It is now widely acknowledged that cell behavior is highly sensitive to mechanical crosstalk with the extracellular matrix (ECM). While many powerful methods have been developed to control this communication through manipulation of the ECM, there are few tools available for the direct, cell-intrinsic control of cellular mechanotransductive signaling. In this proposal we advance and apply a genetic strategy we recently developed in which we control cell-ECM mechanical signaling through inducible expression of mechanotransductive genes. We have shown that this method enables graded and dynamic control of cortical stiffness, traction force generation, cell migration speed, and ECM remodeling. We have also shown that this approach vastly outperforms traditional pharmacologic strategies in terms of dose-response relationship, target availability, toxicity, and duration of action. We now propose to develop a second generation of this strategy and leverage it to address two unmet needs in the field of cell mechanobiology: First, we will place two genes under the control of promoters that can be induced or suppressed by two different small molecules, thereby enabling orthogonal control over two mechanotransductive genes. We will use this capability to construct a """"""""phase diagram"""""""" of cell mechanical properties that quantitatively maps how the myosin activators Rho- associated kinase and myosin light chain kinase contribute to mechanobiological phenotype. Second, we will apply this strategy to quantitatively control how ECM mechanical properties regulate two important cell behaviors: cell motility speed and neural stem cell neurogenesis. In successful, this will enable us to decouple mechanically-triggered cell behaviors from the inputs themselves, thus potentially offering a way to """"""""rewire"""""""" cell-matrix crosstalk to achieve desired phenotypic endpoints in arbitrarily specified microenvironments. This could offer a new and very powerful way to engineer cell behavior at cell-material interfaces in vitro and in vivo. Taken together, these studies will provide key proof of-principle for this approach as a tool for both quantitative cell biological discovery and cell ad tissue engineering/regenerative medicine applications.

Public Health Relevance

Cell behavior is exquisitely sensitive to the exchange of mechanical signals between cells and the extracellular matrix (ECM). Efforts to leverage this relationship to engineer cell behavior have largely been limited to indirect manipulation of this signaling through control of ECM properties. In this proposal we develop and apply a novel genetic approach to this problem involving inducible expression of mechanotransductive genes, which we envision will serve as a powerful tool for scientific discovery and for controlling cell behavior at biointerfaces in tissue engineering and regenerative medicine applications.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21EB016359-01A1
Application #
8584161
Study Section
Biomaterials and Biointerfaces Study Section (BMBI)
Program Officer
Hunziker, Rosemarie
Project Start
2013-07-01
Project End
2015-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
1
Fiscal Year
2013
Total Cost
$234,750
Indirect Cost
$84,750
Name
University of California Berkeley
Department
Miscellaneous
Type
Organized Research Units
DUNS #
124726725
City
Berkeley
State
CA
Country
United States
Zip Code
94704
Kassianidou, Elena; Brand, Christoph A; Schwarz, Ulrich S et al. (2017) Geometry and network connectivity govern the mechanics of stress fibers. Proc Natl Acad Sci U S A 114:2622-2627
Shyer, Amy E; Rodrigues, Alan R; Schroeder, Grant G et al. (2017) Emergent cellular self-organization and mechanosensation initiate follicle pattern in the avian skin. Science 357:811-815
Kassianidou, Elena; Hughes, Jasmine H; Kumar, Sanjay (2017) Activation of ROCK and MLCK tunes regional stress fiber formation and mechanics via preferential myosin light chain phosphorylation. Mol Biol Cell 28:3832-3843
Chen, Joseph; Kumar, Sanjay (2017) Biophysical Regulation of Cancer Stem/Initiating Cells: Implications for Disease Mechanisms and Translation. Curr Opin Biomed Eng 1:87-95
Lee, Jessica P; Kassianidou, Elena; MacDonald, James I et al. (2016) N-terminal specific conjugation of extracellular matrix proteins to 2-pyridinecarboxaldehyde functionalized polyacrylamide hydrogels. Biomaterials 102:268-76
Guillou, Lionel; Dahl, Joanna B; Lin, Jung-Ming G et al. (2016) Measuring Cell Viscoelastic Properties Using a Microfluidic Extensional Flow Device. Biophys J 111:2039-2050
Hughes, Jasmine Hannah; Kumar, Sanjay (2016) Synthetic mechanobiology: engineering cellular force generation and signaling. Curr Opin Biotechnol 40:82-89
Gould, Russell A; Yalcin, Huseyin C; MacKay, Joanna L et al. (2016) Cyclic Mechanical Loading Is Essential for Rac1-Mediated Elongation and Remodeling of the Embryonic Mitral Valve. Curr Biol 26:27-37
Chang, Ching-Wei; Kumar, Sanjay (2015) Differential Contributions of Nonmuscle Myosin II Isoforms and Functional Domains to Stress Fiber Mechanics. Sci Rep 5:13736
Rape, Andrew D; Zibinsky, Mikhail; Murthy, Niren et al. (2015) A synthetic hydrogel for the high-throughput study of cell-ECM interactions. Nat Commun 6:8129

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