ERC Advanced Grant 741350

What Holds Cancer Cells Back?

 

in context of H2020

 

Since decades the bulk of cancer research focusses on the genetic and molecular level. To complement this knowledge, I will focus on the collective behaviour of cancer cells in cell clusters and in the extracellular matrix (ECM). Conventional cancer research tackles issues like genetic changes, signalling pathways or intracellular mechanisms. In contrast, I want to answer the fundamental, mechanistic question: When is a cancer cell jammed or when can it overcome the yield stress to actively „flow“ in a dense microenvironment (ME)? Within the concept of Physics of Cancer, I have brought forward the idea that changes in a cancer cell’s material properties determine its metastatic potential. To build upon this I propose the next breakthrough by determining a predictive phase diagram for unjamming transitions of cancer cells. Cancer cell jamming is quantified by cell speed as a measure of the motile forces and by cellular shape to account for the interplay between cell contractility and adhesion. Our self-propelled Voronoi model (SPV) will conclusively explain whether a cell is jammed by its neighbours or the ECM, thereby overcoming the limitations of existing theories which only apply to specific environments. Building on my leadership in cell biomechanics and the exclusive clinical access to two types of carcinomas (mamma, cervix), I will introduce highly innovative bionic modulators of intracellular mechanics and develop live cancer cell tracking in biopsies as a ground-breaking alternative to vital imaging. While these approaches are perfect to prove that unjamming transitions are key to tumour progression, I will investigate to what extent fluid, i.e. unjammed, tissue behaviour can be detected by magnetic resonance imaging elastography (MRE) as an individual predictive marker for metastasis. Moreover, the results may guide surgeons when considering the local spreading of cancer and thus greatly empower surgery in tumour therapies.

 

Events

Physics of Cancer

The aim of this symposium is by bringing together exceptional researchers in the areas of quantitative cell biology, physical mechanisms of pathology, cancer biology, molecular design, diagnostic systems and beyond to create a forum for the exchange of new ideas and formulation of new solutions.

2019

2018

2017


Publications

 

J Schnauß, BUS Schmidt, CB Brazel, S Dogan, W. Losert, U. Anderegg, JA Käs:

Influence of Hyaluronic Acid Binding on the Actin Cortex measured by Optical Forces Abstract:Melanoma cells are often surrounded by hyaluronic acid (HA) rich environments, which are considered to promote tumor progression and metastasis. Induced effects in compound materials consisting of cells embedded in an extracellular matrix (ECM) have been studied, however, alterations of the single cells have never been addressed. Here, we explicitly addressed single cell properties and measured HA-induced biomechanical changes via deformations induced solely by optical forces. With the optical stretcher setup, cells were deformed after culturing them either in the presence or absence of HA revealing the crucial interplay of HA with the CD44 receptor. To assess the role of CD44 in transducing effects of HA, we compared a CD44 expressing variant of the melanoma cell line RPM-MC to its natural CD44-negative counterpart. Our measurements revealed a significant stiffness change, which we attribute to changes of the actin cytoskeleton.

Journal of Biophotonics, e201960215 (2020),   DOI: 10.1002/jbio.20190215  

 

KJ Streitberger, L. Lilaj, F Schrank, J Braun, KT Hoffmann, M Reiss-Zimmermann, JA Käs, I Sack:

How tissue fluidity influences brain tumor progression Abstract:Mechanical properties of biological tissues and, above all, their solid or fluid behavior influence the spread of malignant tumors. While it is known that solid tumors tend to have higher mechanical rigidity, allowing them to aggressively invade and spread in solid surrounding healthy tissue, it is unknown how softer tumors can grow within a more rigid environment such as the brain. Here, we use in vivo magnetic resonance elastography (MRE) to elucidate the role of anomalous fluidity for the invasive growth of soft brain tumors, showing that aggressive glioblastomas (GBMs) have higher water content while behaving like solids. Conversely, our data show that benign meningiomas (MENs), which contain less water than brain tissue, are characterized by fluid-like behavior. The fact that the 2 tumor entities do not differ in their soft properties suggests that fluidity plays an important role for a tumor's aggressiveness and infiltrative potential. Using tissue-mimicking phantoms, we show that the anomalous fluidity of neurotumors physically enables GBMs to penetrate surrounding tissue, a phenomenon similar to Saffman-Taylor viscous-fingering instabilities, which occur at moving interfaces between fluids of different viscosity. Thus, targeting tissue fluidity of malignant tumors might open horizons for the diagnosis and treatment of cancer.

PNAS January 7, 117 (1) 128 - 134 (2020),   DOI: 10.1073/pnas.1913511116  

 

P van Liedekerke, J Neitsch, T Johann, E Warmt, IG Valverde, S Grosser, S Hoehme, JA Käs, D Drasdo:

Quantifying the mechanics and growth of cells and tissues in 3D using high resolution computational models Abstract:Mathematical models are increasingly designed to guide experiments in biology, biotechnology, as well as to assist in medical decision making. They are in particular important to understand emergent collective cell behavior. For this purpose, the models, despite still abstractions of reality, need to be quantitative in all aspects relevant for the question of interest. The focus in this paper is to study the regeneration of liver after drug-induced depletion of hepatocytes, in which surviving dividing and migrating hepatocytes must squeeze through a blood vessel network to fill the emerged lesions. Here, the cells' response to mechanical stress might significantly impact on the regeneration process. We present a 3D high-resolution cell-based model integrating information from measurements in order to obtain a refined quantitative understanding of the cell-biomechanical impact on the closure of drug-induced lesions in liver. Our model represents each cell individually, constructed as a physically scalable network of viscoelastic elements, capable of mimicking realistic cell deformation and supplying information at subcellular scales. The cells have the capability to migrate, grow and divide, and infer the nature of their mechanical elements and their parameters from comparisons with optical stretcher experiments. Due to triangulation of the cell surface, interactions of cells with arbitrarily shaped (triangulated) structures such as blood vessels can be captured naturally. Comparing our simulations with those of so-called center-based models, in which cells have a rigid shape and forces are exerted between cell centers, we find that the migration forces a cell needs to exert on its environment to close a tissue lesion, is much smaller than predicted by center-based models. This effect is expected to be even more present in chronic liver disease, where tissue stiffens and excess collagen narrows pores for cells to squeeze through.

Biomechanics and Modelling in Mechanobiology, 19, 189 - 220 (2020),   DOI: 10.1007/s10237-019-01204-7  

 

P M Shahryari, H Tzschätzsch, J Guo, SR Marticorena Garcia, G Böning, U Fehrenbach, L Stencel, P Asbach, B Hamm, JA Käs, J Braun, T Denecke, I Sack:

Tomoelastography Distinguishes Noninvasively between Benign and Malignant Liver Lesions Abstract:Patients with increased liver stiffness have a higher risk of developing cancer, however, the role of fluid-solid tissue interactions and their contribution to liver tumor malignancy remains elusive. Tomoelastography is a novel imaging method for mapping quantitatively the solid-fluid tissue properties of soft tissues in vivo. It provides high resolution and thus has clear clinical applications. In this work we used tomoelastography in 77 participants, with a total of 141 focal liver lesions of different etiologies, to investigate the contributions of tissue stiffness and fluidity to the malignancy of liver tumors. Shear-wave speed (c) as surrogate for tissue stiffness and phase-angle (f) of the complex shear modulus reflecting tissue fluidity were abnormally high in malignant tumors and allowed them to be distinguished from nontumorous liver tissue with high accuracy [c: AUC = 0.88 with 95% confidence interval (CI) = 0.83-0.94; f: AUC = 0.95, 95% CI = 0.92-0.98]. Benign focal nodular hyperplasia and hepatocellular adenoma could be distinguished from malignant lesions on the basis of tumor stiffness (AUC = 0.85, 95% CI = 0.72-0.98; sensitivity = 94%, 95% CI = 89-100; and specificity = 85%, 95% CI = 62-100), tumor fluidity (AUC = 0.86, 95% CI = 0.77-0.96; sensitivity = 83%, 95% CI = 72-93; and specificity = 92%, 95% CI = 77-100) and liver stiffness (AUC = 0.84, 95% CI = 0.74-0.94; sensitivity = 72%, 95% CI = 59-83; and specificity = 88%, 95% CI = 69-100), but not on the basis of liver fluidity. Together, hepatic malignancies are characterized by stiff, yet fluid tissue properties, whereas surrounding nontumorous tissue is dominated by solid properties. Tomoelastography can inform noninvasively on the malignancy of suspicious liver lesions by differentiating between benign and malignant lesions with high sensitivity based on stiffness and with high specificity based on fluidity.

Cancer Research vol. 79 iss. 22 (2019),   DOI: 10.1158/0008-5472.CAN-19-2150  

 

H Kubitschke, B Wolf, E Morawetz, L-C Horn, B Aktas, U Behn, M Höckel, JA Käs:

Roadmap to Local Tumour Growth: Insights from Cervical Cancer Abstract:Wide tumour excision is currently the standard approach to surgical treatment of solid cancers including carcinomas of the lower genital tract. This strategy is based on the premise that tumours exhibit isotropic growth potential. We reviewed and analysed local tumour spreading patterns in 518 patients with cancer of the uterine cervix who underwent surgical tumour resection. Based on data obtained from pathological examination of the surgical specimen, we applied computational modelling techniques to simulate local tumour spread in order to identify parameters influencing preferred infiltration patterns and used area-proportional Euler diagrams to detect and confirm ordered patterns of tumour spread. Some anatomical structures, e.g. tissues of the urinary bladder, were significantly more likely to be infiltrated than other structures, e.g. the ureter and the rectum. Computational models assuming isotropic growth could not explain these infiltration patterns. Introducing ontogenetic distance of a tissue relative to the uterine cervix as a parameter led to accurate predictions of the clinically observed infiltration likelihoods. The clinical data indicates that successive infiltration likelihoods of ontogenetically distant tissues are nearly perfect subsets of ontogenetically closer tissues. The prevailing assumption of isotropic tumour extension has significant shortcomings in the case of cervical cancer. Rather, cervical cancer spread seems to follow ontogenetically defined trajectories.

Scientific Reports 9, Article number 12768 (2019),   DOI: 10.1038/s41598-019-49182-1  

 

C Ficorella, R Martinez Vàzques, P Heine, E Lepera, J Cao, E Warmt, R Osellame, JA Käs:

Normal epithelial and triple-negative breast cancer cells show the same invasion potential in rigid spatial confinement Abstract:The extra-cellular microenvironment has a fundamental role in tumor growth and progression, strongly affecting the migration strategies adopted by single cancer cells during metastatic invasion. In this study, we use a novel microfluidic device to investigate the ability of mesenchymal and epithelial breast tumor cells to fluidize and migrate through narrowing microstructures upon chemoattractant stimulation. We compare the migration behavior of two mesenchymal breast cancer cell lines and one epithelial cell line, and find that the epithelial cells are able to migrate through the narrowest microconstrictions as the more invasive mesenchymal cells. In addition, we demonstrate that migration of epithelial cells through a highly compressive environment can occur in absence of a chemoattractive stimulus, thus evidencing that they are just as prone to react to mechanical cues as invasive cells.

New Journal of Physics, 21 (2019),   DOI: 10.1088/1367-2630/ab3572  

 

T Golde, M Glaser, C Tutmarc, I Elbalasy, C Huster, G Busteros, DM Smith, H Herrmann, JA Käs, J Schnauß:

The role of stickiness in the rheology of semiflexible polymers Abstract:Semiflexible polymers form central structures in biological material. Modelling approaches usually neglect influences of polymer-specific molecular features aiming to describe semiflexible polymers universally. Here, we investigate the influence of molecular details on networks assembled from filamentous actin, intermediate filaments, and synthetic DNA nanotubes. In contrast to prevalent theoretical assumptions, we find that bulk properties are affected by various inter-filament interactions. We present evidence that these interactions can be merged into a single parameter in the frame of the glassy wormlike chain model. The interpretation of this parameter as a polymer specific stickiness is consistent with observations from macro-rheological measurements and reptation behaviour. Our findings demonstrate that stickiness should generally not be ignored in semiflexible polymer models.

Soft Matter, 15, 4865-4872 (2019),   DOI: 10.1039/C9SM00433E 

 

F Sauer, L Oswald, A Ariza de Schellenberger, H Tzschätzsch, F Schrank, T Fischer, J Braun, CT Mierke, R Valiullin, I Sack, JA Käs:

Collagen networks determine viscoelastic properties of connective tissues yet do not hinder diffusion of the aqueous solvent Abstract:Collagen accounts for the major extracellular matrix (ECM) component in many tissues and provides mechanical support for cells. Magnetic Resonance (MR) Imaging, MR based diffusion measurements and MR Elastography (MRE) are considered sensitive to the microstructure of tissues including collagen networks of the ECM. However, little is known whether water diffusion interacts with viscoelastic properties of tissues. This study combines highfield MR based diffusion measurements, novel compact tabletop MRE and confocal microscopy in collagen networks of different cross-linking states (untreated collagen gels versus additional treatment with glutaraldehyde). The consistency of bulk rheology and MRE within a wide dynamic range is demonstrated in heparin gels, a viscoelastic standard for MRE. Additional crosslinking of collagen led to an 8-fold increased storage modulus, a 4-fold increased loss modulus and a significantly decreased power law exponent, describing multi-relaxational behavior, corresponding to a pronounced transition from viscous-soft to elastic-rigid properties. Collagen network changes were not detectable by MR based diffusion measurements and microscopy which are sensitive to the micrometer scale. The MRE-measured shear modulus is sensitive to collagen fiber interactions which take place on the intrafiber level such as fiber stiffness. The insensitivity of MR based diffusion measurements to collagen hydrogels of different cross-linking states alludes that congeneric collagen structures in connective tissues do not hinder extracellular diffusive water transport. Furthermore, the glutaraldehyde induced rigorous changes in viscoelastic properties indicate that intrafibrillar dissipation is the dominant mode of viscous dissipation in collagen-dominated connective tissue.

Soft Matter, 15, 3055-3064 (2019),   DOI: 10.1039/C8SM02264J 

 

CT Mierke, F Sauer, S Grosser, S Puder, T Fischer, JA Käs:

The two faces of enhanced stroma: Stroma acts as a tumor promoter and a steric obstacle Abstract: In addition to genetic, morphological and biochemical alterations in cells, a key feature of the malignant progression of cancer is the stroma, including cancer cell motility as well as the emergence of metastases. Our current knowledge with regard to the biophysically driven experimental approaches of cancer progression indicates that mechanical aberrations are major contributors to the malignant progression of cancer. In particular, the mechanical probing of the stroma is of great interest. However, the impact of the tumor stroma on cellular motility, and hence the metastatic cascade leading to the malignant progression of cancer, is controversial as there are two different and opposing effects within the stroma. On the one hand, the stroma can promote and enhance the proliferation, survival and migration of cancer cells through mechanotransduction processes evoked by fiber alignment as a result of increased stroma rigidity. This enables all types of cancer to overcome restrictive biological capabilities. On the other hand, as a result of its structural constraints, the stroma acts as a steric obstacle for cancer cell motility in dense three-dimensional extracellular matrices, when the pore size is smaller than the cell´s nucleus. The mechanical properties of the stroma, such as the tissue matrix stiffness and the entire architectural network of the stroma, are the major players in providing the optimal environment for cancer cell migration. Thus, biophysical methods determining the mechanical properties of the stroma, such as magnetic resonance elastography, are critical for the diagnosis and prediction of early cancer stages. Fibrogenesis and cancer are tightly connected, as there is an elevated risk of cancer on cystic fibrosis or, subsequently, cirrhosis. This also applies to the subsequent metastatic process.

NMR Biomed., vol. 10, e3831 (2018),  DOI: 10.1002/nbm.3831

 

T Golde, C Huster, M Glaser, T Händler, H Herrmann, JA Käs, J Schnauß:

Glassy dynamics in composite biopolymer networks Abstract: The cytoskeleton is a highly interconnected meshwork of strongly coupled subsystems providing mechanical stability as well as dynamic functions to cells. To elucidate the underlying biophysical principles, it is central to investigate not only one distinct functional subsystem but rather their interplay as composite biopolymeric structures. Two of the key cytoskeletal elements are actin and vimentin filaments. Here, we show that composite networks reconstituted from actin and vimentin can be described by a superposition of two non-interacting scaffolds. Arising effects are demonstrated in a scale-spanning frame connecting single filament dynamics to macro-rheological network properties. The acquired results of the linear and non-linear bulk mechanics can be captured within an inelastic glassy wormlike chain model. In contrast to previous studies, we find no emergent effects in these composite networks. Thus, our study paves the way to predict the mechanics of the cytoskeleton based on the properties of its single structural components.

Soft Matter vol. 14, 7970-7978 (2018),  DOI: 10.1039/C8SM01061G

 

F Meinhövel, R Stange, J Schnauß, M Sauer, JA Käs, TW Remmerbach:

Changing cell mechanics - a precondition for malignant transformation of oral squamous carcinoma cells Abstract: Oral squamous cell carcinomas (OSCC) are the sixth most common cancer and the diagnosis is often belated for a curative treatment. The reliable and early differentiation between healthy and diseased cells is the main aim of this study in order to improve the quality of the treatment and to understand tumour pathogenesis. Here, the optical stretcher is used to analyse mechanical properties of cells and their potential to serve as a marker for malignancy. Stretching experiments revealed for the first time that cells of primary OSCCs were deformed by 2.9% rendering them softer than cells of healthy mucosa which were deformed only by 1.9%. Furthermore, the relaxation behaviour of the cells revealed that these malignant cells exhibit a faster contraction than their benign counterparts. This suggests that deformability as well as relaxation behaviour can be used as distinct parameters to evaluate emerging differences between these benign and malignant cells. Since many studies in cancer research are performed with cancer cell lines rather than primary cells, we have compared the deformability and relaxation of both types, showing that long time culturing leads to softening of cells. The higher degree of deformability and relaxation behaviour can enable cancer cells to traverse tissue emphasizing that changes in cell architecture may be a potential precondition for malignant transformation. Respecting the fact that even short culture times have an essential effect on the significance of the results, the use of primary cells for further research is recommended. The distinction between malignant and benign cells would enable an early confirmation of cancer diagnoses by testing cell samples of suspect oral lesions.

Convergent Science Physical Oncology, vol. 4, 034001(2018),  DOI: 10.1088/2057-1739/aac72d

 

J Lorenz, J Schnauß, M Glaser, M Sajfutdinow, C Schuldt, JA Käs, DM Smith:

Synthetic Transient Crosslinks Program the Mechanics of Soft, Biopolymer-Based Materials Abstract: Actin networks are adaptive materials enabling dynamic and static functions of living cells. A central element for tuning their underlying structural and mechanical properties is the ability to reversibly connect, i.e., transiently crosslink, filaments within the networks. Natural crosslinkers, however, vary across many parameters. Therefore, systematically studying the impact of their fundamental properties like size and binding strength is unfeasible since their structural parameters cannot be independently tuned. Herein, this problem is circumvented by employing a modular strategy to construct purely synthetic actin crosslinkers from DNA and peptides. These crosslinkers mimic both intuitive and noncanonical mechanical properties of their natural counterparts. By isolating binding affinity as the primary control parameter, effects on structural and dynamic behaviors of actin networks are characterized. A concentration-dependent triphasic behavior arises from both strong and weak crosslinkers due to emergent structural polymorphism. Beyond a certain threshold, strong binding leads to a nonmonotonic elastic pulse, which is a consequence of self-destruction of the mechanical structure of the underlying network. The modular design also facilitates an orthogonal regulatory mechanism based on enzymatic cleaving. This approach can be used to guide the rational design of further biomimetic components for programmable modulation of the properties of biomaterials and cells.

Advanced Materials 1706092 (2018),  DOI: 10.1002/adma.201706092

 

L Oswald, S Grosser, DM Smith, JA Käs:

Jamming transitions in cancer Abstract: The traditional picture of tissues, where they are treated as liquids defined by properties such as surface tension or viscosity has been redefined during the last few decades by the more fundamental question: Under which conditions do tissues display liquid-like or solid-like behaviour? As a result, basic concepts arising from the treatment of tissues as solid matter, such as cellular jamming and glassy tissues, have shifted into the current focus of biophysical research. Here, we review recent works examining the phase states of tissue with an emphasis on jamming transitions in cancer. When metastasis occurs, cells gain the ability to leave the primary tumour and infiltrate other parts of the body. Recent studies have shown that a linkage between an unjamming transition and tumour progression indeed exists, which could be of importance when designing surgery and treatment approaches for cancer patients.

Journal of Physics D: Applied Physics, Volume 50, 483001 (2017),  DOI: 10.1088/1361-6463/aa8e83

 

D Strehle, P Mollenkopf, M Glaser, T Golde, C Schuldt, JA Käs, J Schnauß:

Single Actin Bundle Rheology Abstract:Bundled actin structures play an essential role in the mechanical response of the actin cytoskeleton in eukaryotic cells. Although responsible for crucial cellular processes, they are rarely investigated in comparison to single filaments and isotropic networks. Presenting a highly anisotropic structure, the determination of the mechanical properties of individual bundles was previously achieved through passive approaches observing bending deformations induced by thermal fluctuations. We present a new method to determine the bending stiffness of individual bundles, by measuring the decay of an actively induced oscillation. This approach allows us to systematically test anisotropic, bundled structures. Our experiments revealed that thin, depletion force-induced bundles behave as semiflexible polymers and obey the theoretical predictions determined by the wormlike chain model. Thickening an individual bundle by merging it with other bundles enabled us to study effects that are solely based on the number of involved filaments. These thicker bundles showed a frequency-dependent bending stiffness, a behavior that is inconsistent with the predictions of the wormlike chain model. We attribute this effect to internal processes and give a possible explanation with regard to the wormlike bundle theory.

Molecules 22(10) 1804 (2017),   DOI: 10.3390/molecules22101804