:00 AM

Location: CHOA Seminar Room, EBB

Advisor: Andrés J. García (Georgia Institute of Technology)

Committee Members: 

Cheng Zhu (Georgia Institute of Technology)

Jennifer Curtis (Georgia Institute of Technology)

Khalid Salaita (Emory University)

Christian Franck (University of Wisconsin, Madison)

Title: Investigating 3D Vinculin Mechanotransduction using PEG-4MAL Synthetic Hydrogels and 3D Traction Force Microscopy

Abstract/Summary:

Cells interact with the extracellular matrix (ECM) via reciprocal biophysical and biochemical signaling in a process known as mechanotransduction. Mechanotransduction controls multiscale biological processes from cellular proliferation, migration, and differentiation to tissue morphogenesis, pathogenesis, and repair. Cell-matrix interactions are mediated by integrins, focal adhesion (FA) proteins, and the cytoskeleton, which enables cells to sense and transmit mechanical signals from and to their ECM. Vinculin, an FA protein, is considered a primary mediator of mechanotransduction and is responsible for modulating FA assembly, strength, and cell-generated traction forces in response to matrix properties. While vinculin has been widely studied in the past, these studies have occurred using systems that poorly recapitulate the native cellular microenvironment and/or lack control over the biophysical and biochemical properties of the ECM, which are critical aspects of cell-matrix interactions. Therefore, we propose to use a platform combining PEG-4MAL synthetic hydrogels and 3D Traction Force Microscopy (3D TFM), which enables the quantitative analysis of 3D vinculin mechanotransduction within a microenvironment that has both physiologically relevant and precisely controlled biophysical and biochemical properties. Using this platform, we will engineer hydrogels with user-defined matrix elasticity and ligand density to study how vinculin facilitates FA assembly and traction stress generation in 3D with response to varying matrix properties. Then, variants of the vinculin protein will be used to investigate the role of vinculin’s structure and conformation on its ability to sense and transmit force in 3D. By utilizing a platform that has both physiologically relevant and precisely controlled biophysical and biochemical properties, we will be able to gain powerful and unprecedented insights into the role vinculin plays in 3D cellular and molecular mechanotransduction.