Fluidic substrate as a tool to probe breast cancer cell adaptive behavior in response to fluidity level

Abstract

Although the roles of elastic components in breast cancer progression have been widely studied, the importance of matrix dissipative elements in regulating breast cancer behavior is still poorly understood. In this study, we designed viscosity-tunable fluidic substrates to investigate the effects of matrix viscosity on the alteration of breast cancer cellular fate using a hydrophobic molten polymer of poly(ε-caprolactone-co-D,L-lactide) [P(CL-co-DLLA)] with different levels of fluidity. The high- and low-fluidity substrates used in this study were shown to behave as viscoelastic liquids at physiological temperature. A nonmetastatic breast cancer cell line (MCF-7) was cultured at the interface of the fibronectin-coated substrate, and its behavior towards the substrate fluidity level was thoroughly characterized. Despite fibronectin-mediated cell-substrate interactions, MCF-7 cells show sensitivity to substrate fluidity levels by forming types aggregates of different sizes and structures over time. Accordingly, MCF-7 cells were undergoing senescence on fluidic substrates, as shown by high metabolic activity over time, suppressed proliferation ability, and positive expression of senescence markers. Moreover, senescence implies more resistance towards anticancer drug treatment. This indicates that a fluidic substrate, as a two-dimensional synthetic matrix system, could demonstrate the importance of mechanical cues in redefining cellular function and cellular fate by changing the viscosity of the pure substrate.

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References

  1. 1.

    Oskarsson T. Extracellular matrix components in breast cancer progression and metastasis. Breast. 2013;22:566–72. https://doi.org/10.1016/j.breast.2013.07.012.

    Article  Google Scholar 

  2. 2.

    Lee J, Chaudhuri O. Regulation of breast cancer progression by extracellular matrix mechanics: insights from 3D culture models. ACS Biomater Sci Eng.2018;4:302–13. https://doi.org/10.1021/acsbiomaterials.7b00071.

    CAS  Article  Google Scholar 

  3. 3.

    Papalazarou V, Salmeron-Sanchez M, Machesky LM. Tissue engineering the cancer microenvironment-challenges and opportunities. Biophys Rev. 2018;10:1695–711. https://doi.org/10.1007/s12551-018-0466-8.

    Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Walker C, Mojares E, del Río Hernández A. Role of extracellular matrix in development and cancer progression. Int J Mol Sci. 2018;19:3028 https://doi.org/10.3390/ijms19103028.

    CAS  Article  PubMed Central  Google Scholar 

  5. 5.

    Muncie JM, Weaver VM. The physical and biochemical properties of the extracellular matrix regulate cell fate. Curr Top Dev Biol. 2018;130:1–37. https://doi.org/10.1016/bs.ctdb.2018.02.002.

    Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Frantz C, Stewart KM, Weaver VM. The extracellular matrix at a glance. J Cell Sci. 2010;123:4195–200. https://doi.org/10.1242/jcs.023820.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Devi CU, Chandran RB, Vasu RM, Sood AK. Measurement of visco-elastic properties of breast-tissue mimicking materials using diffusing wave spectroscopy. J Biomed Opt. 2007;12:034035 https://doi.org/10.1117/1.2743081.

    Article  PubMed  Google Scholar 

  8. 8.

    Chaudhuri PK, Low BC, Lim CT. Mechanobiology of tumor growth. Chem Rev. 2018;118:6499–515. https://doi.org/10.1021/acs.chemrev.8b00042.

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Charrier EE, Pogoda K, Wells RG, Janmey PA. Control of cell morphology and differentiation by substrates with independently tunable elasticity and viscous dissipation. Nat Commun. 2018;9:449 https://doi.org/10.1038/s41467-018-02906-9.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  10. 10.

    Murrell M, Kamm R, Matsudaira P. Substrate viscosity enhances correlation in epithelial sheet movement. Biophys J. 2011;101:297–306. https://doi.org/10.1016/j.bpj.2011.05.048.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Gonzalez-Molina J, Zhang X, Borghesan M, da Silva JM, Awan M, Fuller B. et al. Extracellular fluid viscosity enhances liver cancer cell mechanosensing and migration. Biomaterials . 2018;177:113–24. https://doi.org/10.1016/j.biomaterials.2018.05.058.

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Cantini M, Donnelly H, Dalby MJ, Salmeron‐Sanchez M. The plot thickens: the emerging role of matrix viscosity in cell mechanotransduction. Adv Healthcare Mater. 2019. https://doi.org/10.1002/adhm.201901259.

  13. 13.

    Poh PSP, Hege C, Chhaya MP, Balmayor ER, Foehr P, Burgkart RH, et al. Evaluation of polycaprolactone−poly-D,L-lactide copolymer as biomaterial for breast tissue engineering. Polym Int. 2017;66:77–84. https://doi.org/10.1002/pi.5181.

    CAS  Article  Google Scholar 

  14. 14.

    Mano SS, Uto K, Aoyagi T, Ebara M. Fluidity of biodegradable substrate regulates carcinoma cell behavior: a novel approach to cancer therapy. AIMS Mater Sci. 2016;3:66–82. https://doi.org/10.3934/matersci.2016.1.66.

    CAS  Article  Google Scholar 

  15. 15.

    Mano SS, Uto K, Ebara M. Material-induced senescence (MIS): fluidity induces senescent type cell death of lung cancer cells via insulin-like growth factor binding protein 5. Theranostics. 2017;7:4658–70. https://doi.org/10.7150/thno.20582.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Uto K, Mano SS, Aoyagi T, Ebara M. Substrate fluidity regulates cell adhesion and morphology on poly (ε-caprolactone)-based materials. ACS Biomater Sci Eng. 2016;2:446–53. https://doi.org/10.1021/acsbiomaterials.6b00058.

    CAS  Article  Google Scholar 

  17. 17.

    Uto K, Muroya T, Okamoto M, Tanaka H, Murase T, Ebara M, et al. Design of super-elastic biodegradable scaffolds with longitudinally oriented microchannels and optimization of the channel size for Schwann cell migration. Sci Technol Adv Mater. 2012;13:064207. https://doi.org/10.1088/1468-6996/13/6/064207.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Dzhoyashvili NA, Thompson K, Gorelov AV, Rochev YA. Film thickness determines cell growth and cell sheet detachment from spin-coated poly(N‑Isopropylacrylamide) substrates. ACS Appl Mater Interfaces. 2016;8:27564–572. https://doi.org/10.1021/acsami.6b09711.

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    Buxboim A, Rajagopal K, Brown AE, Discher DE. How deeply cells feel: methods for thin gels. J Condens. 2010;22:194116. https://doi.org/10.1088/0953-8984/22/19/194116.

    CAS  Article  Google Scholar 

  20. 20.

    Chaudhuri PK, Pan CQ, Low BC, Lim CT. Differential depth sensing reduces cancer cell proliferation via rho-rac-regulated invadopodia. ACS Nano. 2017;11:7336–48. https://doi.org/10.1021/acsnano.7b03452.

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Chester D, Kathard R, Nortey J, Nellenbach K, Brown AC. Viscoelastic properties of microgel thin films control fibroblast modes of migration and pro-fibrotic responses. Biomaterials. 2018;185:371–82. https://doi.org/10.1016/j.biomaterials.2018.09.012.

    CAS  Article  PubMed  Google Scholar 

  22. 22.

    Bennett M, Cantini M, Reboud J, Cooper JM, Roca-Cusachs P, Salmeron-Sanchez M. Molecular clutch drives cell response to surface viscosity. PNAS. 2018;115:1192–7. https://doi.org/10.1073/pnas.1710653115.

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Kourouklis AP, Lerum RV, Bermudez H. Cell adhesion mechanisms on laterally mobile polymer films. Biomaterials. 2014;35:4827–34. https://doi.org/10.1016/j.biomaterials.2014.02.052.

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Zheng JY, Han SP, Chiu YJ, Yip AK, Boichat N, Zhu SW, et al. Epithelial monolayers coalesce on a viscoelastic substrate through redistribution of vinculin. Biophys J. 2017;113:1585–98. https://doi.org/10.1016/j.bpj.2017.07.027.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Bell S, Terentjev EM. Focal adhesion kinase: the reversible molecular mechanosensor. Biophys J. 2017;112:2439–50. https://doi.org/10.1016/j.bpj.2017.04.048.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  26. 26.

    Gorgoulis V, Adams PD, Alimonti A, Bennett DC, Bischof O, Bishop C, et al. Cellular senescence: defining a path forward. Cell. 2019;179:813–27. https://doi.org/10.1016/j.cell.2019.10.005.

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Zhang W, Choi DS, Nguyen YH, Chang J, Qin L. Studying cancer stem cell dynamics on PDMS surfaces for microfluidics device design. Sci Rep. 2013;3:2332. https://doi.org/10.1038/srep02332.

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors would like to express their gratitude to the Grants-in-Aid for Scientific Research (B) (19H04476) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) Japan. The authors are grateful to Allan S. Hoffman (University of Washington) for continued and valuable discussion.

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Correspondence to Mitsuhiro Ebara.

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Najmina, M., Uto, K. & Ebara, M. Fluidic substrate as a tool to probe breast cancer cell adaptive behavior in response to fluidity level. Polym J 52, 985–995 (2020). https://doi.org/10.1038/s41428-020-0345-6

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