Cell polarization and directed migration are fundamental processes required for human cells to mediate immune responses, wound healing, angiogenesis, development, and cancer metastasis. While much work has been done to identify and characterize regulatory proteins involved in these processes, the large number of critical proteins and second messengers has left the questions open of how cells induce polarization, drive migration and trigger chemotaxis towards a source of chemoattractant. The goal of the proposed work is to answer these questions by employing a microscopy based quantitative analysis of the regulatory control systems in endothelial cells and neutrophils. Understanding how these regulatory systems are build will likely reveal critical control points that have potentil as therapeutic targets. Our specific strategy is based on cell lines that we developed expressing FRET- and translocation- based fluorescent biosensors for three different core second messengers, PIP3, Ca2+ and diacylgycerol as well as for four core small GTPases, Ras, RhoA, Rac and Cdc42. We have also developed rapid perturbation strategies, based on synthetic expressed DNA constructs that allow us to use small molecules to rapidly activate each of these core regulators of polarization and identify and characterize the relevant local feedback loops in the two cell models. This already led to the identification of important proteins that connect these core regulators to each other. Furthermore, by being able to track cells and monitor local signaling activities as they migrate and turn towards sources of chemoattractant, we have been able to identify an unexpected central role of Cdc42 in the polarization and directed migration of both cells. The fundamental nature of our experiments on polarization and directed cell migration will make our studies relevant to understand these processes in a wide range of cell types and organisms. Our studies will also provide important tools for other researchers to use for studies of polarization and migration.
Cell polarization and directed migration play critical roles in human immune responses, wound healing, angiogenesis, development, and cancer metastasis. Our cell-based fluorescence microscopy approach will provide systems level molecular and mechanistic insights into the internal control networks that ultimately allow human endothelial cells to form new vasculature and enable human neutrophils to chemotax. The fundamental nature of our experiments on polarization and directed cell migration will make our studies relevant to understand these processes in a wide range of cell types and organisms.
|Cappell, Steven D; Mark, Kevin G; Garbett, Damien et al. (2018) EMI1 switches from being a substrate to an inhibitor of APC/CCDH1 to start the cell cycle. Nature 558:313-317|
|Yang, Hee Won; Chung, Mingyu; Kudo, Takamasa et al. (2017) Competing memories of mitogen and p53 signalling control cell-cycle entry. Nature 549:404-408|
|Winans, Amy M; Collins, Sean R; Meyer, Tobias (2016) Waves of actin and microtubule polymerization drive microtubule-based transport and neurite growth before single axon formation. Elife 5:e12387|
|Nguyen, Trang Thi Thu; Park, Wei Sun; Park, Byung Ouk et al. (2016) PLEKHG3 enhances polarized cell migration by activating actin filaments at the cell front. Proc Natl Acad Sci U S A 113:10091-6|
|Miyazaki, Yusuke; Mizumoto, Kota; Dey, Gautam et al. (2016) A method to rapidly create protein aggregates in living cells. Nat Commun 7:11689|
|Yang, Hee Won; Collins, Sean R; Meyer, Tobias (2016) Locally excitable Cdc42 signals steer cells during chemotaxis. Nat Cell Biol 18:191-201|
|Hayer, Arnold; Shao, Lin; Chung, Mingyu et al. (2016) Engulfed cadherin fingers are polarized junctional structures between collectively migrating endothelial cells. Nat Cell Biol 18:1311-1323|
|Dey, Gautam; Jaimovich, Ariel; Collins, Sean R et al. (2015) Systematic Discovery of Human Gene Function and Principles of Modular Organization through Phylogenetic Profiling. Cell Rep :|
|Collins, Sean R; Yang, Hee Won; Bonger, Kimberly M et al. (2015) Using light to shape chemical gradients for parallel and automated analysis of chemotaxis. Mol Syst Biol 11:804|
|Ge, Xuecai; Milenkovic, Ljiljana; Suyama, Kaye et al. (2015) Phosphodiesterase 4D acts downstream of Neuropilin to control Hedgehog signal transduction and the growth of medulloblastoma. Elife 4:|
Showing the most recent 10 out of 30 publications