Chemotaxis occurs during a number of physiological events including angiogenesis, embryonic development, wound healing, immune defense, and the establishment of neuronal circuits. Accordingly, eukaryotic chemotaxis has been a topic of key interest in cell biology and pathophysiology. The objective of our research is to explore a long-standing conundrum in the field: How do cells "break symmetry" to initiate cell migration? To address this question, we will focus on two signaling modules that remain uncharacterized: "Feedback" and "Crosstalk". Experimental dissection of these modules has proven inherently challenging, because their signaling operation in cells is local (sub-cellular) and rapid (second-to-minute) on top of their intimate intra- and inter-module relationships. The experimental perturbation of component molecules thus must be restricted to precise spatial domains and be faster than the signaling events. However, most tools used to probe signaling events are generally slow (minute- to-day) and global (supra-cellular) in their effects, limiting their usefulness. We previously developed a new generation of molecular tools that allows for Rapid, Inducible and Specific Perturbation (RISP) of various proteins in living cells. In order to decipher the kinetics and dynamics of molecular networks within the modules with high spatio-temporal resolution, we employ two different approaches: 1. operating the RISP in microfluidic device and 2. improving the present RISP by using synthetic photo- chemistry knowledge. These experimentations will allow us to determine whether and how the elementary signaling modules are integrated to orchestrate an intricate symmetry breaking process. A better understanding of the chemotaxis promises therapeutic advancements for cell migration-related diseases. Our unique approach for probing cellular dynamics will also provide a general-and-powerful methodology that has the potential to significantly extend conventional methods such as RNA interference and pharmacology.

Public Health Relevance

Due to its significant involvement in biological processes, cell migration contributes to disease progression in cancer and arthritis, while its impairment leads to anomalous tissue development or regeneration. Our unique perturbation approach can be a powerful strategy for not only dissecting the molecular mechanisms, but also interfering with these cell migration-related diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM092930-03
Application #
8282785
Study Section
Membrane Biology and Protein Processing (MBPP)
Program Officer
Deatherage, James F
Project Start
2010-07-15
Project End
2015-05-31
Budget Start
2012-06-01
Budget End
2013-05-31
Support Year
3
Fiscal Year
2012
Total Cost
$308,484
Indirect Cost
$120,384
Name
Johns Hopkins University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
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