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Actin as a Target to Reduce Cell Invasiveness in Initial Stages of Metastasis

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Abstract

We demonstrate the relative roles of the cell cytoskeleton, and specific importance of actin in facilitating mechanical aspects of metastatic invasion. A crucial step in metastasis, the typically lethal spread of cancer to distant body-sites, is cell invasion through dense tissues composed of extracellular matrix and various non-cancerous cells. Cell invasion requires cell-cytoskeleton remodeling to facilitate dynamic morphological changes and force application. We have previously shown invasive cell subsets in heterogeneous samples can rapidly (2 h) and forcefully indent non-degradable, impenetrable, synthetic gels to cell-scale depths. The amounts of indenting cells and their attained depths provide the mechanical invasiveness of the sample, which as we have shown agrees with the in vitro metastatic potential and the in vivo metastatic risk in humans. To identify invasive force-application mechanisms, we evaluated changes in mechanical invasiveness following chemical perturbations targeting the structure and function of cytoskeleton elements and associated proteins. We evaluate effects on short-term (2-hr) indentations of single, well-spaced or closely situated cells as compared to long-time-scale Boyden chamber migration. We show that actomyosin inhibition may be used to reduce (mechanical) invasiveness of single or collectively invading cells, while actin-disruption may induce escape-response of treated single-cells, which may promote metastasis.

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References

  1. Alvarez-Elizondo, M. B., C. W. Li, A. Marom, Y.-T. Tung, G. Drillich, Y. Horesh, S. C. Lin, G.-J. Wang, and D. Weihs. Micropatterned topographies reveal measurable differences between cancer and benign cells. Med. Eng. Phys. 75:5–12, 2020.

    PubMed  Google Scholar 

  2. Alvarez-Elizondo, M. B., and D. Weihs. Cell-gel mechanical interactions as an approach to rapidly and quantitatively reveal invasive subpopulations of metastatic cancer cells. Tissue Eng. Part C Methods 23:180–187, 2017.

    CAS  PubMed  Google Scholar 

  3. Blajeski, A. L., V. A. Phan, T. J. Kottke, and S. H. Kaufmann. G1 and G2 cell-cycle arrest following microtubule depolymerization in human breast cancer cells. J. Clin. Invest. 110:91–99, 2002.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Caswell, P. T., and T. Zech. Actin-based cell protrusion in a 3D matrix. Trends Cell Biol. 28:823–834, 2018.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Cheung, K. J., V. Padmanaban, V. Silvestri, K. Schipper, J. D. Cohen, A. N. Fairchild, M. A. Gorin, J. E. Verdone, K. J. Pienta, J. S. Bader, and A. J. Ewald. Polyclonal breast cancer metastases arise from collective dissemination of keratin 14-expressing tumor cell clusters. Proc. Natl. Acad. Sci. USA 113:E854–E863, 2016.

    CAS  PubMed  Google Scholar 

  6. Clark, A. G. A. G., and D. M. D. M. Vignjevic. Modes of cancer cell invasion and the role of the microenvironment. Curr. Opin. Cell Biol. 36:13–22, 2015.

    CAS  PubMed  Google Scholar 

  7. Dudaie, M., D. Weihs, F. J. Vermolen, and A. Gefen. Modeling migration in cell colonies in two and three dimensional substrates with varying stiffnesses. Silico Cell Tissue Sci. 2:1–14, 2015.

    Google Scholar 

  8. Dvir, L., R. Nissim, M. B. Alvarez-Elizondo, and D. Weihs. Quantitative measures to reveal coordinated cytoskeleton-nucleus reorganization during in vitro invasion of cancer cells. N. J. Phys. 17:043010, 2015.

    Google Scholar 

  9. Fidler, I. J. In vitro studies of cellular-mediated immunostimulation of tumor growth. J. Natl. Cancer Inst. 50:1307–1312, 1973.

    CAS  PubMed  Google Scholar 

  10. Fife, C. M., J. A. McCarroll, and M. Kavallaris. Movers and shakers: cell cytoskeleton in cancer metastasis. Br. J. Pharmacol. 171:5507–5523, 2014.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Friedl, P., and S. Alexander. Cancer invasion and the microenvironment: plasticity and reciprocity. Cell 147:992–1009, 2011.

    CAS  PubMed  Google Scholar 

  12. Friedl, P., J. Locker, E. Sahai, and J. E. Segall. Classifying collective cancer cell invasion. Nat. Cell Biol. 14:777–783, 2012.

    PubMed  Google Scholar 

  13. Gal, N., and D. Weihs. Intracellular mechanics and activity of breast cancer cells correlate with metastatic potential. Cell Biochem. Biophys. 63:199–209, 2012.

    CAS  PubMed  Google Scholar 

  14. Galbraith, C. G., K. M. Yamada, and M. P. Sheetz. The relationship between force and focal complex development. J. Cell Biol. 159:695–705, 2002.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Geiger, B., and K. M. Yamada. Molecular architecture and function of matrix adhesions. Cold Spring Harb. Perspect. Biol. 3:a005033, 2011.

    PubMed  PubMed Central  Google Scholar 

  16. Ghibaudo, M., A. Saez, L. Trichet, A. Xayaphoummine, J. Browaeys, P. Silberzan, A. Buguin, and B. Ladoux. Traction forces and rigidity sensing regulate cell functions. Soft Matter 4:1836–1843, 2008.

    CAS  Google Scholar 

  17. Gladilin, E., S. Ohse, M. Boerries, H. Busch, C. Xu, M. Schneider, M. Meister, and R. Eils. TGFβ-induced cytoskeletal remodeling mediates elevation of cell stiffness and invasiveness in NSCLC. Sci. Rep. 9:1–12, 2019.

    CAS  Google Scholar 

  18. Goldstein, D., T. Elhanan, M. Aronovitch, and D. Weihs. Origin of active transport in breast-cancer cells. Soft Matter 9:7167–7173, 2013.

    CAS  Google Scholar 

  19. Guck, J., S. Schinkinger, B. Lincoln, F. Wottawah, S. Ebert, M. Romeyke, D. Lenz, H. M. Erickson, R. Ananthakrishnan, D. Mitchell, J. Kas, S. Ulvick, and C. Bilby. Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. Biophys. J. 88:3689–3698, 2005.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Guzman, A., M. J. Ziperstein, and L. J. Kaufman. The effect of fibrillar matrix architecture on tumor cell invasion of physically challenging environments. Biomaterials 35:6954–6963, 2014.

    CAS  PubMed  Google Scholar 

  21. Haeger, A., S. Alexander, M. Vullings, F. M. P. Kaiser, C. Veelken, U. Flucke, G. E. Koehl, M. Hirschberg, M. Flentje, R. M. Hoffman, E. K. Geissler, S. Kissler, and P. Friedl. Collective cancer invasion forms an integrin-dependent radioresistant niche. J. Exp. Med. 217(1):e20181184, 2020.

    PubMed  Google Scholar 

  22. Isogai, T., R. van der Kammen, and M. Innocenti. SMIFH2 has effects on formins and p53 that perturb the cell cytoskeleton. Sci. Rep. 5:9802, 2015.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Khalil, A. A., O. Ilina, P. G. Gritsenko, P. Bult, P. N. Span, and P. Friedl. Collective invasion in ductal and lobular breast cancer associates with distant metastasis. Clin. Exp. Metastasis 34:421–429, 2017.

    PubMed  PubMed Central  Google Scholar 

  24. Kraning-Rush, C. M., J. P. Califano, and C. A. Reinhart-King. Cellular traction stresses increase with increasing metastatic potential. PLoS ONE 7:e32572, 2012.

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Kristal-Muscal, R., L. Dvir, and D. Weihs. Metastatic cancer cells tenaciously indent impenetrable, soft substrates. N. J. Phys. 15:035022, 2013.

    CAS  Google Scholar 

  26. Li, Q., Z. Ma, Y. Liu, X. Kan, C. Wang, B. Su, Y. Li, Y. Zhang, P. Wang, Y. Luo, D. Na, L. Wang, G. Zhang, X. Zhu, and L. Wang. Low doses of paclitaxel enhance liver metastasis of breast cancer cells in the mouse model. FEBS J. 283:2836–2852, 2016.

    CAS  PubMed  Google Scholar 

  27. Mak, M., C. A. Reinhart-King, and D. Erickson. Elucidating mechanical transition effects of invading cancer cells with a subnucleus-scaled microfluidic serial dimensional modulation device. Lab Chip 13:340–348, 2013.

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Massalha, S., and D. Weihs. Metastatic breast cancer cells adhere strongly on varying stiffness substrates, initially without adjusting their morphology. Biomech. Model. Mechanobiol. 16:961–970, 2017.

    PubMed  Google Scholar 

  29. Merkher, Y., M. B. Alvarez-Elizondo, and D. Weihs. Taxol reduces synergistic, mechanobiological invasiveness of metastatic cells. Converg. Sci. Phys. Oncol. 3:044002, 2017.

    Google Scholar 

  30. Merkher, Y., Y. Horesh, Z. Abramov, G. Shleifer, O. Ben-Ishay, Y. Kluger, and D. Weihs. Rapid cancer diagnosis and early prognosis of metastatic risk based on mechanical invasiveness of sampled cells. Ann. Biomed. Eng. 2020. https://doi.org/10.1007/s10439-020-02547-4.

    Article  PubMed  Google Scholar 

  31. Merkher, Y., and D. Weihs. Proximity of metastatic cells enhances their mechanobiological invasiveness. Ann. Biomed. Eng. 45:1399–1406, 2017.

    PubMed  Google Scholar 

  32. Mierke, C. T. The biomechanical properties of 3d extracellular matrices and embedded cells regulate the invasiveness of cancer cells. Cell Biochem. Biophys. 61:217–236, 2011.

    CAS  PubMed  Google Scholar 

  33. Mierke, C. T., B. Frey, M. Fellner, M. Herrmann, and B. Fabry. Integrin alpha5beta1 facilitates cancer cell invasion through enhanced contractile forces. J. Cell Sci. 124:369–383, 2011.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Mohammadi, A., B. Mansoori, M. Aghapour, S. Shirjang, S. Nami, and B. Baradaran. The urtica dioica extract enhances sensitivity of paclitaxel drug to MDA-MB-468 breast cancer cells. Biomed. Pharmacother. 83:835–842, 2016.

    CAS  PubMed  Google Scholar 

  35. O’Shaughnessy, J., W. J. Gradishar, P. Bhar, and J. Iglesias. nab-Paclitaxel for first-line treatment of patients with metastatic breast cancer and poor prognostic factors: a retrospective analysis. Breast Cancer Res. Treat. 138:829–837, 2013.

    PubMed  PubMed Central  Google Scholar 

  36. Patsialou, A., J. J. Bravo-Cordero, Y. Wang, D. Entenberg, H. Liu, M. Clarke, and J. S. Condeelis. Intravital multiphoton imaging reveals multicellular streaming as a crucial component of in vivo cell migration in human breast tumors. Intravital 2:e25294, 2013.

    PubMed  PubMed Central  Google Scholar 

  37. Peterson, J. A., B. Tian, A. D. Bershadsky, T. Volberg, R. E. Gangnon, I. Spector, B. Geiger, and P. L. Kaufman. Latrunculin-A increases outflow facility in the monkey. Investig. Ophthalmol. Vis. Sci. 40:931–941, 1999.

    CAS  Google Scholar 

  38. Poincloux, R., O. Collin, F. Lizarraga, M. Romao, M. Debray, M. Piel, and P. Chavrier. Contractility of the cell rear drives invasion of breast tumor cells in 3D matrigel. Proc. Natl. Acad. Sci. USA 108:1943–1948, 2011.

    CAS  PubMed  Google Scholar 

  39. Seetharaman, S., and S. Etienne-Manneville. Cytoskeletal crosstalk in cell migration. Trends Cell Biol. 30:720–735, 2020.

    CAS  PubMed  Google Scholar 

  40. Sen, S., A. J. Engler, and D. E. Discher. Matrix strains induced by cells: computing how far cells can feel. Cell. Mol. Bioeng. 2:39–48, 2009.

    PubMed  PubMed Central  Google Scholar 

  41. Stearns, M. E., and M. Wang. Taxol blocks processes essential for prostate tumor cell (PC-3 ML) invasion and metastases. Cancer Res. 52:3776–3781, 1992.

    CAS  PubMed  Google Scholar 

  42. Stehn, J. R., N. K. Haass, T. Bonello, M. Desouza, G. Kottyan, H. Treutlein, J. Zeng, P. R. B. B. Nascimento, V. B. Sequeira, T. L. Butler, M. Allanson, T. Fath, T. A. Hill, A. McCluskey, G. Schevzov, S. J. Palmer, E. C. Hardeman, D. Winlaw, V. E. Reeve, I. Dixon, W. Weninger, T. P. Cripe, and P. W. Gunning. A novel class of anticancer compounds targets the actin cytoskeleton in tumor cells. Cancer Res. 73:5169–5182, 2013.

    CAS  PubMed  Google Scholar 

  43. Swaminathan, V., K. Mythreye, E. T. O’Brien, A. Berchuck, G. C. Blobe, and R. Superfine. Mechanical stiffness grades metastatic potential in patient tumor cells and in cancer cell lines. Cancer Res. 71:5075–5080, 2011.

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Tozluoglu, M., A. L. Tournier, R. P. Jenkins, S. Hooper, P. A. Bates, and E. Sahai. Matrix geometry determines optimal cancer cell migration strategy and modulates response to interventions. Nat. Cell Biol. 15:751–762, 2013.

    CAS  PubMed  Google Scholar 

  45. Volk-Draper, L., K. Hall, C. Griggs, S. Rajput, P. Kohio, D. DeNardo, and S. Ran. Paclitaxel therapy promotes breast cancer metastasis in a TLR4-dependent manner. Cancer Res. 74:5421–5434, 2014.

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Wang, N., and D. E. Ingber. Probing transmembrane mechanical coupling and cytomechanics using magnetic twisting cytometry. Biochem. Cell Biol. 73:327–335, 1995.

    CAS  PubMed  Google Scholar 

  47. Weihs, D., Y. Merkher. A device and method for determining cell indention activity, Patent pending. Patent: PCT/IL2019/050463, 2019.

  48. Weihs, D., T. G. Mason, and M. A. Teitell. Effects of cytoskeletal disruption on transport, structure, and rheology within mammalian cells. Phys. Fluids 19:103102, 2007.

    Google Scholar 

  49. Wyckoff, J. B., S. E. Pinner, S. Gschmeissner, J. S. Condeelis, and E. Sahai. ROCK- and myosin-dependent matrix deformation enables protease-independent tumor-cell invasion in vivo. Curr. Biol. 16:1515–1523, 2006.

    CAS  PubMed  Google Scholar 

  50. Wyse, M. M., J. Lei, A. L. Nestor-Kalinoski, and K. M. Eisenmann. Dia-interacting protein (DIP) imposes migratory plasticity in mDia2-dependent tumor cells in three-dimensional matrices. PLoS ONE 7:e45085, 2012.

    CAS  PubMed  PubMed Central  Google Scholar 

  51. Yeung, T., P. C. Georges, L. A. Flanagan, B. Marg, M. Ortiz, M. Funaki, N. Zahir, W. Ming, V. Weaver, and P. A. Janmey. Effects of substrate stiffness on cell morphology, cytoskeletal structure, and adhesion. Cell Motil. Cytoskeleton 60:24–34, 2005.

    PubMed  Google Scholar 

  52. Yilmaz, M., and G. Christofori. Mechanisms of motility in metastasizing cells. Mol. Cancer Res. 8:629–642, 2010.

    CAS  PubMed  Google Scholar 

  53. Yizraeli, M. L., and D. Weihs. Time-dependent micromechanical responses of breast cancer cells and adjacent fibroblasts to electric treatment. Cell Biochem. Biophys. 61:605–618, 2011.

    CAS  PubMed  Google Scholar 

  54. Zhang, Y., Y. Wang, and J. Xue. Paclitaxel inhibits breast cancer metastasis via suppression of aurora kinase-mediated cofilin-1 activity. Exp. Ther. Med. 15:1269–1276, 2018.

    CAS  PubMed  Google Scholar 

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Acknowledgments

The authors thank Mr. Yam Horesh for his assistance with the cell viability determination. The work was partially funded by the Elias Fund for Medical Research, the Polak Fund for Applied Research, and the Samuel H. Born Fund for Biomedical Research.

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No benefits in any form have been or will be received from a commercial party related directly or indirectly to the subject of this manuscript.

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Author notes

  1. Martha B. Alvarez-Elizondo and Yulia Merkher have contributed equally to this work.

Authors and Affiliations

  1. Faculty of Biomedical Engineering, Technion-Israel Institute of Technology, Haifa, 3200003, Israel

    Martha B. Alvarez-Elizondo, Yulia Merkher, Gal Shleifer, Carmel Gashri & Daphne Weihs

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  1. Martha B. Alvarez-Elizondo
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Correspondence to Daphne Weihs.

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Alvarez-Elizondo, M.B., Merkher, Y., Shleifer, G. et al. Actin as a Target to Reduce Cell Invasiveness in Initial Stages of Metastasis. Ann Biomed Eng 49, 1342–1352 (2021). https://doi.org/10.1007/s10439-020-02679-7

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