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.
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.
|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|
|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|
|Liu, Zheng; Liu, Yang; Chang, Yuan et al. (2016) Nanoscale optomechanical actuators for controlling mechanotransduction in living cells. Nat Methods 13:143-6|
|Chang, Yuan; Liu, Zheng; Zhang, Yun et al. (2016) A General Approach for Generating Fluorescent Probes to Visualize Piconewton Forces at the Cell Surface. J Am Chem Soc 138:2901-4|
|Yehl, Kevin; Mugler, Andrew; Vivek, Skanda et al. (2016) High-speed DNA-based rolling motors powered by RNase H. Nat Nanotechnol 11:184-90|
|Stabley, Daniel R; Oh, Thomas; Simon, Sanford M et al. (2015) Real-time fluorescence imaging with 20â€‰nm axial resolution. Nat Commun 6:8307|
|Jurchenko, Carol; Salaita, Khalid S (2015) Lighting Up the Force: Investigating Mechanisms of Mechanotransduction Using Fluorescent Tension Probes. Mol Cell Biol 35:2570-82|
|Boopathy, Archana V; Che, Pao Lin; Somasuntharam, Inthirai et al. (2014) The modulation of cardiac progenitor cell function by hydrogel-dependent Notch1 activation. Biomaterials 35:8103-12|
|Jurchenko, Carol; Chang, Yuan; Narui, Yoshie et al. (2014) Integrin-generated forces lead to streptavidin-biotin unbinding in cellular adhesions. Biophys J 106:1436-46|
|Zheng, Weiwei; Liu, Yang; West, Ana et al. (2014) Quantum dots encapsulated within phospholipid membranes: phase-dependent structure, photostability, and site-selective functionalization. J Am Chem Soc 136:1992-9|
Showing the most recent 10 out of 16 publications