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Enhanced substrate stress relaxation promotes filopodia-mediated cell migration

Nature Materials volume 20, pages 1290–1299 (2021)Cite this article

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Abstract

Cell migration on two-dimensional substrates is typically characterized by lamellipodia at the leading edge, mature focal adhesions and spread morphologies. These observations result from adherent cell migration studies on stiff, elastic substrates, because most cells do not migrate on soft, elastic substrates. However, many biological tissues are soft and viscoelastic, exhibiting stress relaxation over time in response to a deformation. Here, we have systematically investigated the impact of substrate stress relaxation on cell migration on soft substrates. We observed that cells migrate minimally on substrates with an elastic modulus of 2 kPa that are elastic or exhibit slow stress relaxation, but migrate robustly on 2-kPa substrates that exhibit fast stress relaxation. Strikingly, migrating cells were not spread out and did not extend lamellipodial protrusions, but were instead rounded, with filopodia protrusions extending at the leading edge, and exhibited small nascent adhesions. Computational models of cell migration based on a motor–clutch framework predict the observed impact of substrate stress relaxation on cell migration and filopodia dynamics. Our findings establish substrate stress relaxation as a key requirement for robust cell migration on soft substrates and uncover a mode of two-dimensional cell migration marked by round morphologies, filopodia protrusions and weak adhesions.

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Fig. 1: Substrate stress relaxation regulates cell migration on soft substrates.
Fig. 2: Cells migrate on viscoelastic substrates with rounded morphologies.
Fig. 3: Filopodia protrusions mediate migration on soft, fast-relaxing substrates.
Fig. 4: Adhesion and traction force mediate migration on fast-relaxing substrates.
Fig. 5: Simulations capture the impact of substrate stress relaxation on cell migration.
Fig. 6: Substrate stress relaxation regulates filopodia dynamics.

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Data availability

All data relevant to this manuscript are available upon request. Source data are provided with this paper.

Code availability

All analyses codes relevant to this manuscript have been deposited in the DOI-minting repository Zenodo54. Simulation codes are available upon request.

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Acknowledgements

We acknowledge R. Stowers (University of California, Santa Barbara) and the Chaudhuri laboratory for helpful discussion, and M. Levenston (Stanford University) for use of mechanical testing equipment. We also acknowledge the Stanford Cell Sciences Imaging Facility for Imaris software access and for technical assistance with Imaris. Figure 5n is a schematic created with BioRender.com. K.A. acknowledges financial support from the Stanford ChEM-H Chemistry/Biology Interface Predoctoral Training Program and the National Institute of General Medical Sciences of the National Institutes of Health under Award Number T32GM120007, and a National Science Foundation (NSF) Graduate Student fellowship. D.G. was funded in part by a National Institutes of Health Fellowship under Award Number GM116328. This work was supported by a National Institutes of Health National Cancer Institute (NIH NCI) grant (U54 CA210190) for D.J.O. and NIH NCI grant R01 CA232256, NIH National Institute of Biomedical Imaging and Bioengineering awards R01EB017753 and R01EB030876, NSF Center for Engineering Mechanobiology grant CMMI-154857, and NSF grants MRSEC/DMR-1720530 and DMS-1953572 to V.B.S., and by an American Cancer Society grant (RSG-16-208-01) and a NIH NCI grant (R37 CA214136) to O.C.

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Authors and Affiliations

  1. Department of Chemical Engineering, Stanford University, Stanford, CA, USA

    Kolade Adebowale

  2. Chemistry, Engineering, and Medicine for Human Health (ChEM-H), Stanford University, Stanford, CA, USA

    Kolade Adebowale & Ovijit Chaudhuri

  3. Department of Materials Science and Engineering, University of Pennsylvania, Philadelphia, PA, USA

    Ze Gong & Vivek B. Shenoy

  4. Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA

    Jay C. Hou & David J. Odde

  5. Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA

    Katrina M. Wisdom & Vivek B. Shenoy

  6. Department of Mechanical Engineering, Stanford University, Stanford, CA, USA

    Katrina M. Wisdom, Hong-pyo Lee, Sungmin Nam & Ovijit Chaudhuri

  7. Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA, USA

    Damien Garbett & Tobias Meyer

  8. John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, USA

    Sungmin Nam

  9. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA

    Sungmin Nam

  10. Department of Cell and Developmental Biology, Weill Cornell Medical College, New York, NY, USA

    Tobias Meyer

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Contributions

K.A. and O.C. designed the experiments. K.M.W. designed the material system. K.A. performed rheometry, substrate preparation, time-lapse microscopy cell migration, immunofluorescence and filopodia experiments. H.L. performed the calcium imaging experiments. S.N. helped with the traction force experiments. D.G. and T.M. performed the siRNA knockdown experiments. K.A. performed all experimental data analysis and statistical tests. Z.G., V.B.S., J.C.H. and D.J.O. performed the computer simulations and analysis. K.A., Z.G., J.C.H., V.B.S., D.J.O. and O.C. wrote the manuscript.

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Correspondence to Ovijit Chaudhuri.

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Peer review Information Nature Materials thanks Pakorn Kanchanawong and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–8, Tables 1–4, Videos 1–5, notes and references.

Reporting Summary

Supplementary Video 1

Top view of HT-1080 cells migrating on fast-relaxing substrates. Scale bar, 20 μm. Time, h:min.

Supplementary Video 2

Top view of HT-1080 cells migrating on slow-relaxing substrates. Scale bar, 20 μm. Time, h:min.

Supplementary Video 3

Top view of HT-1080 cell tugging on substrate and subsequent release (yellow square). Green, cell; red, fiducial marker. Scale bar, 20 μm. Time, h:min.

Supplementary Video 4

Live imaging of HT-1080 cells (actin-labelled) filopodia on fast-relaxing substrates. Scale bar, 20 μm. Time, h:min.

Supplementary Video 5

Live imaging of HT-1080 cells (actin-labelled) filopodia on slow-relaxing substrates. Scale bar, 20 μm. Time, h:min.

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Adebowale, K., Gong, Z., Hou, J.C. et al. Enhanced substrate stress relaxation promotes filopodia-mediated cell migration. Nat. Mater. 20, 1290–1299 (2021). https://doi.org/10.1038/s41563-021-00981-w

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