Network Crosstalk Dynamically Changes during Neutrophil Polarization

How complex signaling networks shape highly coordinated, multistep cellular responses is poorly understood. Here, we made use of a network-perturbation approach to investigate causal influences, or "crosstalk," among signaling modules involved in the cytoskeletal response of neutrophils to chemoattractant. We quantified the intensity and polarity of cytoskeletal marker proteins over time to characterize stereotyped cellular responses. Analyzing the effects of network disruptions revealed that, not only does crosstalk evolve rapidly during polarization, but also that intensity and polarity responses are influenced by different patterns of crosstalk. Interestingly, persistent crosstalk is arranged in a surprisingly simple circuit: a linear cascade from front to back to microtubules influences intensities, and a feed-forward network in the reverse direction influences polarity. Our approach provided a rational strategy for decomposing a complex, dynamically evolving signaling system and revealed evolving paths of causal influence that shape the neutrophil polarization response.

[1]  A. Oudenaarden,et al.  A Systems-Level Analysis of Perfect Adaptation in Yeast Osmoregulation , 2009, Cell.

[2]  Garrett M. Odell,et al.  An agent-based model contrasts opposite effects of dynamic and stable microtubules on cleavage furrow positioning , 2008, The Journal of cell biology.

[3]  G. Bokoch,et al.  Nucleotide exchange factor GEF-H1 mediates cross-talk between microtubules and the actin cytoskeleton , 2002, Nature Cell Biology.

[4]  J. Gallin,et al.  Effects of taxol on human neutrophils. , 1982, Journal of immunology.

[5]  H. Bourne,et al.  PDZRhoGEF and myosin II localize RhoA activity to the back of polarizing neutrophil-like cells , 2007, The Journal of cell biology.

[6]  James Watras,et al.  Bell-shaped calcium-response curves of lns(l,4,5)P3- and calcium-gated channels from endoplasmic reticulum of cerebellum , 1991, Nature.

[7]  Madhusudan Natarajan,et al.  A global analysis of cross-talk in a mammalian cellular signalling network , 2006, Nature Cell Biology.

[8]  T. Mak,et al.  Polarization of Chemoattractant Receptor Signaling During Neutrophil Chemotaxis , 2022 .

[9]  Reinhart Heinrich,et al.  The Roles of APC and Axin Derived from Experimental and Theoretical Analysis of the Wnt Pathway , 2003, PLoS biology.

[10]  Peter Donnelly,et al.  Superfamilies of Evolved and Designed Networks , 2004 .

[11]  Olivier Pertz,et al.  Neutrophil polarization: spatiotemporal dynamics of RhoA activity support a self-organizing mechanism. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[12]  S. Gygi,et al.  Hem-1 Complexes Are Essential for Rac Activation, Actin Polymerization, and Myosin Regulation during Neutrophil Chemotaxis , 2006, PLoS biology.

[13]  Zachary D. Smith,et al.  Unbiased Reconstruction of a Mammalian Transcriptional Network Mediating Pathogen Responses , 2009 .

[14]  R. Germain,et al.  Navigating the network: signaling cross-talk in hematopoietic cells , 2009, Nature Immunology.

[15]  M. Gerstein,et al.  Genomic analysis of regulatory network dynamics reveals large topological changes , 2004, Nature.

[16]  A. Bøyum,et al.  Isolation of mononuclear cells and granulocytes from human blood. , 1968 .

[17]  V. Niggli.  Microtubule-disruption-induced and chemotactic-peptide-induced migration of human neutrophils: implications for differential sets of signalling pathways , 2003, Journal of Cell Science.

[18]  K. Sachs,et al.  Causal Protein-Signaling Networks Derived from Multiparameter Single-Cell Data , 2005, Science.

[19]  Daniel Kalman,et al.  Rac and Cdc42 play distinct roles in regulating PI(3,4,5)P3 and polarity during neutrophil chemotaxis , 2003, The Journal of cell biology.

[20]  I. Nemenman,et al.  Information Transduction Capacity of Noisy Biochemical Signaling Networks , 2011, Science.

[21]  Roger Patient,et al.  Transcriptional networks regulating hematopoietic cell fate decisions , 2007, Current opinion in hematology.

[22]  V. Sourjik,et al.  Dynamic map of protein interactions in the Escherichia coli chemotaxis pathway , 2009, Molecular systems biology.

[23]  R. Yu,et al.  Fus3 generates negative feedback that improves information transmission in yeast pheromone response , 2008, Nature.

[24]  F. Maxfield,et al.  Microtubule asymmetry during neutrophil polarization and migration. , 2002, Molecular biology of the cell.

[25]  D. Pe’er,et al.  Principles and Strategies for Developing Network Models in Cancer , 2011, Cell.

[26]  Marc W Kirschner,et al.  An Actin-Based Wave Generator Organizes Cell Motility , 2007, PLoS biology.

[27]  Eugenio Marco,et al.  Endocytosis Optimizes the Dynamic Localization of Membrane Proteins that Regulate Cortical Polarity , 2007, Cell.

[28]  S. Zigmond,et al.  Sensory adaptation of leukocytes to chemotactic peptides , 1979, The Journal of cell biology.

[29]  M. Schliwa,et al.  Centrosome splitting in neutrophils: An unusual phenomenon related to cell activation and motility , 1982, Cell.

[30]  S. M. Goldin,et al.  Calcium as a coagonist of inositol 1,4,5-trisphosphate-induced calcium release. , 1991, Science.

[31]  W. Rappel,et al.  Directional sensing in eukaryotic chemotaxis: a balanced inactivation model. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[32]  P. Bork,et al.  Dynamic Complex Formation During the Yeast Cell Cycle , 2005, Science.

[33]  S L Diamond,et al.  Neutrophil-bead collision assay: pharmacologically induced changes in membrane mechanics regulate the PSGL-1/P-selectin adhesion lifetime. , 2005, Biophysical journal.

[34]  Jingsong Xu,et al.  Divergent Signals and Cytoskeletal Assemblies Regulate Self-Organizing Polarity in Neutrophils , 2003, Cell.

[35]  G. Feng,et al.  The protein tyrosine phosphatase Shp-2 regulates RhoA activity , 2000, Current Biology.

[36]  Uri Alon,et al.  Using a Quantitative Blueprint to Reprogram the Dynamics of the Flagella Gene Network , 2004, Cell.

[37]  Yanqin Wang,et al.  On identifying information from image-based spatial polarity phenotypes in neutrophils , 2010, 2010 IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[38]  D. Lauffenburger,et al.  The Response of Human Epithelial Cells to TNF Involves an Inducible Autocrine Cascade , 2006, Cell.

[39]  Marc W. Kirschner,et al.  A PtdInsP3- and Rho GTPase-mediated positive feedback loop regulates neutrophil polarity , 2002, Nature Cell Biology.

[40]  Benjamin Geiger,et al.  How do microtubules guide migrating cells? , 2002, Nature Reviews Molecular Cell Biology.

[41]  Nikita Vladimirov,et al.  Thermal Robustness of Signaling in Bacterial Chemotaxis , 2011, Cell.

[42]  G. Danuser,et al.  Protein Kinase A Governs a RhoA-RhoGDI Protrusion-Retraction Pacemaker in Migrating Cells , 2011, Nature Cell Biology.

[43]  A. Böyum,et al.  Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. , 1968, Scandinavian journal of clinical and laboratory investigation. Supplementum.

[44]  S. Zigmond,et al.  Cell polarity: an examination of its behavioral expression and its consequences for polymorphonuclear leukocyte chemotaxis , 1981, The Journal of cell biology.

[45]  M. Glogauer,et al.  Rac1 links leading edge and uropod events through Rho and myosin activation during chemotaxis. , 2006, Blood.

[46]  J. Xu,et al.  Neutrophil microtubules suppress polarity and enhance directional migration. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Jason M Haugh,et al.  Mechanisms of Gradient Sensing and Chemotaxis: Conserved Pathways, Diverse Regulation , 2006, Cell cycle.

[48]  Lani F. Wu,et al.  Multidimensional Drug Profiling By Automated Microscopy , 2004, Science.

[49]  Kevan M. Shokat,et al.  To stabilize neutrophil polarity, PIP3 and Cdc42 augment RhoA activity at the back as well as signals at the front , 2006, The Journal of cell biology.