How cells sense tissue stiffness to guide cell migration is a fundamental question in development, fibrosis and cancer. Although durotaxis—cell migration towards increasing substrate stiffness—is well established, it remains unknown whether individual cells can migrate towards softer environments. Here, using microfabricated stiffness gradients, we describe the directed migration of U-251MG glioma cells towards less stiff regions. This ‘negative durotaxis’ does not coincide with changes in canonical mechanosensitive signalling or actomyosin contractility. Instead, as predicted by the motor–clutch-based model, migration occurs towards areas of ‘optimal stiffness’, where cells can generate maximal traction. In agreement with this model, negative durotaxis is selectively disrupted and even reversed by the partial inhibition of actomyosin contractility. Conversely, positive durotaxis can be switched to negative by lowering the optimal stiffness by the downregulation of talin—a key clutch component. Our results identify the molecular mechanism driving context-dependent positive or negative durotaxis, determined by a cell’s contractile and adhesive machinery.
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The data supporting the findings of this study are available within the Article and its Supplementary Information. Other raw data generated during this study are available from the corresponding authors on request. Source data are provided with this paper.
All code and scripts used in this study are available online (https://oddelab.umn.edu/ and via GitHub at https://github.com/cbcbcbcb123/Growth-Cone-Dynamics) and on request from the corresponding authors.
Ladoux, B., Mège, R.-M. & Trepat, X. Front–rear polarization by mechanical cues: from single cells to tissues. Trends Cell Biol. 26, 420–433 (2016).
Hamidi, H. & Ivaska, J. Every step of the way: integrins in cancer progression and metastasis. Nat. Rev. Cancer 18, 533–548 (2018).
Lo, C. M., Wang, H. B., Dembo, M. & Wang, Y. L. Cell movement is guided by the rigidity of the substrate. Biophys. J. 79, 144–152 (2000).
Isenberg, B. C., DiMilla, P. A., Walker, M., Kim, S. & Wong, J. Y. Vascular smooth muscle cell durotaxis depends on substrate stiffness gradient strength. Biophys. J. 97, 1313–1322 (2009).
Plotnikov, S. V., Pasapera, A. M., Sabass, B. & Waterman, C. M. Force fluctuations within focal adhesions mediate ECM-rigidity sensing to guide directed cell migration. Cell 151, 1513–1527 (2012).
Breckenridge, M. T., Desai, R. A., Yang, M. T., Fu, J. & Chen, C. S. Substrates with engineered step changes in rigidity induce traction force polarity and durotaxis. Cell. Mol. Bioeng. 7, 26–34 (2014).
Sunyer, R. et al. Collective cell durotaxis emerges from long-range intercellular force transmission. Science 353, 1157–1161 (2016).
DuChez, B. J., Doyle, A. D., Dimitriadis, E. K. & Yamada, K. M. Durotaxis by human cancer cells. Biophys. J. 116, 670–683 (2019).
Barriga, E. H., Franze, K., Charras, G. & Mayor, R. Tissue stiffening coordinates morphogenesis by triggering collective cell migration in vivo. Nature 554, 523–527 (2018).
Zhu, M. et al. Spatial mapping of tissue properties in vivo reveals a 3D stiffness gradient in the mouse limb bud. Proc. Natl Acad. Sci. USA 117, 4781–4791 (2020).
McKenzie, A. J. et al. The mechanical microenvironment regulates ovarian cancer cell morphology, migration, and spheroid disaggregation. Sci. Rep. 8, 7228 (2018).
Yeoman, B. et al. Adhesion strength and contractility enable metastatic cells to become adurotactic. Cell Rep. 34, 108816 (2021).
Hartman, C. D., Isenberg, B. C., Chua, S. G. & Wong, J. Y. Vascular smooth muscle cell durotaxis depends on extracellular matrix composition. Proc. Natl Acad. Sci. USA 113, 11190–11195 (2016).
Lachowski, D. et al. FAK controls the mechanical activation of YAP, a transcriptional regulator required for durotaxis. FASEB J. 32, 1099–1107 (2018).
Puleo, J. I. et al. Mechanosensing during directed cell migration requires dynamic actin polymerization at focal adhesions. J. Cell Biol. 218, 4215–4235 (2019).
Abercrombie, M. The Croonian Lecture, 1978—the crawling movement of metazoan cells. Proc. R. Soc. Lond. B 207, 129–147 (1980).
Mitchison, T. & Kirschner, M. Cytoskeletal dynamics and nerve growth. Neuron 1, 761–772 (1988).
Chan, C. E. & Odde, D. J. Traction dynamics of filopodia on compliant substrates. Science 322, 1687–1691 (2008).
Elosegui-Artola, A. et al. Mechanical regulation of a molecular clutch defines force transmission and transduction in response to matrix rigidity. Nat. Cell Biol. 18, 540–548 (2016).
DiMilla, P. A., Barbee, K. & Lauffenburger, D. A. Mathematical model for the effects of adhesion and mechanics on cell migration speed. Biophys. J. 60, 15–37 (1991).
Klank, R. L. et al. Biphasic dependence of glioma survival and cell migration on CD44 expression level. Cell Rep. 18, 23–31 (2017).
Rens, E. G. & Merks, R. M. H. Cell shape and durotaxis explained from cell-extracellular matrix forces and focal adhesion dynamics. iScience 23, 101488 (2020).
Schmidt, C. E., Dai, J., Lauffenburger, D. A., Sheetz, M. P. & Horwitz, A. F. Integrin-cytoskeletal interactions in neuronal growth cones. J. Neurosci. 15, 3400–3407 (1995).
Koser, D. E. et al. Mechanosensing is critical for axon growth in the developing brain. Nat. Neurosci. 19, 1592–1598 (2016).
Franze, K. Integrating chemistry and mechanics: the forces driving axon growth. Annu. Rev. Cell Dev. Biol. 36, 61–83 (2020).
Bangasser, B. L. & Odde, D. J. Master equation-based analysis of a motor-clutch model for cell traction force. Cell. Mol. Bioeng. 6, 449–459 (2013).
Bangasser, B. L., Rosenfeld, S. S. & Odde, D. J. Determinants of maximal force transmission in a motor-clutch model of cell traction in a compliant microenvironment. Biophys. J. 105, 581–592 (2013).
Cheng, B. et al. An integrated stochastic model of matrix-stiffness-dependent filopodial dynamics. Biophys. J. 111, 2051–2061 (2016).
Bangasser, B. L. et al. Shifting the optimal stiffness for cell migration. Nat. Commun. 8, 15313 (2017).
Lerche, M. et al. Integrin binding dynamics modulate ligand-specific mechanosensing in mammary gland fibroblasts. iScience 23, 100907 (2020).
Barber-Pérez, N. et al. Mechano-responsiveness of fibrillar adhesions on stiffness-gradient gels. J. Cell Sci. 133, jcs242909 (2020).
Miroshnikova, Y. A. et al. Tissue mechanics promote IDH1-dependent HIF1α–tenascin C feedback to regulate glioblastoma aggression. Nat. Cell Biol. 18, 1336–1345 (2016).
Kechagia, J. Z., Ivaska, J. & Roca-Cusachs, P. Integrins as biomechanical sensors of the microenvironment. Nat. Rev. Mol. Cell Biol. 20, 457–473 (2019).
Atherton, P. et al. Vinculin controls talin engagement with the actomyosin machinery. Nat. Commun. 6, 10038 (2015).
Cheng, B. et al. Nanoscale integrin cluster dynamics controls cellular mechanosensing via FAKY397 phosphorylation. Sci. Adv. 6, eaax1909 (2020).
Kanchanawong, P. et al. Nanoscale architecture of integrin-based cell adhesions. Nature 468, 580–584 (2010).
Mason, D. E. et al. YAP and TAZ limit cytoskeletal and focal adhesion maturation to enable persistent cell motility. J. Cell Biol. 218, 1369–1389 (2019).
Hou, J. C. et al. Modeling distributed forces within cell adhesions of varying size on continuous substrates. Cytoskeleton 76, 571–585 (2019).
Betz, T., Koch, D., Lu, Y.-B., Franze, K. & Käs, J. A. Growth cones as soft and weak force generators. Proc. Natl Acad. Sci. USA 108, 13420–13425 (2011).
Koch, D., Rosoff, W. J., Jiang, J., Geller, H. M. & Urbach, J. S. Strength in the periphery: growth cone biomechanics and substrate rigidity response in peripheral and central nervous system neurons. Biophys. J. 102, 452–460 (2012).
Ghibaudo, M. et al. Traction forces and rigidity sensing regulate cell functions. Soft Matter 4, 1836–1843 (2008).
Kuipers, A. J. et al. TRPM7 controls mesenchymal features of breast cancer cells by tensional regulation of SOX4. Biochim. Biophys. Acta Mol. Basis Dis. 1864, 2409–2419 (2018).
McGrail, D. J., Kieu, Q. M. N. & Dawson, M. R. The malignancy of metastatic ovarian cancer cells is increased on soft matrices through a mechanosensitive Rho–ROCK pathway. J. Cell Sci. 127, 2621–2626 (2014).
Fourriere, L. et al. RAB6 and microtubules restrict protein secretion to focal adhesions. J. Cell Biol. 218, 2215–2231 (2019).
Kobayashi, T. & Sokabe, M. Sensing substrate rigidity by mechanosensitive ion channels with stress fibers and focal adhesions. Curr. Opin. Cell Biol. 22, 669–676 (2010).
Wang, Y. L. & Pelham, R. J. Preparation of a flexible, porous polyacrylamide substrate for mechanical studies of cultured cells. Meth. Enzymol. 298, 489–496 (1998).
Polio, S. R. & Smith, M. L. Patterned hydrogels for simplified measurement of cell traction forces. Methods Cell. Biol. 121, 17–31 (2014).
Théry, M. & Piel, M. Adhesive micropatterns for cells: a microcontact printing protocol. Cold Spring Harb. Protoc. https://doi.org/10.1101/pdb.prot5255 (2009).
Han, S. J., Oak, Y., Groisman, A. & Danuser, G. Traction microscopy to identify force modulation in sub-resolution adhesions. Nat. Methods 12, 653–656 (2015).
Gillespie, D. T. Exact stochastic simulation of coupled chemical reactions. J. Phys. Chem. 81, 2340–2361 (1977).
We thank L. S. Prahl, J. Tian and G. Huang for helpful discussions on computational modelling and members of the Ivaska Lab for their insightful comments and discussion. Simulations were run in part on the high-performance computing resources at the Minnesota Supercomputing Institute. Turku Bioscience Centre Cell Imaging and Cytometry Core and Biocenter Finland are acknowledged for their services, instrumentation and expertise. We are supported by the University of Turku Doctoral Programme in Molecular Life Sciences (A.I.), the Company of Biologists Travelling Fellowship (A.I.), the Finnish Cultural Foundation (A.I.), the Academy of Finland (AoF CoE 346131 and 325464 (J.I.)), ERC CoG (grant 615258 (J.I.)), Sigrid Juselius Foundation (J.I.), the Finnish Cancer Organization (J.I.), the National Natural Science Foundation of China (11972280 (F.X.); 11772253 (M.L.); 12022206 (M.L.); 11532009 (T.J.L.); 12002262 (B.C.)), Natural Science Basic Research Plan in Shaanxi Province of China (2022KWZ-17 (M.L.)), the Shaanxi Province Youth Talent Support Program (M.L.), the Young Talent Support Plan of Xi’an Jiaotong University (M.L.), the National Institutes of Health (R01 AR077793 (G.M.G.); R01 CA172986 (D.J.O.); U54 CA210190 (D.J.O.); P01 CA254849 (D.J.O.); R35GM141853 (M.D.D.)) and the NSF Science and Technology Center for Engineering Mechanobiology (CMMI 1548571 (G.M.G.)).
The authors declare no competing interests.
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Supplementary Figs. 1–17, Notes 1–3, Tables 1–3, captions for Videos 1–3 and references.
Evolution of U-251MG glioblastoma cell distribution on photoresponsive stiffness gradient hydrogels over time.
Migration of individual U-251MG cells on photoresponsive stiffness gradient hydrogels.
DMSO- and H-1152-treated U-251MG cells migrating on stiffness gradients.
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Isomursu, A., Park, KY., Hou, J. et al. Directed cell migration towards softer environments. Nat. Mater. 21, 1081–1090 (2022). https://doi.org/10.1038/s41563-022-01294-2
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