AboutThis is the 6th ZOO meeting since 1993. The ZOO meeting series has become a landmark event in the field of cell adhesion and migration due to unique theme selection, high scientific profile with excellent speakers and limited number of attendees ( < 200).
The themes address molecular cell biological principles in the fields of cancer and immunology, with focus on cell-tissue interaction, migration and cell-cell communication. The speakers are leaders in their field and all make extensive use of imaging in their research. Scientific sessions will offer the latest developments in mechanobiology, leukocyte trafficking and cancer cell invasion. Live cell confocal imaging, super resolution imaging, quantitative imaging using innovative approaches and biosensors and computational modelling will be highlighted as part of the presentations.
The meeting will be organized in the Rotterdam Zoo conference facilities.
Photo: Wilbert Boelens.
Anna Akhmanova Utrecht UniversityAnna Akhmanova's group studies cytoskeletal organization and trafficking processes, which contribute to cell polarization, differentiation, vertebrate development and human disease. They are interested in understanding, at a systems level, how different aspects of cell architecture are coordinated.
Anna Akhmanova is a recipient of the ALW Vernieuwingsimpuls VIDI in 2001 and VICI awards in 2007, an ERC Synergy grant in 2013, and Spinoza Prize in 2018.
Jason Cyster University of CaliforniaThe major goals of the Cysterlab are (i) Decipher the guidance cue codes controlling leukocyte migration and interaction events during tissue surveillance and immune responses; (ii) Visualize immune response dynamics using advanced imaging approaches; (iii) Define the selection mechanisms for antibody affinity maturation and that help prevent autoantibody production.
Jason Cyster is currently the recipient of an HHMI Investigator Award and 2 NIH grants.
- Ansel KM, Ngo VN, Hyman PL, Luther SA, Forster R, Sedgwick JD, Browning JL, Lipp M, Cyster JG. 2000. A chemokine-driven positive feedback loop organizes lymphoid follicles. Nature 406:309- 314.
- Matloubian M, Lo CG, Cinamon G, Lesneski MJ, Xu Y, Brinkmann V, Allende ML, Proia RL, Cyster JG. 2004. Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1. Nature 427:355-360.
- Schwab SR, Pereira JP, Matloubian M, Xu Y, Huang Y, Cyster JG. 2005. Lymphocyte sequestration through S1P lyase inhibition and disruption of S1P gradients. Science 309:1735- 1739.
- Allen CD, Okada T, Tang HL, Cyster JG. 2007. Imaging of Germinal Center Selection Events During Affinity Maturation. Science 315:528-531
- Pereira, J.P., Kelly, L.M., Xu, Y. and Cyster, J.G. 2009. EBI2 mediates B cell segregation between the outer and center follicle. Nature 460
- Arnon TI, Xu Y, Lo C, Pham T, An J, Coughlin S, Dorn GW, Cyster JG. 2011. GRK2-dependent S1PR1 desensitization is required for lymphocytes to overcome their attraction to blood. Science 333:1898-1903.
- Muppidi JR, Schmitz R, Green JA, Xiao W, Larsen AB, Braun SE, An J, Xu Y, Rosenwald A, Ott G, Gascoyne RD, Rimsza LM, Campo E, Jaffe ES, Delabie J, Smeland EB, Braziel RM, Tubbs RR, Cook JR, Weisenburger DD, Chan WC, Vaidehi N, Staudt LM, Cyster JG. Loss of signalling via Gα13 in germinal centre B-cell-derived lymphoma. Nature 2014; 516:254-258
Reinhold Förster Hannover Medical SchoolThe working group headed by Prof. Förster sees to the clarification of the cellular and molecular mechanisms forming the basis of the functional organisation of the lymphoid organs. We are particularly interested in clearing up the function of chemokines and chemokine receptors during the migration of immune cells into the secondary lymphoid organs and within these organs.
Reinhold Förster received many grants among SFB grants, DFG grant and an ERC advanced grant in 2013.
Jacco van Rheenen Netherlands Cancer InstituteBy intravital microscopy, the van Rheenen group studies the identity, behavior, and fate of cells that drive tumor initiation, progression, metastasis and the development of therapy resistance.
For his work Jacco van Rheenen received several international awards including the Stem Cells Young Investigator Award, ERC consolidator grant, and the Dr. Josef Steiner Cancer Research Foundation Award.
- Scheele CL, Hannezo E, Zomer A, Langedijk NSM, Simons BD, van Rheenen J, (2017) Identification of mammary stem cells and their dynamics during branching morphogenesis, Nature, 542(7641):313-317.
- Hannezo E, Scheele CLGJ, Moad M, Drogo N, Heer R, Sampogna RV, van Rheenen J#, Simons BD# (2017) A Unifying Theory of Branching Morphogenesis, Cell, Sep 21;171(1):242-255.e27. #co-senior
- Zomer A, Maynard C, Verweij FJ, Kamermans A, Schäfer R, Beerling E, Schiffelers RM, de Wit E, Berenguer J, Ellenbroek SIJ, Wurdinger T, Pegtel DM, van Rheenen J, (2015) In vivo imaging reveals extracellular vesicle-mediated phenocopying of metastatic behavior, Cell, 161(5):1046-1057.
- Ritsma L, Ellenbroek SIJ, Zomer A, Snippert HJ, de Sauvage FJ, Simons BD, Clevers H, van Rheenen J, (2014) Intestinal crypt homeostasis revealed at single stem cell level by in vivo live-imaging, Nature, 507(7492):362-365.
- Philippe Bousso
- Koen van den Dries
- Michael Dustin
- Ben Fabry
- Carsten Grashoff
- Louis Hodgson
- Kairbaan Hodivala-Dilke
- Anna Huttenlocher
- Stephan Huveneers
- Fanny Jaulin
- Matthew Krummel
- Paul Kubes
- Stefan Linder
- Tim Lämmermann
- Danijela Matic Vignjevic
- Sussan Nourshargh
- Steven Pals
- Matthieu Piel
- Martin Schwartz
- Xavier Trepat
Abstracts speakersAnna Akhmanova, Cell Biology, Faculty of Science, Utrecht University, The Netherlands
Control of cell morphology and migration by dynamic microtubules
Microtubules are dynamic polymers that can switch between phases of growth and shortening. This process, termed dynamic instability, is regulated by numerous cellular factors and can be affected by drugs used for cancer therapy. We found that mesenchymal cells embedded in a three-dimensional matrix become immobilized when microtubule growth is attenuated by drugs or due to the lack of specific regulatory factors. Such cells cannot form elongated protrusions and penetrate the matrix, and as a result, cancer cells with perturbed interphase microtubule growth are impaired in forming invasive tumors in mice. To uncover the underlying molecular mechanisms, we have reconstituted some essential properties of microtubule growth regulation in vitro using purified components.
Philippe Bousso,Institut Pasteur , Paris, France
Decoding the mechanisms of tumor immunosurveillance and immunotherapies at the single cell level
Tumors are under the selective pressure of the immune system. While the concept of tumor immunosurveillance is well-established, a number of key questions remains. At what stages of the tumor development does the immune system have the ability to eliminate malignant cells? How and where do cytotoxic effectors such as NK cells or CD8+ T cells restrict tumor progression? How do interactions between immune cells and tumor cells change as the disease progresses? Moreover, the rationale design and optimization of tumor immunotherapies critically require a better understanding of their mechanism of action in vivo. In this presentation, we will discuss how intravital imaging can help gain new insight into mechanisms at play during tumor immunosurveillance and uncover the mode of action of tumor immunotherapies.
Edna Cukierman, Marvin & Concetta Greenberg Pancreatic Cancer Institute, Immune Monitoring Facility, Fox Chase Cancer Center, USA
Oncogenic synapses; stromal regulators of PDAC metabolic support and innate immunosuppression
Expansion of stellate cells, known as cancer associated fibroblasts (CAFs), and extracellular matrix remodeling, is a major culprit for the pancreatic cancer (PC) microenvironment, which restricts nutrients by collapsing blood vessels and fosters immunosuppression. CAFs, PC, and immune cells crosstalk, in this microenvironment, is not fully understood. This seminar sheds light on how CAF-derived matrices trigger a new signaling axis, akin to excitatory synaptic stress, responsible for nutritional CAF support of PC cells and CAF fostering of pro-tumoral immunosuppression. The study identifies novel potential targets for PC, a cancer that is in desperate need of new and more effective therapies.
Jason Cyster, HHMI and University of California, San Francisco, USA
Molecular cues controlling cell migration during the B cell response
Germinal centers (GCs) are important sites of antibody diversification and affinity improvement as well as lymphomagenesis. GC B cells are highly motile and this permits their surveillance for antigen and encounter with helper T cells. Despite this motility, GC B cells are tightly confined to the germinal center structure. Our work has identified a role for Ga13-coupled migration-inhibitory receptors in GC B cell confinement. This talk will give an overview of our studies on cell migration in GCs as well as detailing our latest findings on the molecular cues and mechanisms promoting GC B cell confinement.
Koen van den Dries, Radboudumc, Nijmegen, The Netherlands
Functional alliance between actin, myosin and septins at protrusive podosomes
Podosomes are cytoskeletal structures that orchestrate protrusion, topography sensing and extracellular matrix degradation in cells that need to cross and degrade tissue boundaries. Individual podosomes are micron-size protrusion machineries and they organize into large clusters to allow for mesoscale mechanosensing. By combining advanced super-resolution microscopy approaches with quantitative image analysis we show that podosome clusters are composed of several cytoskeletal submodules consisting of actin, myosin and septins. Together, our results provide a comprehensive molecular basis to understand podosome-mediated cellular protrusiveness and mechanosensation.
Michael Dustin, Kennedy Institute of Rheumatology, University of Oxford, Oxford, UK
How human CD8 memory T cells outcompete naïve counterparts
Memory T cells have multiple features that enable them to be more protective than naïve T cells. We will present data showing that that human memory CD8 T cells have higher synapse propensity, the probability to initiate immunological synapses, compared to naïve counterparts. Higher synapse propensity in human memory CD8 T cells allows them to compete out naïve CD8 T cells from getting recruited to the response. This observation has implications for original antigenic sin and aging of the immune system in humans.
Andy Ewald, Johns Hopkins University School of Medicine
E-cadherin is an invasion suppressor, survival factor, and metastasis promoter across multiple models of breast cancer
E-cadherin (E-cad) mediates adhesion and is considered a metastasis suppressor. However, invasive ductal breast cancers (IDC) retain E-cad in both primary tumors and metastases. To resolve this discrepancy, we tested the requirement for E-cad in three models of IDC. Loss of E-cad increased invasion but also reduced cancer cell proliferation, survival, circulating tumor cell number, tumor cell seeding, and metastasis formation. Transcriptionally, loss of E-cad was associated with increased TGF-ß, reactive oxygen, and apoptosis signaling. Disseminating E-cad-negative cells exhibited nuclear enrichment of SMAD2/3, oxidative stress, and apoptosis. Colony formation of E-cad-negative cells was rescued by treatment with TGF-ß receptor inhibitors, N-acetyl cysteine, or apoptosis inhibitors. These results suggest that E-cad can act as a survival factor during the detachment, systemic dissemination, and seeding phases of metastasis by limiting reactive oxygen-mediated apoptosis.
Ben Fabry, Center for Medical Physics and Technology, Germany
Homing mechanisms of natural killer cells in cancer
Natural killer (NK) cells are important effector cells in the immune response to cancer. NK cells and tumor cells can be co-cultured in 3-dimensional (bio)polymers that serve as a substitute for connective tissue, to investigate mechanisms of NK cell migration, homing and cytotoxicity towards tumor cells. These findings and insights can be utilized for an improved NK cell therapy in patients with solid tumors.
Reinhold Förster, Institute for Immunology, Hanover Medical School, Hanover Germany
CCR7 and its ligands: controlling cells cell migration between and within lymphoid and non-lymphoid organs
A key feature of the immune system is its ability to induce protective immunity against pathogens and to maintain tolerance towards self and innocuous environmental antigens. Accumulating data indicates that, by regulating the migration of immune cells, the chemokine receptor CCR7 essentially contributes to both immunity and tolerance. This receptor is involved in organizing thymic architecture and function, lymph-node homing T cells via high endothelial venules, as well as via afferent lymphatics. Here, I will give an overview on the cellular and molecular mechanisms how CCR7 and its two ligands, CCL19 and CCL21, act as key regulators of the adaptive immune system.
Carsten Grashoff, Max Planck Institute of Biochemistry
Piconewton-sensitive biosensors to investigate adhesion mechanics in cells
To investigate the molecular mechanisms underlying cell adhesion mechanics, we developed a range of single-molecule‒calibrated biosensors that are sensitive to mechanical forces of 3–5, 6–8 and 9–11 piconewton. The tension sensors are characterized by fast folding-unfolding transitions, reversibility and a defined force-response threshold, and can be used to determine molecular forces acting across proteins in cells. Their application to the focal adhesion protein talin and the desmosomal plaque protein desmoplakin reveals intriguing differences in how these molecules modulate intracellular force propagation.
Louis Hodgson, Albert Einstein College of Medicine, New York, USA, Michael F. Price Center, New York, USA
New Windows on Living Cells: Spatiotemporal dynamics of Rho GTPases regulate cancer invasion
P21 Rho family small GTPases are critically important in many disease processes including cancer, inflammatory diseases, and developmental defects. This class of signaling molecules is critical in these diseases by impacting directly: cell polarity, motility and migration through their actions on downstream cytoskeleton/adhesion dynamics. Rho GTPases regulate these processes by coordinating, in both space and time, their activities in response to various environmental cues. However it has been difficult to dissect the coordinated dynamics by conventional approaches. Here, I will introduce several new biosensors specifically targeting different isoforms of Rho GTPases useful for live-cell imaging, based on Förster resonance energy transfer (FRET) that we have developed in our laboratory.
Kairbaan Hodivala-Dilke, Barts Cancer Institute, Queen Mary University of London
Cancer stroma modifications and the control of cancer growth
Our laboratory research aims to unravel the complex intercellular regulation of cancer growth via the inherent cross talk between cells within the tumour microenvironment and malignant cells. Using a combination of in vivo cancer models, stromal specific genetic ablation, proteomics, transcriptomics and metabolomics we dissect the molecular controllers of cancer growth. We exploit these discoveries to develop improved methods of cancer control and chemosensitivity. My talk will detail an example of current findings.
Anna Huttenlocher, University of Wisconsin school of medicine and public health, Madison, USA
Imaging inflammation and its resolution
Motile cells navigate through complex tissues in vivo that include both attractive and repulsive cues. We exploit the optical transparency of zebrafish larvae to image cell migration during wound repair and cancer. We will discuss the heterotypic cell-cell interactions and extracellular cues that mediate inflammation and its resolution.
Stephan Huveneers, AMC Amsterdam
Sensing of forces by (a)symmetric endothelial junctions for angiogenesis
The endothelium receives and responds to mechanical forces through VE-cadherin-based cell-cell junctions. Differences in the cytoskeletal geometries between adjacent cells cause unbalanced forces at the junctional-interface. We previously showed that the F-BAR protein Pacsin2 demarcates force-dependent asymmetric junctions. This introduces a novel paradigm in cadherin mechanotransduction involving plasma membrane curvature as trigger for junctional BAR proteins. Our current work reveals that Pacsin2 recruits trafficking modulators to the junction side of follower cells and thereby steers polarization and drives collective migration. I will demonstrate how pacsin2 propagates directional cues between leader and follower cells and guides endothelial dynamics in angiogenesis.
Fanny Jaulin, Institut de Cancérologie Gustave Roussy, Villejuif, France
Morphogenetic programs in colorectal carcinomas dissemination
Tumor cell dissemination is the first step in metastases formation and the leading cause of cancer-related death. Extensive investigation in a variety of experimental model systems led to the assumption that the fatal progression of carcinomas is associated with the loss of epithelial architecture and apico-basolateral polarity as single cell or undifferentiated cell groups escape the primary tumour. Yet, the mechanisms driving metastatic spread in cancer patients, which depend on primary tumor type, dissemination route, and secondary organ site, remain under-investigated.
Our team has developed translational cell biology approaches to explore colorectal carcinoma (CRC) dissemination from patient specimens. Our results show that CRC mostly undergo collective invasion and maintain their epithelial architecture and robust apico-basolateral polarity as they spread. Depending on their molecular alterations, the CRC cell cohorts either maintain an inward apical pole or revert their topology to expose an outward apical pole, facing patient fluids and tissues. The two topologies are associated with different mode of collective invasion driven by distinct molecular mechanisms. By applying cell biological methods to primary cancer in a clinical setting, we provide an alternative conception of cancer dissemination that has revealed fundamental new cell biology.
Paul Kubes, University of Calgary, Calgary USA
Cavity macrophage: Immunity around organs
While we have known for many years that the peritoneum is filled with macrophage that are easily isolated and studied, only recently have we gained insight as to their function. The large GATA6+ macrophage infiltrate injured visceral organs and help in mediating repair. They can be found in both mice and humans, and in both species significant perturbations cause them to “disappear”. During an infection, these macrophage drop in number and are rapidly replaced by GATA6- small peritoneal macrophage derived from bone marrow monocytes. We observed that the GATA6+ macrophage are particularly susceptible to infection. For example, with S.aureus infection they function as a conduit leading to infection of numerous organs. The peritoneum is not the only cavity in the body, and other cavities include the pericardial and pleural space. In both compartments, GATA6+ macrophage can be found with greater abundance in humans. During an ischemic injury to the heart, the pericardial macrophage appear to regulate the degree of fibrosis that can be found in the non-injured part of the heart. In conclusion, the GATA6+ cavity macrophage are located in numerous compartments and can mediate tissue repair but also can be hijacked by various infectious agents.
T. Lämmermann, Max Planck Institute of Immunobiology and Epigenetics (MPI-IE), Freiburg, Germany
Insterstitial migration modes of tissue-resident immune cells
Most cells of the immune system stand out against other body cell types owing to their extremely fast tissue movement and flexibility to invade diverse organs. Studies on fast-migrating leukocytes
(neutrophils, lymphocytes, dendritic cells) shaped the current view that amoeboid leukocyte migration does not require integrin-mediated adhesions within tissues and is largely independent from the molecular composition of the environment. Instead, these leukocyte subsets almost exclusively rely on cell shape changes driven by the actomyosin cytoskeleton during their migration outside the vasculature. Our current work sets focus on the tissue dynamics and 3D movements of tissue-resident, slow-moving leukocyte subsets and how their migration modes relate to the current paradigm of integrin-independent interstitial leukocyte migration.
Stefan Linder, University Medical Center Eppendorf, Hamburg, Germany
Regulation and analysis of actomyosin-dependent contractility at macrophage podosomes
Podosomes are multifunctional adhesion and invasion structures of macrophages. They are also able to work as mechanosensory organelles, based on their specialized architecture and the local regulation of actomyosin-dependent contractility. Especially the podosome cap, a more recently described substructure, has emerged as a regulatory module of podosomal contractility, thus regulating oscillatory protrusion and mechansensing of these organelles. This talk will give an update on podosome substructure, present new findings on the regulation of cap-associated actomyosin contractility and introduce new tools for the analysis of podosome architecture and function.
Danijela Matic Vignjevic, Institut Curie, Paris, France
The dark side of fibroblast force
Tumor microenvironment plays an important role in the tumor progression. It is made of extracellular matrix (ECM), blood vessels, immune cells and cancer-associated fibroblasts (CAFs). Besides biochemical signals, mechanical forces from microenvironment also play a role in tumor progression. CAFs have enhanced contractility and capacity to synthesize, deposit and crosslink ECM making stroma stiffer. Thus, by accumulating around the tumor, they could provide a physical barrier constraining tumor expansion. However, it has been shown that by exerting mechanical forces on the ECM, CAFs also enhance tumor invasion. These antagonistic roles of forces produced by CAFs in tumor progression will be discussed.
Sussan Nourshargh, William Harvey Research Institute, Barts and The London Medical School, Queen Mary University of London
Neutrophil breaching of venular walls: Mode, dynamics & mechanisms
Neutrophil migration into tissues forms a fundamental arm of innate immunity but is also a key instigator of numerous inflammatory disorders. Despite our increased knowledge of this process, there remain many open questions. Our group aims to decipher the mechanisms through which neutrophils breach venular walls, applying high resolution confocal intravital microscopy to analysis of neutrophil-endothelial cell and neutrophil-pericyte interactions in vivo. Our work has shed light on the mode and dynamics of such fundamental responses and identified key molecular players. Furthermore, we have discovered numerous disrupted modes of neutrophil transendothelial cell migration (TEM) that could have pathological implications. Principal concepts will be discussed.
Steven T. Pals, Department of Pathology, Amsterdam University Medical Centers, and Lymphoma and Myeloma Center Amsterdam – LYMMCARE, Amsterdam, The Netherlands.
Provoking homelessness as strategy to treat lymphoma
Tumor cells are greatly influenced by, or even fully addicted to signals from the microenvironment. As targets for therapeutic intervention these signals are potentially equally important as those driven by mutated oncogenes. In normal and malignant lymphocytes, signals emanating from the antigen- and chemokine-receptors play a central role in the control of interaction with the microenvironment by regulating the activity of integrin adhesion receptors. Apart from controlling the temporal and spatial interactions of lymphocytes with their microenvironment, which provides indispensable growth- and survival factors, integrins also emit signals that directly promote cell growth and survival. These activities of integrins play a quintessential role in regulating lymphocyte fitness and prevent cell death by a specific type of programmed cell death, termed anoikis (homelessness). As will be discussed, data from our own and other laboratories advocate targeting of the molecular pathways controlling integrin-mediated adhesion, homing, and retention of malignant lymphocytes as a strategy to treat lymphoma.
Matthieu Piel, Science Institut Curie, Paris, France
Cellular branching and nuclear deformation during fast immune cell migration through dense tissues
When immune cells migrate through tissues or, in vitro, through denses matrices such as collagen gels, they undergo large deformations because they do not degrade the matrix as they move. The two main factors that have been proposed to limit their speed, and potentially lower it compared to their intrinsic motile capicity, are the induction of cellular branching by obstacles such as collagen fibers, and the deformation of their nucleus. I will discuss which phenomenon is actually dominating depending on the migration environment and the presence of chemokine gradients. I will also present specific mechanisms that immune cells possess to choose between branches and deform their nucleus efficiently, thus reducing these trapping factors when they migrate through dense matrices.
Jacco van Rheenen, Department of Molecular Pathology, Oncode Institute, Netherlands Cancer Institute Amsterdam, the Netherlands
Intravital microscopy revealed cancer stem cell plasticity to be important for metastasis
Although histological techniques have provided important information on stem cells and cancer, they draw static images of dynamic processes. To study dynamic processes, we have developed intravital microscropy techniques to monitor these cells in real time in living animals. We have used these techniques to study the identity and (migratory) behavior of (cancer) stem cells during pubertal mammary morphogenesis, during tumor growth, and during the metastatic process. Our IVM experiments illustrate that cellular properties and fate of (cancer) stem cells are highly dynamic, and that stemness is a state as opposed to an intrinsic property of a cell.
Martin Schwartz, Yale Cardiovascular Research Center
Force transmission and mechanotransduction through integrins
Xavier Trepat, ICREA @ Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain, Barcelona Institute for Science and Technology (BIST), Barcelona, Spain
Mechanobiology of epithelial migration, growth and folding
Biological processes such as morphogenesis, tissue regeneration, and cancer invasion are driven by collective migration, division, and folding of epithelial tissues. Each of these functions is tightly regulated by mechanochemical networks and ultimately driven by physical forces. I will present maps of cell-cell and cell-extracellular matrix (ECM) forces during cell migration and division in a variety of epithelial models, from the expanding MDCK cluster to the regenerating zebrafish epicardium. These maps revealed that migration and division in growing tissues are jointly regulated. I will also present direct measurements of epithelial traction, tension, and luminal pressure in three-dimensional epithelia of controlled size and shape. By examining epithelial tension over time-scales of hours and for nominal strains reaching 1000%, we establish a remarkable degree of tensional homeostasis mediated by superelastic behavior.