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Analysis of Energy-Driven Leader-Follower Hierarchy During Collective Cancer Cell Invasion

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Cell Migration in Three Dimensions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2608))

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Abstract

Many solid tumors can invade the surrounding three-dimensional (3D) tissue in a collective manner, and increasing evidence suggests that collective migration makes cancer cell clusters more invasive and metastatic than individual cells. A cohesive cohort of cancer cells can have many advantages over individual cells, including more efficient bioenergetics that have been recently identified. Minimization of bioenergetic costs during collective cell migration drives leader-follower dynamics and contributes to enhanced cancer invasion. Hence, it is critical to understand the migratory and bioenergetic dynamics of cancer collective invasion. While analysis of structures and dynamics in a 3D space has been a challenging task, here we describe a widely applicable method to analyze the energy-driven leader-follower hierarchy during cancer collective invasion. An in vitro tumor spheroid model is employed to reproduce the in vivo collective behaviors of cancer cells while allowing high spatiotemporal resolution imaging, where the leader-follower dynamics can be analyzed by tracking nuclear positions. As glucose is one of the main energy sources that fuel cancer cell migration, the quantification of glucose uptake along the invading strands provides an estimate of the energy demand associated with collective invasion. Finally, we describe a method to quantify the dynamics of intracellular energy level using the PercevalHR ATP:ADP ratio biosensor.

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References

  1. Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100(1):57–70. https://doi.org/10.1016/s0092-8674(00)81683-9

    Article  CAS  Google Scholar 

  2. Thiery JP (2002) Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer 2(6):442–454. https://doi.org/10.1038/nrc822

    Article  CAS  Google Scholar 

  3. Friedl P, Locker J, Sahai E, Segall JE (2012) Classifying collective cancer cell invasion. Nat Cell Biol 14(8):777–783. https://doi.org/10.1038/ncb2548

    Article  CAS  Google Scholar 

  4. Cheung KJ, Ewald AJ (2016) A collective route to metastasis: seeding by tumor cell clusters. Science 352(6282):167–169. https://doi.org/10.1126/science.aaf6546

    Article  CAS  Google Scholar 

  5. Lang C, Conrad L, Iber D (2021) Organ-specific branching morphogenesis. Front Cell Dev Biol 9:671402. https://doi.org/10.3389/fcell.2021.671402

    Article  Google Scholar 

  6. Bordeleau F, Mason BN, Lollis EM, Mazzola M, Zanotelli MR, Somasegar S, Califano JP, Montague C, LaValley DJ, Huynh J, Mencia-Trinchant N, Negron Abril YL, Hassane DC, Bonassar LJ, Butcher JT, Weiss RS, Reinhart-King CA (2017) Matrix stiffening promotes a tumor vasculature phenotype. Proc Natl Acad Sci U S A 114(3):492–497. https://doi.org/10.1073/pnas.1613855114

    Article  CAS  Google Scholar 

  7. Zahm J-M, Kaplan H, Hérard A-L, Doriot F, Pierrot D, Somelette P, Puchelle E (1997) Cell migration and proliferation during the in vitro wound repair of the respiratory epithelium. Cell Motil Cytoskeleton 37(1):33–43. https://doi.org/10.1002/(sici)1097-0169(1997)37:1<33::Aid-cm4>3.0.Co;2-i

    Article  CAS  Google Scholar 

  8. Mark C, Grundy TJ, Strissel PL, Bohringer D, Grummel N, Gerum R, Steinwachs J, Hack CC, Beckmann MW, Eckstein M, Strick R, O’Neill GM, Fabry B (2020) Collective forces of tumor spheroids in three-dimensional biopolymer networks. elife 9. https://doi.org/10.7554/eLife.51912

  9. Ellison D, Mugler A, Brennan MD, Lee SH, Huebner RJ, Shamir ER, Woo LA, Kim J, Amar P, Nemenman I, Ewald AJ, Levchenko A (2016) Cell-cell communication enhances the capacity of cell ensembles to sense shallow gradients during morphogenesis. Proc Natl Acad Sci U S A 113(6):E679–E688. https://doi.org/10.1073/pnas.1516503113

    Article  CAS  Google Scholar 

  10. Sunyer R, Conte V, Escribano J, Elosegui-Artola A, Labernadie A, Valon L, Navajas D, Garcia-Aznar JM, Munoz JJ, Roca-Cusachs P, Trepat X (2016) Collective cell durotaxis emerges from long-range intercellular force transmission. Science 353(6304):1157–1161. https://doi.org/10.1126/science.aaf7119

    Article  CAS  Google Scholar 

  11. Zhang J, Goliwas KF, Wang W, Taufalele PV, Bordeleau F, Reinhart-King CA (2019) Energetic regulation of coordinated leader-follower dynamics during collective invasion of breast cancer cells. Proc Natl Acad Sci U S A 116(16):7867–7872. https://doi.org/10.1073/pnas.1809964116

    Article  CAS  Google Scholar 

  12. Liu L, Duclos G, Sun B, Lee J, Wu A, Kam Y, Sontag ED, Stone HA, Sturm JC, Gatenby RA, Austin RH (2013) Minimization of thermodynamic costs in cancer cell invasion. Proc Natl Acad Sci U S A 110(5):1686–1691. https://doi.org/10.1073/pnas.1221147110

    Article  Google Scholar 

  13. Zanotelli MR, Zhang J, Reinhart-King CA (2021) Mechanoresponsive metabolism in cancer cell migration and metastasis. Cell Metab 33:1307. https://doi.org/10.1016/j.cmet.2021.04.002

    Article  CAS  Google Scholar 

  14. Zanotelli MR, Goldblatt ZE, Miller JP, Bordeleau F, Li J, VanderBurgh JA, Lampi MC, King MR, Reinhart-King CA (2018) Regulation of ATP utilization during metastatic cell migration by collagen architecture. Mol Biol Cell 29(1):1–9. https://doi.org/10.1091/mbc.E17-01-0041

    Article  CAS  Google Scholar 

  15. Wu Y, Zanotelli MR, Zhang J, Reinhart-King CA (2021) Matrix-driven changes in metabolism support cytoskeletal activity to promote cell migration. Biophys J 120:1705. https://doi.org/10.1016/j.bpj.2021.02.044

    Article  CAS  Google Scholar 

  16. Commander R, Wei C, Sharma A, Mouw JK, Burton LJ, Summerbell E, Mahboubi D, Peterson RJ, Konen J, Zhou W, Du Y, Fu H, Shanmugam M, Marcus AI (2020) Subpopulation targeting of pyruvate dehydrogenase and GLUT1 decouples metabolic heterogeneity during collective cancer cell invasion. Nat Commun 11(1):1533. https://doi.org/10.1038/s41467-020-15219-7

    Article  CAS  Google Scholar 

  17. Tantama M, Martinez-Francois JR, Mongeon R, Yellen G (2013) Imaging energy status in live cells with a fluorescent biosensor of the intracellular ATP-to-ADP ratio. Nat Commun 4:2550. https://doi.org/10.1038/ncomms3550

    Article  CAS  Google Scholar 

  18. Hung YP, Albeck JG, Tantama M, Yellen G (2011) Imaging cytosolic NADH-NAD(+) redox state with a genetically encoded fluorescent biosensor. Cell Metab 14(4):545–554. https://doi.org/10.1016/j.cmet.2011.08.012

    Article  CAS  Google Scholar 

  19. Takanaga H, Chaudhuri B, Frommer WB (2008) GLUT1 and GLUT9 as major contributors to glucose influx in HepG2 cells identified by a high sensitivity intramolecular FRET glucose sensor. Biochim Biophys Acta 1778(4):1091–1099. https://doi.org/10.1016/j.bbamem.2007.11.015

    Article  CAS  Google Scholar 

  20. Tantama M, Hung YP, Yellen G (2011) Imaging intracellular pH in live cells with a genetically encoded red fluorescent protein sensor. J Am Chem Soc 133(26):10034–10037. https://doi.org/10.1021/ja202902d

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by funding from the National Institutes of Health (HL127499 and GM13117) and the W.M. Keck Foundation to C.A.R.-K.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, USA

    Jian Zhang & Cynthia A. Reinhart-King

Authors
  1. Jian Zhang
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  2. Cynthia A. Reinhart-King
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Corresponding author

Correspondence to Cynthia A. Reinhart-King .

Editor information

Editors and Affiliations

  1. Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, Amsterdam, The Netherlands

    Coert Margadant

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Zhang, J., Reinhart-King, C.A. (2023). Analysis of Energy-Driven Leader-Follower Hierarchy During Collective Cancer Cell Invasion. In: Margadant, C. (eds) Cell Migration in Three Dimensions. Methods in Molecular Biology, vol 2608. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2887-4_15

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  • DOI: https://doi.org/10.1007/978-1-0716-2887-4_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2886-7

  • Online ISBN: 978-1-0716-2887-4

  • eBook Packages: Springer Protocols

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