Cell Migration Lecture Notes (Fall Semester 2024) PDF

Dynamics of cellular contacts: Cell-cell contacts and cell motility Introduction – Fall Semester 2024 Cell-cell junctions and cell migration at a glance Coordinator: Britta Engelhardt: [email protected] Introduction: Cell-cell junctions and cell migration at a glance > Part I G...

Dynamics of cellular contacts: Cell-cell contacts and cell motility Introduction – Fall Semester 2024 Cell-cell junctions and cell migration at a glance Coordinator: Britta Engelhardt: [email protected] Introduction: Cell-cell junctions and cell migration at a glance > Part I General overview of the topic > Part II Introduction into the structure of the lecture 2 Definition of cell migration https://ki-galleries.mit.edu/2012/turner-2 Translocation of the cell from one location to another 3 Reasons to migrate: Unicellular organisms Food Mating Burnette et al. (2014) J. Cell Biol. 205:83-96 4 Reasons to migrate: Multicellular organisms > Gastrulation > Morphogenesis during embryonic development — Neural crest cells migrate long distances: from neural tube throughout the entire embryo — Coordinated movement of entire epithelial sheets > Organogenesis: Angiogenesis > Homeostasis: Gut and Immunosurveillance > Tissue repair, Inflammation > Disease — Tumor metastasis — Chronic inflammation 5 Move! How? 6 Migration in 2D Movement of cells on a solid substratum Move - how? Dictyostelium discoidum = amoeba 7 Prerequisite for migration Polarization 8 Microtubules determine and are essential for positioning the leading edge of a migrating cell Microtobule organizing center - MTOC 9 Cell Migration - 2D Movement of cells across a solid substratum Dictyostelium discoidum Pseudopods are often extended above the substratum Moving cell has limited contact with substratum 10 Movement of mutated Dictyostelium Pseudopod formation is not suppressed in the back Lack of polarization and thus no directed motility! 11 The migration cycle on 2D surfaces actin polymerisation-dependent processes cell-substrate adhesive structures myosin II dependent events (1) extension of the leading edge and formation of immature cell-substrate adhesions (2) maturation of cell-substrate adhesions (3) forward translocation of the cell body (4) disassembly of focal adhesions coupled to retraction of the rear edge Reig et al., Development, 2014: 141 12  Cell migration: Integration of signals from the front to the back of the cell 13 > Extensions of protrusions — actin cytoskeleton and regulators of actin dynamics regulate plasma membrane protrusions by de novo actin polymerisation into the direction of the movement > Adhesion — adhesion to the extracellular matrix — actin cytoskeleton is connected to the substratum via transmembrane adhesion molecules in the cell membrane – over 150 molecules identified in adhesions > Regulation and integration — local, transient signals retain polarity of the cell and orchestrate actin polymerization, adhesion, actomyosin bundling and contraction and microtubule dynamics. — Central role for signaling scaffolds that localize and activate kinases and phosphatases > Cell Body Translocation, Traction and Retraction at the rear end — Bulk of cytoplasma is pushed forward by actomyosin contraction — Disassembly of adhesions at the rear of a cell  Motility in 2D Ridley et al. 2003; Science 302: 1704 14 Players in Cell Polarization and Migration > Microtubules - essential for positioning of the leading edge - delivery of Golgi-derived vesicles to the leading edge > Actinomyosin > Rho family small GTPases: RhoA, Rac, Cdc42 - cdc42, a master regulator of cell polarity - localized Rac activation: initiating and maintaining protrusions > Rho family small GTPases: RhoA, Rac, Cdc42 > the gradient amplifiers > Integrins – mainly b1-integrins - activated integrins preferentially localize to the leading edge - stabilization of protrusions > Vesicles 15 Front-back polarity in a migrating cell: role of small GTPases Reig et al., Development, 2014: 141 regulate actin dynamics, adhesion organisation and protrusion formation 16 Small guanosine triphosphate (GTP) - binding proteins (GTPases): Controlling front-to-back polarity of migrating cells GAP GTPase activating protein inactivates Rho proteins GDI GDP dissociation inhibitor GEF guanine nucleotide exchange factor activates Rho proteins Bento et al. J Cell Sci 2013  Adhesions are mediated by integrins Avraamides et al, Nature Rev Cancer, 2008 18 Cells extend different plasma membrane protrusions at the leading edge > Lamellipodia and Filopodia — actin polymerization directly pushes the plasma membrane forward > Lobopodia – 3D only — actin containing rounded or cylindrical extensions protrude between extracellular matrix fibers > Invadopodia — actin polymerization and release of matrix degrading metalloproteases clear the path for the cell > Membrane blebs — plasma membrane is driven forward by cortical actomyosin contraction and reversible detachment of the plasma membrane from the cortical actin cytoskeleton 19 The Protrusive Machinery major difference is in actin organization and dynamics Lamellipodia (epithelial cells like keratocytes, fibroblasts) - flat- 2-dimensional - broad sheet of membrane and polymerized actin filaments flowing forward at the front of a cell in planar substrate — actin filaments form a branching “dendritic“ network involving Arp2/3 and WASP/WAVE - assembly by branched nucleation, Rac activation Filopodia (sprouting axons, dendritic cells) - early form = spikes – 1 dimensional — Core of actin filaments form long parallel bundles - assembly via elongation regulated by Cdc42 activation — Might serve as sensors, exploring local environment — (cave: not essential for chemotaxis) Invadopodia (neutrophils) actin rich matrix degrading protrusions Membrane blebbing drives directional migration of cells during embryonic development Lobopodia (leukocytes, amoeba) elongation of blebs, 3-dimensional, do not align with extracellular matrix 20 Fish keratocyte with lamellipodium Small et al, 1995 JCB 129 21 Growth cone of a sensory neuron with filopodia Photo by Ken Balazovich http://www.uni-leipzig.de/~pwm/web/?section=introduction&page=neurons 22 a | Podosomes and invadopodia are actin-rich structures formed on the ventral membrane of the cell. Podosomes and invadopodia can form as rosettes (left box) or individual puncta (right box), both of which are structures that protrude into the extracellular matrix (ECM). b | The image shows vascular smooth muscle cells (left) that were stimulated with platelet-derived growth factor as an example of a cell containing podosomes, and squamous carcinoma cells (right) as an example of a cell containing invadopodia. For both cell types, the image is shown both B&W and in colour. Classically, the presence of podosomes and invadopodia is confirmed by observing filamentous actin (F-actin) puncta colocalizing with sites of matrix degradation. The cells on top of the fluorescently conjugated matrix (gelatin labelled with fluorescein isothiocyanate (FITC); shown in green) are stained for F-actin using fluorescently conjugated phalloidin (white dots in B&W images and red overlay in colour images). The cells are then examined for colocalization between F-actin puncta (red dots) and matrix degradation (black regions); selected regions of colocalization are indicated by white arrows. The cell nuclei are shown in blue. Images courtesy of M. Quintavalle and B. Diaz, Sanford-Burnham Medical Research Institute, California, USA.  Membrane blebs A: Melanoma cell making membrane blebs. As a result, they crawl poorly and tend not to metastasize. B: Melanoma cells making lamellipodia are highly metastatic. From C. Cunningham et al., J. Cell Biol. 136:845–857, 1997 24  The migration cycle – 2D versus 3D Reig et al., Development, 2014: 141 25  Cell migration in 3D - within extracellular matrix in the presence of soluble guidance cues Mesenchymal migration Ameboid migration Renkawitz and Sixt, EMBO Reports, 2010 26 Cell migration in 3D - within extracellular matrix 27  Live cell imaging of cell migration in a 3D collagen network Mesenchymal migration Amoeboid migration 80 min tumor cell 10 min T cell Courtesy of Peter Friedl - Würzburg, Germany: Wolf, K., et al., Blood 160:267-277 (2003). 28 Adhesion and Traction in cells forming lamellipodia –mesenchymal migration > In cells forming lamellipodia integrins connecting ECM with actin cytoskeleton serve — as traction sites over which the cell moves — as mechanosensor – transmitting information about the physical state of the ECM into the cell and altering cytoskeletal dynamics. > Migration speed is dependent on — turnover-rate of adhesion and de-adhesion events — ability to detach — strength of cell attachment – density of adhesion receptors on the cells – affinity of the receptors for the adhesive ligands – High avidity increases attachment - reduces detachment - resulting in reduced migration rate > Adhesion strength is inversely correlated to speed of locomotion > High traction forces needed to detach integrins at the rear of the cell 29  Adhesion and Traction in lymphocytes – amoeboid migration > Adhesion to matrix is not necessary for locomotion because forward locomotion is a consequence of traction forces created in parallel to the cell membrane > Principle: force coupling receptors can generate traction without anchoring the cell – but cell needs to be in contact with the substrate- e.g. in 3D - scaffolds > In 3D scaffolds the cell has to squeeze between rigid structures enforcing contact with the substrate > 3D scaffolds therefore allow transmission of traction forces without the cell sticking to substrate > Traction forces required to move a non-adherent cell are low – cell has to move its mass and to overcome the viscous drag of the surrounding fluid 30  Live cell imaging of cell migration in a 3D collagen network Mesenchymal migration Amoeboid migration 80 min tumor cell 10 min T cell Courtesy of Peter Friedl - Würzburg, Germany: Wolf, K., et al., Blood 160:267-277 (2003). 31 Collective migration: complex cell-cell- and cell-ECM interactions Courtesy of P.Friedl 32  Multicellular migration of primary rhabdomyosarcoma explant in 3D collagen matrix 200 µm after 5 days of culture in 3-D collagen Friedl, P., et al., Cancer Res. 55:4557-4560 (1995) 33 Collective migration: complex cell-cell- and cell-ECM interactions Courtesy of P.Friedl 34 Cell contacts and cell motility Cell-to-Cell Adhesion and Communication 35  Junctions with different functions Laird, Biochem J. 2006, 394, 527 36 Introduction: Cell-cell junctions and cell migration at a glance Part II Introduction into the structure of the lecture 37 Dynamics of cellular contacts: Cell-cell contacts and cell motility Aim 38  Who is giving this lecture? Lecturers from the national PhD Program Cell Migration https://www.tki.unibe.ch/continuing_education/cellmigration/index_eng.html > Prof. Britta Engelhardt – Theodor Kocher Institute > Dr. Urban Deutsch – Theodor Kocher Institute > Prof. Ruth Lyck – Theodor Kocher Institute > Dr. Valentina Cecchinato, Institute for Research in Biomedicine, Bellinzona > Dr. Julia Gutjahr, Institute for Cell Biology and Immunology Thurgau, Kreuzlingen > Prof. Daniel Legler, Institute for Cell Biology and Immunology Thurgau, Kreuzlingen > Prof. Christoph Scheiermann, Department Pathology and Immunology, University of Geneva 39  Who is giving this lecture? Lecturers from the national PhD Program Cell Migration https://www.tki.unibe.ch/continuing_education/cellmigration/index_eng.html and Dr. Jérémie Rossy, Institute for Cell Biology and Immunology Thurgau, Kreuzlingen PD Dr. Martin Degen, Clinic for Orthodontics, University of Bern Dr. Paolo Armando Gagliardi, Institute for Cell Biology, University of Bern Prof. Olivier Pertz, Institute for Cell Biology, University of Bern Dr. Mykhailo Vladymyrov, Data Science Lab, University of Bern 40  Structure of the lecture – Part 1 Building knowledge on structures and function 1: Cell-cell junctions and cell migration at a glance – an introduction Introduction 3: Cytoskeleton: 4: Molecular Basic 2: Cell adhesion to defining cell composition and extracellular matrix structure and function of knowledge membrane cellular junctions organisation 41 Lecture 2: Cell adhesion to extracellular matrix Martin Degen 42 Lecture 3: Cytoskeleton - defining cell structure and membrane organisation Jérémy Rossy 43  Lecture 4: Molecular composition and function of cellular junctions Britta Engelhardt Green et al., 2019; https://doi.org/10.12688/f1000research.20942.1  Structure of the lecture – Part 1 Building knowledge on structures and function 1: Cell-cell junctions and cell migration at a glance – an introduction Introduction 3: Cytoskeleton: 4: Molecular Basic 2: Cell adhesion to defining cell composition and extracellular matrix structure and function of knowledge membrane cellular junctions organisation Function 7: From cells to 8a:Cell migration 5: Cell Motility I: tissues: role of in response to 8b:Chemokine ⚭ Polarization, 6: Cell motility II: the cytoskeleton chemokines and signaling protrusions, adhesive interactions and ⚭ less expected contributions to in development and homeostasis synergy-inducing molecules in pathways guiding cell retraction translate cell migration of epithelial physiology and migration into cell migration layers pathology 45 Lecture 5: Cell Motility I - Polarization, protrusions, adhesive interactions and retraction translate into cell migration Ruth Lyck 46 Lecture 5: Cell Motility I - Polarization, protrusions, adhesive interactions and retraction translate into cell migration Ruth Lyck 47 Lecture 6: Cell Motility II - Less expected contributions to cell migration Jérémy Rossy 48 Lecture 7: From cells to tissues: role of the cytoskeleton in development and homeostasis of epithelial layers Paolo Gagliardi 49 Lecture 8a: Cell migration in response to chemokines and synergy-inducing molecules in physiology and pathology Valentina Cecchinato 50 Lecture 8a: Cell migration in response to chemokines and synergy-inducing molecules in physiology and pathology Valentina Cecchinato 51 Lecture 8b: Chemokine signaling pathways guiding cell migration 52  End of Part 1: You have build knowledge on structures and function 1: Cell-cell junctions and cell migration at a glance – an introduction Introduction 3: Cytoskeleton: 4: Molecular Basic 2: Cell adhesion to defining cell composition and extracellular matrix structure and function of knowledge membrane cellular junctions organisation Function 7: From cells to 8a:Cell migration 5: Cell Motility I: tissues: role of in response to 8b:Chemokine ⚭ Polarization, 6: Cell motility II: the cytoskeleton chemokines and signaling protrusions, adhesive interactions and ⚭ less expected contributions to in development and homeostasis synergy-inducing molecules in pathways guiding cell retraction translate cell migration of epithelial physiology and migration into cell migration layers pathology 53 Structure of the lecture – Part 2 Deepening your knowledge with specific examples 10: Spatio-temporal 11: Guidance Rho GTPase molecules direct signaling to the cell movement cytoskeleton 12a: Cell migration in the 9: Immune cell development of migration and Select Topics the hematopoietic activation Knowledge system deepening 13a: The multi- 12b: Circadian step process of regulation of cell cancer metastasis migration 54 Lecture 9: Immune cell migration and activation Jérémy Rossy 55 Lecture 10: Spatio-temporal Rho GTPase signaling to the cytoskeleton 56 Lecture 11: Guidance molecules direct cell movement Urban Deutsch How do migrating cells find their targets? Migrating cells often follow chemotactic gradients of soluble molecules or move towards localized morphogens, growth factors or chemokine deposits Guidance molecules however direct cells along defined tracks Most guidance molecules execute these functions by direct interaction of transmembrane or membrane-bound proteins Determination of the exact migration route by a combination of attraction and repulsion Biological processes depending on guidance molecules We will discuss the families of guidance molecules involved Lecture 12a: Cell migration in the development of the hematopoietic system Julia Gutjahr 58 Lecture 12b: Circadian regulation of cell migration Christoph Scheiermann 59  Lecture 12b: Circadian regulation of cell migration Christoph Scheiermann 60 Lecture 13a: The multi-step process of cancer metastasis formation Ruth Lyck 61 End of Part 2: Deepened your knowledge with specific examples 10: Spatio-temporal 11: Guidance Rho GTPase molecules direct signaling to the cell movement cytoskeleton 12a: Cell migration in the 9: Immune cell development of migration and Select Topics the hematopoietic activation Knowledge system deepening 13a: The multi- 12b: Circadian step process of regulation of cell cancer metastasis migration 62 Structure of the lecture – Part 3 Experimental methods to study cell migration 14a: In vitro 14b: In vivo ⚭ 13b: Computational analysis of cell techniques to study techniques to study migration cell migration cell migration 63 Lecture 13b: Computational analysis of cell migration Mykhailo Vladymyrov 64 Lecture 14a: in vitro techniques to study cell migration Ruth Lyck 65 Lecture 14b: in vivo techniques to study cell migration Britta Engelhardt Vladymyrov et al. – Front Physics (Medical Physics and Imaging), 2020 https://doi.org/10.3389/fphy.2019.00222 Dynamics of cellular contacts: Cell-cell contacts and cell motility – Schedule 2024 Blue lectures will be in the Zoom classroom !! Aim 67