Abstract
Cellular traction forces are contractile forces that depend on the material/substrate stiffness and play essential roles in sensing mechanical environments and regulating cell morphology and function. Traction forces are primarily generated by the actin cytoskeleton and transmitted to the substrate through focal adhesions. The cell nucleus is also believed to be involved in the regulation of this type of force; however, the role of the nucleus in cellular traction forces remains unclear. In this study, we explored the effects of nucleus-actin filament coupling on cellular traction forces in human dermal fibroblasts cultured on substrates with varying stiffness (5, 15, and 48 kPa). To investigate these effects, we transfected the cells with a dominant-negative Klarsicht/ANC-1/Syne homology (DN-KASH) protein that was designed to displace endogenous linker proteins and disrupt nucleus-actin cytoskeleton connections. The force that exists between the cytoskeleton and the nucleus (nuclear tension) was also evaluated with a fluorescence resonance energy transfer (FRET)-based tension sensor. We observed a biphasic change in cellular traction forces with a peak at 15 kPa, regardless of DN-KASH expression, that was inversely correlated with the nuclear tension. In addition, the relative magnitude and distribution of traction forces in nontreated wild-type cells were similar across different stiffness conditions, while DN-KASH-transfected cells exhibited a different distribution pattern that was impacted by the substrate stiffness. These results suggest that the nucleus-actin filament coupling play a homeostatic role by maintaining the relative magnitude of cellular traction forces in fibroblasts under different stiffness conditions.
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No datasets were generated or analysed during the current study.
References
Achterberg VF et al (2014) The nano-scale mechanical properties of the extracellular matrix regulate dermal fibroblast function. J Invest Dermatol 134:1862–1872. https://doi.org/10.1038/jid.2014.90
Ambrosi D, Duperray A, Peschetola V, Verdier C (2009) Traction patterns of tumor cells. J Math Biol 58:163–181. https://doi.org/10.1007/s00285-008-0167-1
Anno T, Sakamoto N, Sato M (2012) Role of nesprin-1 in nuclear deformation in endothelial cells under static and uniaxial stretching conditions Biochem. Biophys Res Commun 424:94–99. https://doi.org/10.1016/j.bbrc.2012.06.073
Arsenovic PT et al (2016) Nesprin-2G, a component of the Nuclear LINC Complex, is subject to myosin-dependent tension. Biophys J 110:34–43. https://doi.org/10.1016/j.bpj.2015.11.014
Booth-Gauthier EA, Du V, Ghibaudo M, Rape AD, Dahl KN, Ladoux B (2013) Hutchinson-Gilford progeria syndrome alters nuclear shape and reduces cell motility in three dimensional model substrates. Integr Biol (Camb) 5:569–577. https://doi.org/10.1039/c3ib20231c
Chancellor TJ, Lee J, Thodeti CK, Lele T (2010) Actomyosin tension exerted on the nucleus through nesprin-1 connections influences endothelial cell adhesion, migration, and cyclic strain-induced reorientation. Biophys J 99:115–123. https://doi.org/10.1016/j.bpj.2010.04.011
Crisp M et al (2006) Coupling of the nucleus and cytoplasm: role of the LINC complex. J Cell Biol 172:41–53. https://doi.org/10.1083/jcb.200509124
Dahl KN, Scaffidi P, Islam MF, Yodh AG, Wilson KL, Misteli T (2006) Distinct structural and mechanical properties of the nuclear lamina in Hutchinson-Gilford progeria syndrome P natl. Acad Sci USA 103:10271–10276. https://doi.org/10.1073/pnas.0601058103
Deguchi S, Maeda K, Ohashi T, Sato M (2005) Flow-induced hardening of endothelial nucleus as an intracellular stress-bearing organelle. J Biomech 38:1751–1759. https://doi.org/10.1016/j.jbiomech.2005.06.003
Denis KB, Cabe JI, Danielsson BE, Tieu KV, Mayer CR, Conway DE (2021) The LINC complex is required for endothelial cell adhesion and adaptation to shear stress and cyclic stretch. Mol Biol Cell 32:1654–1663. https://doi.org/10.1091/mbc.E20-11-0698
Driscoll TP, Cosgrove BD, Heo SJ, Shurden ZE, Mauck RL (2015) Cytoskeletal to nuclear strain transfer regulates YAP Signaling in Mesenchymal Stem. Cells Biophys J 108:2783–2793. https://doi.org/10.1016/j.bpj.2015.05.010
Eckert J, van Loon J, Eng LM, Schmidt T (2021) Hypergravity affects cell traction forces of fibroblasts. Biophys J 120:773–780. https://doi.org/10.1016/j.bpj.2021.01.021
Engler AJ, Sen S, Sweeney HL, Discher DE (2006) Matrix elasticity directs stem cell. Lineage Specification Cell 126:677–689. https://doi.org/10.1016/j.cell.2006.06.044
Fu X et al (2024) Targeting nuclear mechanics mitigates the fibroblast invasiveness in pathological dermal scars Induced by Matrix Stiffening Adv Sci. e2308253. https://doi.org/10.1002/advs.202308253
Ghosh K et al (2007) Cell adaptation to a physiologically relevant ECM mimic with different. Viscoelastic Prop Biomaterials 28:671–679. https://doi.org/10.1016/j.biomaterials.2006.09.038
Goffin JM, Pittet P, Csucs G, Lussi JW, Meister JJ, Hinz B (2006) Focal adhesion size controls tension-dependent recruitment of alpha-smooth muscle actin to stress fibers. J Cell Biol 172:259–268. https://doi.org/10.1083/jcb.200506179
Graham DM, Burridge K (2016) Mechanotransduction and nuclear function Curr. Opin Cell Biol 40:98–105. https://doi.org/10.1016/j.ceb.2016.03.006
Graham DM et al (2018) Enucleated cells reveal differential roles of the nucleus in cell migration, polarity, and mechanotransduction. J Cell Biol 217:895–914. https://doi.org/10.1083/jcb.201706097
Guilluy C, Osborne LD, Van Landeghem L, Sharek L, Superfine R, Garcia-Mata R, Burridge K (2014) Isolated nuclei adapt to force and reveal a mechanotransduction pathway in the nucleus. Nat Cell Biol 16:376–381. https://doi.org/10.1038/ncb2927
Hale CM et al (2008) Dysfunctional connections between the nucleus and the actin and microtubule networks in laminopathic models. Biophys J 95:5462–5475. https://doi.org/10.1529/biophysj.108.139428
Ii S, Ito K, Takakusaki N, Sakamoto N (2019) Inverse estimation of 3-D traction stress field of adhered cell based on optimal control technique using. Image Intensities Mol Cell Biomech 16:49. https://doi.org/10.32604/mcb.2019.07378
Kasas S et al (2005) Superficial and deep changes of cellular mechanical properties following cytoskeleton disassembly. Cell Motil Cytoskeleton 62:124–132. https://doi.org/10.1002/cm.20086
Khatau SB et al (2009) A perinuclear actin cap regulates nuclear shape. Proc Natl Acad Sci U S A 106:19017–19022. https://doi.org/10.1073/pnas.0908686106
Kimura S et al (2020) Tissue-scale tensional homeostasis in skin regulates structure and physiological function. Commun Biol 3:637. https://doi.org/10.1038/s42003-020-01365-7
Kong HJ, Polte TR, Alsberg E, Mooney DJ (2005) FRET measurements of cell-traction forces and nano-scale clustering of adhesion ligands varied by substrate stiffness. Proc Natl Acad Sci U S A 102:4300–4305. https://doi.org/10.1073/pnas.0405873102
Ladoux B, Mege RM (2017) Mechanobiology of collective cell behaviours. Nat Rev Mol Cell Biol 18:743–757. https://doi.org/10.1038/nrm.2017.98
Li F, Li B, Wang QM, Wang JH (2008) Cell shape regulates collagen type I expression in human tendon fibroblasts. Cell Motil Cytoskeleton 65:332–341. https://doi.org/10.1002/cm.20263
Lo CM, Wang HB, Dembo M, Wang YL (2000) Cell movement is guided by the rigidity of the substrate. Biophys J 79:144–152. https://doi.org/10.1016/S0006-3495(00)76279-5
Lovett DB, Shekhar N, Nickerson JA, Roux KJ, Lele TP (2013) Modulation of nuclear shape by substrate rigidity cellular. Mol Bioeng 6:230–238. https://doi.org/10.1007/s12195-013-0270-2
Nagayama K (2021) A loss of nuclear-cytoskeletal interactions in vascular smooth muscle cell differentiation Induced by a Micro-grooved collagen substrate enabling the modeling of an. Vivo Cell Arrangement Bioeng (Basel) 8. https://doi.org/10.3390/bioengineering8090124
Petrie RJ, Gavara N, Chadwick RS, Yamada KM (2012) Nonpolarized signaling reveals two distinct modes of 3D cell migration. J Cell Biol 197:439–455. https://doi.org/10.1083/jcb.201201124
Peyton SR, Putnam AJ (2005) Extracellular matrix rigidity governs smooth muscle cell motility in a biphasic fashion. J Cell Physiol 204:198–209. https://doi.org/10.1002/jcp.20274
Sakamoto N, Ogawa M, Sadamoto K, Takeuchi M, Kataoka N (2017) Mechanical role of Nesprin-1-Mediated nucleus-actin filament binding in cyclic stretch-induced fibroblast elongation cellular. Mol Bioeng 10:327–338. https://doi.org/10.1007/s12195-017-0487-6
Samuel MS et al (2011) Actomyosin-mediated cellular tension drives increased tissue stiffness and beta-catenin activation to induce epidermal hyperplasia and tumor growth. Cancer Cell 19:776–791. https://doi.org/10.1016/j.ccr.2011.05.008
Sneddon IN (1965) The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile International. J Eng Sci 3:47–57
Solon J, Levental I, Sengupta K, Georges PC, Janmey PA (2007) Fibroblast adaptation and stiffness matching to soft elastic substrates. Biophys J 93:4453–4461. https://doi.org/10.1529/biophysj.106.101386
Stewart-Hutchinson PJ, Hale CM, Wirtz D, Hodzic D (2008) Structural requirements for the assembly of LINC complexes and their function in cellular mechanical stiffness. Exp Cell Res 314:1892–1905. https://doi.org/10.1016/j.yexcr.2008.02.022
Swift J et al (2013) Nuclear lamin-A scales with tissue stiffness and enhances matrix-. Dir Differ Sci 341:1240104. https://doi.org/10.1126/science.1240104
Ting LH et al (2012) Flow mechanotransduction regulates traction forces, intercellular forces, and adherens junctions. Am J Physiol Heart Circ Physiol 302:H2220–2229. https://doi.org/10.1152/ajpheart.00975.2011
Tschumperlin DJ, Ligresti G, Hilscher MB, Shah VH (2018) Mechanosensing and fibrosis. J Clin Invest 128:74–84. https://doi.org/10.1172/JCI93561
Verstraeten VL, Ji JY, Cummings KS, Lee RT, Lammerding J (2008) Increased mechanosensitivity and nuclear stiffness in Hutchinson-Gilford progeria cells: effects of farnesyltransferase inhibitors. Aging Cell 7:383–393. https://doi.org/10.1111/j.1474-9726.2008.00382.x
Vishavkarma R, Raghavan S, Kuyyamudi C, Majumder A, Dhawan J, Pullarkat PA (2014) Role of actin filaments in correlating nuclear shape and cell spreading. PLoS ONE 9:e107895. https://doi.org/10.1371/journal.pone.0107895
Wang X et al (2018) Mechanical stability of the cell nucleus - roles played by the cytoskeleton in nuclear deformation and strain recovery. J Cell Sci 131. https://doi.org/10.1242/jcs.209627
Zhang X et al (2007) Syne-1 and Syne-2 play crucial roles in myonuclear anchorage and motor. Neuron Innervation Dev 134:901–908. https://doi.org/10.1242/dev.02783
Acknowledgements
This study was supported in part by National Institutes of Health R35GM119617, Grants-in-Aid for Scientific Research from the MEXT of Japan (Nos. 18H03521, 18K19934, and 22K19898), a Tokyo Metropolitan Government Advanced Research Grant (R2-2), and the TMU Research Project for Emergent Future Society.
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K.I. and Y.U. performed experiments. and analyzed data. K.I., Y.U., D.E.C., and S.I. contributed analytic tools and analyzed data. D.E.C, J.N., and N.S designed research and wrote and edited the manuscript.
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Sakamoto, N., Ito, K., Ii, S. et al. A homeostatic role of nucleus-actin filament coupling in the regulation of cellular traction forces in fibroblasts. Biomech Model Mechanobiol (2024). https://doi.org/10.1007/s10237-024-01839-1
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DOI: https://doi.org/10.1007/s10237-024-01839-1