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New technologies in cancer mechanobiology

20 - 21 June 2018 09:00 - 17:00

Satellite meeting organised by Dr Chris Bakal and Dr Julia Sero

The development of new technologies has long been a driving force in the study of mechanobiology. This workshop will bring together an interdisciplinary group of early to senior level researchers to discuss cutting-edge in vivo, in vitro, and in silico techniques and how these methods can be applied to cancer research, diagnostics, and therapeutics.

Recorded audio of the presentations will be available on this page after the meeting has taken place. The speaker biographies and abstracts are available below. 

Enquires: Contact the Scientific Programmes team.

Organisers

  • Dr Chris Bakal, Institute of Cancer Research, UK

    Chris Bakal is a Reader of Cell Form at the Institute of Cancer Research. He earned his BSc in Biochemistry from the University of British Columbia, and a PhD in Medical Biophysics from the University of Toronto. Chris’ postdoctoral work was performed in the Department of Genetics at Harvard Medical School, and the Computer Science and Artificial Intelligence Laboratory (CSAIL) at the Massachusetts Institute of Technology (MIT). His laboratory’s research is aimed at understanding how changes in cell shape drive tumorigenesis and metastasis. Towards these goals the Bakal laboratory uses quantitative single cell imaging technology, bioengineering, and computational methods. Dr Bakal’s contributions to the field of cancer cell biology include: the finding that cell shape can be used to infer the state of underlying signalling and transcriptional networks; that cell morphogenesis follows principles of dynamical systems; and that cell shape regulates the activity of key signalling proteins. These contributions have impacted our understanding of the role of cell shape changes in disease. 

    Chris was awarded the 2015 Cancer Research UK Future Leaders Prize. In 2014, he was awarded the Council for Systems Biology Merrimack Pharmaceuticals Prize, and in 2013 the British Association for Cancer Research Frank Rose Award. In 2007, Chris was awarded the Dorsett L. Spurgeon prize as one of the most promising postdoctoral fellows or junior faculty members at Harvard Medical School. 

    Outside of science Chris is a competitive track cyclist, and a former world-ranked downhill ski racer. 

  • Dr Julia Sero, Institute of Cancer Research, UK

    Dr Julia Sero studied cell and molecular biology at Penn State University and the University of Glasgow, and worked as an intern for many summers in microscopy and tissue engineering labs at Carnegie Mellon University. She then worked as a research assistant at MIT studying the genetics of cancer and development in mice with Jackie Lees. Dr Sero did her PhD with Donald Ingber at Harvard Medical School, using micropatterning to study cell adhesion and directional migration. In 2010, Julia joined Chris Bakal at the Institute of Cancer Research to investigate the dynamics of mechanosensitive transcriptional regulators as a function of cell shape. She is currently working with Molly Stevens's group at Imperial College to investigate cell movement and signalling in the context of bioengineered materials.

Schedule

09:05 - 09:15 Mechanobiology-based technology for rapid cancer diagnosis and prognosis

Metastasis requires cells to dynamically adapt to changing microenvironments and apply forces. Daphne Weihs shows that subpopulations of metastatic cells rapidly (<2 hours) and forcefully indent elastic, physiological stiffness, synthetic, impenetrable gels to depths of 1–20 µm, whereas benign/normal cells do not indent. The indenting subpopulation is highly migratory and invasive (Boyden chamber) and includes chemotherapy-resistant and cancer stem cells. Indentation capacity is governed by mechanobiology and is applicable to various solid cancers. Daphne will demonstrate with breast, pancreas, and skin cancers, rapid, same-day diagnosis and prognosis that matches the metastatic risk and clinical outcome in patients.

Professor Daphne Weihs, Technion-Israel Institute of Technology, Israel

09:15 - 09:30 Using mesoscopic models to understand the physical behaviours of cells and tissues

Living cells generate and transmit mechanical forces over diverse time-scales and length-scales to determine the dynamics of cell and tissue shape during both homeostatic and pathological processes. On the molecular scale, Mike Murrell’s group uses active gels as a framework to understand how mechanical stresses are transmitted within the cell cytoskeleton. On the scale of cells and tissues, they abstract these stresses to surface tensions in a liquid film and draw analogies between the dynamics of wetting to the shape dynamics of simple tissues. Together, they develop comprehensive descriptions for how cytoskeletal stresses translate to the physical behaviours of cells and tissues.

Dr Mike Murrell, Yale University, USA

09:30 - 09:45 Development of biomimetic stroma for 3D tumouroids

There has been a drive to develop 3D models to test mechanisms of cancer progression. Such models aim to recapitulate specific aspects of the native microenvironment of cancer tissue. Umber Cheema’s group has developed 3D models of solid cancers, termed tumouroids. Using tissue engineering techniques, they control the spatial positioning of a cancer mass and its surrounding stroma. The group are able to engineer specific components into each component. They have engineered hypoxic gradients within the 3D model as well as engineering an intact primitive vascular network in the stromal component. They have observed interaction of cancer cells with engineered vascular networks- and measured angiogenic remodelling of the networks. By incorporating cancer associated fibroblasts from patient samples Umber is further investigating how these cells enhance cancer invasion and the mechanisms by which this is done.

Dr Umber Cheema, University College London, UK

09:45 - 10:00 Mechanobiology in a microplate – opportunities for high throughput screening

Mechanobiology focuses on understanding how physical forces correlate with protein, cell and tissue dynamics and organization through mechano-transduction. Single molecule force measurements have revealed how force is used in biological systems, from individual proteins to complexes. These changes occur through alteration of biochemical properties, yet such properties cannot be readily measured alongside force manipulation experiments. To this end Chris Toseland has developed a novel mechanobiology assay format to encompass mechanical measurements with biochemical and cellular assays using a modified microplate system. He demonstrates his group’s approach using force-induced in vitro enzymatic activity and conformation changes, along with receptor activation of signalling pathways in live cells.

Dr Chris Toseland, University of Kent, UK

10:00 - 10:30 Discussion

Professor Daphne Weihs, Technion-Israel Institute of Technology, Israel

Dr Mike Murrell, Yale University, USA

Dr Umber Cheema, University College London, UK

Dr Chris Toseland, University of Kent, UK

10:30 - 11:00 Coffee break
00:00 - 00:00 Sparks, waves, well-plates and 3D tissue culture: high speed light sheet fluorescence microscopy in space and time

Light sheet fluorescence microscopy (LSFM) provides high speed low out-of-plane photobleaching and phototoxicity, but standard LSFM requires two microscope objective lenses orientated at 90° to one another to image the sample and this prevents imaging of conventional microscope slides and 96-well plates. Oblique plane microscopy (OPM) is an alternative approach that uses a single high numerical aperture microscope objective to provide both fluorescence excitation and detection, whilst maintaining the advantages of LSFM. Results of video-rate 3D imaging of calcium dynamics in cardiac myocytes and of cell migration in a 3D 96-well plate assay will be presented.

Dr Chris Dunsby, Imperial College London, UK

11:00 - 11:15 Toward systems biophotonics: imaging biology across high spatio-temporal dimensions and scales

The distribution and interactions of single molecules in three-dimensionally organised cellular networks are fundamental to the function of living systems. Today, we still lack a complete understanding of how local molecular mechanisms are integrated and dynamically mapped over larger scales on to functional activities. The challenges pose high demands for imaging technologies to provide molecular specificity, nanometre-scale resolution, ultrafast speed, and accessibility across larger volumes of tissues. In this presentation, Shu Jia will talk about his laboratory’s recently developed super-resolution and light-field microscopy, and functional imaging tools for high-throughput extraction of molecular information in cells and tissues with ultrahigh-spatiotemporal resolution and accessibility.

Dr Shu Jia, Stony Brook University, the State University of New York (SUNY), USA

11:30 - 11:45 Multiparametric in vivo microscopy of tumour-microenvironment interactions

Third harmonic generation microscopy (THGM) is a versatile label free imaging modality for intravital microscopy, revealing discontinuities of the refractive index in the specimen and suited to visualise microvesicles, nerves, basement membranes, collagen structures and muscle fibres. Here, it is shown how optimised TGHM can be combined with second harmonics generation and multiphoton fluorescence microscopy. It is demonstrated that tissue structures of the tumour microenvironment assist guided migration, but also limit tumour cell migration by induction of nuclear damage. Finally, THGM imaging depth is extended into previously hidden areas of the tumour, by application of high pulse energy excitation sources.

Dr Gert-Jan Bakker, Radboud Institute for Molecular Life Sciences, The Netherlands

11:45 - 12:15 Discussion

Dr Shu Jia, Stony Brook University, the State University of New York (SUNY), USA

Dr Chris Dunsby, Imperial College London, UK

Dr Gert-Jan Bakker, Radboud Institute for Molecular Life Sciences, The Netherlands

13:15 - 13:30 Mass spectrometry proteomics for mechanobiology

Proteins are the molecular machines that perform fundamental functional, structural and signalling roles in our cells and extracellular matrix. Protein regulation is therefore essential to the maintenance of tissue homeostasis, and the ability to measure the proteome can aid our understanding of ageing and disease processes. This presentation will examine how advances in label-free mass spectrometry proteomics can be applied to identify and quantify proteins in complex cell or tissue samples, and explore the relationship between proteins, mechano-signalling and mechanical properties.

Dr Joe Swift, Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, UK

13:30 - 13:45 Motility, mechanics and microscopes: local activation of TRPV4 channel at focal adhesions mediates cell mechanosensing and migration

The ability of cells to migrate toward areas of higher extracellular matrix (ECM) rigidity, durotaxis, plays a key role in various physiological processes ranging from immune responses to embryo development. This ability is also central to pathological conditions such as inflammatory diseases and cancer. To understand how cells sample ECM rigidity to migrate along ECM stiffness gradients, Sergey Plotnikov characterised calcium signalling at integrin-based focal adhesions. Sergey shows that focal adhesions are centres of transient calcium sparks via mechano-gated ion channel TRPV4. He demonstrates that these sparks are driven by cellular actomyosin contractility and dynamic traction forces exerted by focal adhesions on the ECM. Sergey found that calcium sparks are increased in cells plated on soft ECM allowing local ECM stiffness probing. He also shows that calcium sparks regulate focal adhesion turnover as well as directed migration towards stiffer ECM. He concludes that local activation of TRPV4 channels regulated by ECM stiffness promotes selective disassembly of the focal adhesions attached to softer ECM and thus provides a gradient of focal adhesion turnover across a migrating cell, which mediates durotaxis.

Dr Sergey Plotnikov, University of Toronto, Canada

13:45 - 14:00 Probing epithelial mechanics in 3D

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. Technologies to map 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, will be presented. Direct measurements of epithelial traction, tension, and luminal pressure in three-dimensional epithelia of controlled size and shape will also be presented.

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Professor Xavier Trepat, Institute for Bioengineering of Catalonia, Spain

14:00 - 14:30 Discussion

Dr Joe Swift, Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, UK

Dr Sergey Plotnikov, University of Toronto, Canada

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Professor Xavier Trepat, Institute for Bioengineering of Catalonia, Spain

14:30 - 15:00 Tea
15:00 - 15:15 Tracing actin polarity in cellular networks

The ability of a eukaryotic cell to resist deformation, to transport intracellular cargo and to change shape depends on the cytoskeleton, an interconnected network of filamentous polymers and regulatory proteins. In order to understand these processes, the directionality of actin filaments and their interactions with binding partners are of major importance. In this talk, Matthias Eibauer discusses how his team developed the means to detect actin directionality within unlabelled filamentous networks. This approach is based on cryo-electron tomography in conjunction with image processing. This method can be adopted to reveal the polarity and binders of actin filaments in cells and at physiological cellular processes.

Dr Matthias Eibauer, University of Zurich, Switzerland

15:15 - 15:30 Multiscale models to uncover the systems-mechanobiology of cancer

The field of mechanobiology uncovered a large number of ways how mechanical properties or forces may affect intracellular signalling pathways, eg through conformal deformations of proteins or the opening of ion gates. Such pathways, in turn, may also regulate the cytoskeleton or other mechanically active components of cells, leading to mechanochemical feedbacks. In this talk, mathematical modelling approaches are discussed that describe different aspects of mechanobiology. For instance, ODE models are employed to understand the pathway dynamics, PDE models are used to understand the effect of spatial localisation of signalling molecules, and stochastic models are used to describe the force-dependent dynamics of adhesion binding. Fabian Spill finally outlines approaches to combine different models, eg adhesion dynamics with cell and tissue forces, to gain a systems-level understanding of the mechanobiology of cancer.

Dr Fabian Spill, University of Birmingham, UK

15:30 - 15:45 Reverse engineering cell competition

Professor Guillaume Charras, University College London, UK

15:45 - 16:00 Discussion

Dr Matthias Eibauer, University of Zurich, Switzerland

Professor Guillaume Charras, University College London, UK

16:00 - 18:00 Breakout sessions I
09:30 - 09:45 Cell motility and extracellular basement membrane development in a model of breast development and breast cancer

Loss of the basement membrane is a hallmark of invasive cancer. Claire Robertson’s group studies how basement membrane develops by culturing nonmalignant breast epithelial cells (BEC), which develop into polarized, growth arrested structures to malignant breast cancer cells (BCC), which form disorganised masses under the same conditions. These two cell types show different movement patterns in the ECM gels: BEC rotate around their centre of mass in a process termed coherent angular motion, whereas BCC migrate randomly. Coherent angular motion appears to affect concentration of exogenous ECM by physically pulling ECM proteins into an organised shell at the cell surface as an early aspect of formation of a basement membrane-like ECM structure in mammary epithelial cells in culture, whereas this process is defective in malignant cells undergoing random motility.

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Dr Claire Robertson, Lawrence Berkeley National Lab, USA

09:45 - 10:00 CAFs: useful cellular models to investigate cancer mechanobiology

Cancer-associated fibroblasts (CAFs) are predominant non-malignant cells in tumours that actively remodel the extracellular matrix thereby affecting the physical properties of solid tumours. Fernando Calvo will show technologies to assess CAF function in cancer and how he has used them to unveil important aspects of cancer mechanobiology. In addition, he will discuss how he has used CAFs to decipher the molecular mechanisms governing mechanotransduction, and their impact in CAF behaviour and cancer progression.

Dr Fernando Calvo, Institute of Biomedicine & Biotechnology of Cantabria, Spain

10:00 - 10:15 YAP/TAZ in development, regeneration, and cancer

The YAP and TAZ transcriptional co-activators drive TEAD-dependent transcription to promote cell proliferation during normal development, tissue regeneration, and cancer. Barry Thompson will describe how YAP/TAZ are regulated by growth factor signalling, the Hippo pathway, mechanical forces and tissue damage in epithelia. He will further describe genetic loss-of-function and gain-of-function analysis of YAP/TAZ in both Drosophila and mice, and the implications of these findings for various types of human cancer.

Dr Barry Thompson, Francis Crick Institute, UK

10:15 - 10:30 Discussion
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Dr Claire Robertson, Lawrence Berkeley National Lab, USA

Dr Fernando Calvo, Institute of Biomedicine & Biotechnology of Cantabria, Spain

10:30 - 11:00 Coffee
11:00 - 11:15 Fast super-resolution imaging without artefacts

Conventional localisation microscopy relies on sparse activation of flurophores to allow accurate data fitting, meaning acquisition is slow and live cell experiments difficult or impossible. Several algorithms have been developed to cope with high emitter density. However, these produce significant image artefacts as the density is increased, which are easily mistaken for high resolution. By examining known biological structures it is shown that artefacts can be largely eliminated by pre-processing the image sequence with a succession of Haar wavelet kernels (HAWK), improving the resolving power and ensuring that the image reflects the structure of the sample.

Dr Susan Cox, King's College London, UK

11:15 - 11:30 Next-generation spatial RNA sequencing technologies

Tumour growth is regulated by cell autonomous as well as short- and long-range cell-cell interactions. The Lee group has been developing methods to label, track, and isolate single cells and their gene expression products in tumour tissues, including sequencing of somatic mutations, lineage reporters, and transcriptional markers. First, the talk will focus on a high-sensitivity in situ sequencing method for fluorescently labelling single cells and their applications in cancer medicine. Second, the talk will present paired-end DNA sequencing chemistry for 3D transcriptome reconstruction of developing tissues, concluding with discussing ways to study genotype-phenotype correlations in vivo.

Dr Je H Lee, Cold Spring Harbor Laboratory, USA

11:30 - 12:00 Discussion

Dr Susan Cox, King's College London, UK

Dr Je H Lee, Cold Spring Harbor Laboratory, USA