The Notch receptors coordinate cell to cell communication, cell growth, and determine cell fates. Malfunctions in Notch signaling have been linked to a range of diseases including cancer, which has spurred interest in understanding its molecular mechanisms of activation. The central hypothesis of the proposed work is that physical aspects of the Notch ligand-receptor interaction, mechanotransduction, and spatial arrangement, actively provide a molecular mechanism for signal regulation. The long-term goal of this proposal is to develop a complete understanding of how spatio-mechanical inputs can exert regulatory control over biochemical signaling processes. We seek a fundamental understanding of how the extracellular environment influences a cell's intracellular chemical signaling. To achieve this goal, we propose a highly multidisciplinary, hybrid physical and biological approach aimed at deconstructing how the Notch receptor cleavage is sensitive to clustering, mechanics, and spatial arrangement. These questions cannot be addressed unless a new approach is introduced to manipulate and to investigate Notch in individual living cells. We will employ surface-based activation of Notch to recapitulate its innate two-dimensional geometry with the goal of addressing long-standing questions regarding the role of spatial and temporal inputs and stochastic noise in triggering the signaling pathway. Preliminary data indicates that the synthetic lipid membrane platform provides for a more physiologically accurate approach to activate the Notch pathway in mammalian cell lines. A newly developed fluorescence force sensor will allow the direct measurement of the association between mechanical strain and protease activity. Microscopy-based single cell analysis of the transcriptional program will be used to measure ligand-induced activation. These experiments will yield a quantitative description of the pathway and may help in understanding the role of molecular Notch deregulations in human cancers.

Public Health Relevance

The Notch pathway is universally employed between animal cells to control differentiation, growth, and development. Malfunctions in processing and relaying Notch signals are responsible for a range of human disorders and cancers. The goal of this proposal is to better understand Notch signaling by developing novel approaches to trigger the pathway and characterize its response functions in mammalian cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM097399-03
Application #
8643259
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Nie, Zhongzhen
Project Start
2012-04-01
Project End
2017-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
3
Fiscal Year
2014
Total Cost
$291,029
Indirect Cost
$82,921
Name
Emory University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Petree, Jessica R; Yehl, Kevin; Galior, Kornelia et al. (2018) Site-Selective RNA Splicing Nanozyme: DNAzyme and RtcB Conjugates on a Gold Nanoparticle. ACS Chem Biol 13:215-224
Su, Hanquan; Liu, Zheng; Liu, Yang et al. (2018) Light-Responsive Polymer Particles as Force Clamps for the Mechanical Unfolding of Target Molecules. Nano Lett 18:2630-2636
Glazier, Roxanne; Salaita, Khalid (2017) Supported lipid bilayer platforms to probe cell mechanobiology. Biochim Biophys Acta Biomembr 1859:1465-1482
Liu, Zheng; Liu, Yang; Chang, Yuan et al. (2016) Nanoscale optomechanical actuators for controlling mechanotransduction in living cells. Nat Methods 13:143-6
Somasuntharam, Inthirai; Yehl, Kevin; Carroll, Sheridan L et al. (2016) Knockdown of TNF-? by DNAzyme gold nanoparticles as an anti-inflammatory therapy for myocardial infarction. Biomaterials 83:12-22
Liu, Yang; Blanchfield, Lori; Ma, Victor Pui-Yan et al. (2016) DNA-based nanoparticle tension sensors reveal that T-cell receptors transmit defined pN forces to their antigens for enhanced fidelity. Proc Natl Acad Sci U S A 113:5610-5
Yehl, Kevin; Mugler, Andrew; Vivek, Skanda et al. (2016) High-speed DNA-based rolling motors powered by RNase H. Nat Nanotechnol 11:184-90
Ma, Victor Pui-Yan; Liu, Yang; Blanchfield, Lori et al. (2016) Ratiometric Tension Probes for Mapping Receptor Forces and Clustering at Intermembrane Junctions. Nano Lett 16:4552-9
Ma, Victor Pui-Yan; Liu, Yang; Yehl, Kevin et al. (2016) Mechanically Induced Catalytic Amplification Reaction for Readout of Receptor-Mediated Cellular Forces. Angew Chem Int Ed Engl 55:5488-92
Galior, Kornelia; Liu, Yang; Yehl, Kevin et al. (2016) Titin-Based Nanoparticle Tension Sensors Map High-Magnitude Integrin Forces within Focal Adhesions. Nano Lett 16:341-8

Showing the most recent 10 out of 22 publications