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Tropomyosin Tpm 2.1 loss induces glioblastoma spreading in soft brain-like environments

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Abstract

Introduction

The brain is a very soft tissue. Glioblastoma (GBM) brain tumours are highly infiltrative into the surrounding healthy brain tissue and invasion mechanisms that have been defined using rigid substrates therefore may not apply to GBM dissemination. GBMs characteristically lose expression of the high molecular weight tropomyosins, a class of actin-associating proteins and essential regulators of the actin stress fibres and focal adhesions that underpin cell migration on rigid substrates.

Methods

Here, we investigated how loss of the high molecular weight tropomyosins affects GBM on soft matrices that recapitulate the biomechanical architecture of the brain.

Results

We find that Tpm 2.1 is down-regulated in GBM grown on soft substrates. We demonstrate that Tpm 2.1 depletion by siRNA induces cell spreading and elongation in soft 3D hydrogels, irrespective of matrix composition. Tpm 1.7, a second high molecular weight tropomyosin is also down-regulated when cells are cultured on soft brain-like surfaces and we show that effects of this isoform are matrix dependent, with Tpm 1.7 inducing cell rounding in 3D collagen gels. Finally, we show that the absence of Tpm 2.1 from primary patient-derived GBMs correlates with elongated, mesenchymal invasion.

Conclusions

We propose that Tpm 2.1 down-regulation facilitates GBM colonisation of the soft brain environment. This specialisation of the GBM actin cytoskeleton organisation that is highly suited to the soft brain-like environment may provide novel therapeutic targets for arresting GBM invasion.

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References

  1. Giese A, Bjerkvig R, Berens ME, Westphal M (2003) Cost of migration: invasion of malignant gliomas and implications for treatment. J Clin Oncol 21(8):1624–1636. https://doi.org/10.1200/jco.2003.05.063

    Article  CAS  PubMed  Google Scholar 

  2. Franze K (2013) The mechanical control of nervous system development. Development 140(15):3069–3077. https://doi.org/10.1242/dev.079145

    Article  CAS  PubMed  Google Scholar 

  3. Domingues HS, Cruz A, Chan JR, Relvas JB, Rubinstein B, Pinto IM (2018) Mechanical plasticity during oligodendrocyte differentiation and myelination. Glia 66(1):5–14. https://doi.org/10.1002/glia.23206

    Article  PubMed  Google Scholar 

  4. Engler AJ, Sen S, Sweeney HL, Discher DE (2006) Matrix elasticity directs stem cell lineage specification. Cell 126(4):677–689. https://doi.org/10.1016/j.cell.2006.06.044

    Article  CAS  PubMed  Google Scholar 

  5. Jagielska A, Norman AL, Whyte G, Vliet KJ, Guck J, Franklin RJ (2012) Mechanical environment modulates biological properties of oligodendrocyte progenitor cells. Stem Cell Dev 21(16):2905–2914. https://doi.org/10.1089/scd.2012.0189

    Article  CAS  Google Scholar 

  6. Pogoda K, Janmey PA (2018) Glial tissue mechanics and mechanosensing by glial cells. Front Cell Neurosci 12:25. https://doi.org/10.3389/fncel.2018.00025

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Pogoda K, Chin L, Georges PC, Byfield FJ, Bucki R, Kim R, Weaver M, Wells RG, Marcinkiewicz C, Janmey P (2014) Compression stiffening of brain and its effect on mechanosensing by glioma cells. New J Phys 16:075002. https://doi.org/10.1088/1367-2630/16/7/075002

    Article  PubMed  PubMed Central  Google Scholar 

  8. De Clerck YA, Shimada H, Gonzalez-Gomez I, Raffel C (1994) Tumoral invasion in the central nervous system. J Neurooncol 18(2):111–121

    Article  PubMed  Google Scholar 

  9. Giese A, Westphal M (1996) Glioma invasion in the central nervous system. Neurosurgery 39(2):235–250 (discussion 250–232)

    Article  CAS  PubMed  Google Scholar 

  10. Levental KR, Yu H, Kass L, Lakins JN, Egeblad M, Erler JT, Fong SF, Csiszar K, Giaccia A, Weninger W, Yamauchi M, Gasser DL, Weaver VM (2009) Matrix crosslinking forces tumor progression by enhancing integrin signaling. Cell 139(5):891–906. https://doi.org/10.1016/j.cell.2009.10.027

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Kaufman LJ, Brangwynne CP, Kasza KE, Filippidi E, Gordon VD, Deisboeck TS, Weitz DA (2005) Glioma expansion in collagen I matrices: analyzing collagen concentration-dependent growth and motility patterns. Biophys J 89(1):635–650. https://doi.org/10.1529/biophysj.105.061994

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Paszek MJ, Zahir N, Johnson KR, Lakins JN, Rozenberg GI, Gefen A, Reinhart-King CA, Margulies SS, Dembo M, Boettiger D, Hammer DA, Weaver VM (2005) Tensional homeostasis and the malignant phenotype. Cancer Cell 8(3):241–254

    Article  CAS  PubMed  Google Scholar 

  13. Zaman MH, Trapani LM, Sieminski AL, Mackellar D, Gong H, Kamm RD, Wells A, Lauffenburger DA, Matsudaira P (2006) Migration of tumor cells in 3D matrices is governed by matrix stiffness along with cell-matrix adhesion and proteolysis. Proc Natl Acad Sci USA 103(29):10889–10894

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Soofi SS, Last JA, Liliensiek SJ, Nealey PF, Murphy CJ (2009) The elastic modulus of matrigel as determined by atomic force microscopy. J Struct Biol 167(3):216–219. https://doi.org/10.1016/j.jsb.2009.05.005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Wong SY, Ulrich TA, Deleyrolle LP, MacKay JL, Lin JM, Martuscello RT, Jundi MA, Reynolds BA, Kumar S (2015) Constitutive activation of myosin-dependent contractility sensitizes glioma tumor-initiating cells to mechanical inputs and reduces tissue invasion. Cancer Res. https://doi.org/10.1158/0008-5472.can-13-3426

    Article  PubMed  PubMed Central  Google Scholar 

  16. Ruiz-Ontanon P, Orgaz JL, Aldaz B, Elosegui-Artola A, Martino J, Berciano MT, Montero JA, Grande L, Nogueira L, Diaz-Moralli S, Esparis-Ogando A, Vazquez-Barquero A, Lafarga M, Pandiella A, Cascante M, Segura V, Martinez-Climent JA, Sanz-Moreno V, Fernandez-Luna JL (2013) Cellular plasticity confers migratory and invasive advantages to a population of glioblastoma-initiating cells that infiltrate peritumoral tissue. Stem Cell 31(6):1075–1085. https://doi.org/10.1002/stem.1349

    Article  CAS  Google Scholar 

  17. Grundy TJ, De Leon E, Griffin KR, Stringer BW, Day BW, Fabry B, Cooper-White J, O’Neill GM (2016) Differential response of patient-derived primary glioblastoma cells to environmental stiffness. Sci Rep 6:23353. https://doi.org/10.1038/srep23353

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Kim SN, Jeibmann A, Halama K, Witte HT, Walte M, Matzat T, Schillers H, Faber C, Senner V, Paulus W, Klambt C (2014) ECM stiffness regulates glial migration in Drosophila and mammalian glioma models. Development 141(16):3233–3242. https://doi.org/10.1242/dev.106039

    Article  CAS  PubMed  Google Scholar 

  19. Lee S, Kumar S (2016) Actomyosin stress fiber mechanosensing in 2D and 3D. F1000Research. https://doi.org/10.12688/f1000research.8800.1

    Article  PubMed  PubMed Central  Google Scholar 

  20. Hughes JA, Cooke-Yarborough CM, Chadwick NC, Schevzov G, Arbuckle SM, Gunning P, Weinberger RP (2003) High-molecular-weight tropomyosins localize to the contractile rings of dividing CNS cells but are absent from malignant pediatric and adult CNS tumors. Glia 42(1):25–35

    Article  PubMed  Google Scholar 

  21. Tojkander S, Gateva G, Schevzov G, Hotulainen P, Naumanen P, Martin C, Gunning PW, Lappalainen P (2011) A molecular pathway for myosin II recruitment to stress fibers. Curr Biol 21(7):539–550. https://doi.org/10.1016/j.cub.2011.03.007

    Article  CAS  PubMed  Google Scholar 

  22. Bershadsky AD, Ballestrem C, Carramusa L, Zilberman Y, Gilquin B, Khochbin S, Alexandrova AY, Verkhovsky AB, Shemesh T, Kozlov MM (2006) Assembly and mechanosensory function of focal adhesions: experiments and models. Eur J Cell Biol 85(3–4):165–173

    Article  CAS  PubMed  Google Scholar 

  23. Wolfenson H, Meacci G, Liu S, Stachowiak MR, Iskratsch T, Ghassemi S, Roca-Cusachs P, O’Shaughnessy B, Hone J, Sheetz MP (2016) Tropomyosin controls sarcomere-like contractions for rigidity sensing and suppressing growth on soft matrices. Nat Cell Biol 18(1):33–42. https://doi.org/10.1038/ncb3277

    Article  CAS  PubMed  Google Scholar 

  24. Ulrich TA, de Juan Pardo EM, Kumar S (2009) The mechanical rigidity of the extracellular matrix regulates the structure, motility, and proliferation of glioma cells. Cancer Res 69(10):4167–4174. https://doi.org/10.1158/0008-5472.can-08-4859

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Tse JR, Engler AJ (2010) Preparation of hydrogel substrates with tunable mechanical properties. Curr Protoc Cell Biol 10:10. https://doi.org/10.1002/0471143030

    Article  Google Scholar 

  26. Day BW, Stringer BW, Wilson J, Jeffree RL, Jamieson PR, Ensbey KS, Bruce ZC, Inglis P, Allan S, Winter C, Tollesson G, Campbell S, Lucas P, Findlay W, Kadrian D, Johnson D, Robertson T, Johns TG, Bartlett PF, Osborne GW, Boyd AW (2013) Glioma surgical aspirate: a viable source of tumor tissue for experimental research. Cancers 5(2):357–371. https://doi.org/10.3390/cancers5020357

    Article  PubMed  PubMed Central  Google Scholar 

  27. Mitchell CB, O’Neill GM (2017) Rac GTPase regulation of 3D invasion in neuroblastomas lacking MYCN amplification. Cell Adhes Migr 11(1):68–79. https://doi.org/10.1080/19336918.2016.1183868

    Article  CAS  Google Scholar 

  28. Yager ML, Hughes JA, Lovicu FJ, Gunning PW, Weinberger RP, O’Neill GM (2003) Functional analysis of the actin-binding protein, tropomyosin 1, in neuroblastoma. Br J Cancer 89(5):860–863

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Schevzov G, Whittaker SP, Fath T, Lin JJ, Gunning PW (2011) Tropomyosin isoforms and reagents. Bioarchitecture 1(4):135–164. https://doi.org/10.4161/bioa.1.4.17897

    Article  PubMed  PubMed Central  Google Scholar 

  30. Bradbury PM, Turner K, Mitchell C, Griffin KR, Middlemiss S, Lau L, Dagg R, Taran E, Cooper-White J, Fabry B, O’Neill GM (2017) The focal adhesion targeting (FAT) domain of p130 Crk associated substrate (p130Cas) confers mechanosensing function. J Cell Sci 130(7):1263–1273

    Article  CAS  PubMed  Google Scholar 

  31. Sanz-Moreno V, Gadea G, Ahn J, Paterson H, Marra P, Pinner S, Sahai E, Marshall CJ (2008) Rac activation and inactivation control plasticity of tumor cell movement. Cell 135(3):510–523

    Article  CAS  PubMed  Google Scholar 

  32. Lees JG, Bach CTT, Bradbury P, Paul A, Gunning PW, O’Neill GM (2011) The actin-associating protein Tm5NM1 blocks mesenchymal motility without transition to amoeboid motility. Oncogene 30(10):1241–1251

    Article  CAS  PubMed  Google Scholar 

  33. Wolf K, Mazo I, Leung H, Engelke K, von Andrian UH, Deryugina EI, Strongin AY,, Friedl P, Br€“cker EB (2003) Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis. J Cell Biol 160(2):267–277

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Gordon VD, Valentine MT, Gardel ML, Andor-Ardo D, Dennison S, Bogdanov AA, Weitz DA, Deisboeck TS (2003) Measuring the mechanical stress induced by an expanding multicellular tumor system: a case study. Exp Cell Res 289(1):58–66

    Article  CAS  PubMed  Google Scholar 

  35. O’Neill GM (2009) The coordination between actin filaments and adhesion in mesenchymal migration. Cell Adhes Migr 3(4):355–357

    Article  Google Scholar 

  36. Bargon SD, Gunning PW, O’Neill GM (2005) The Cas family docking protein, HEF1, promotes the formation of neurite-like membrane extensions. Biochim Biophys Acta 1746(2):143–154

    Article  CAS  PubMed  Google Scholar 

  37. Beningo KA, Dembo M, Kaverina I, Small JV, Wang YL (2001) Nascent focal adhesions are responsible for the generation of strong propulsive forces in migrating fibroblasts. J Cell Biol 153(4):881–888

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Stehn JR, Haass NK, Bonello T, Desouza M, Kottyan G, Treutlein H, Zeng J, Nascimento PR, Sequeira VB, Butler TL, Allanson M, Fath T, Hill TA, McCluskey A, Schevzov G, Palmer SJ, Hardeman EC, Winlaw D, Reeve VE, Dixon I, Weninger W, Cripe TP, Gunning PW (2013) A novel class of anticancer compounds targets the actin cytoskeleton in tumor cells. Cancer Res 73(16):5169–5182. https://doi.org/10.1158/0008-5472.can-12-4501

    Article  CAS  PubMed  Google Scholar 

  39. Currier MA, Stehn JR, Swain A, Chen D, Hook J, Eiffe E, Heaton A, Brown D, Nartker BA, Eaves DW, Kloss N, Treutlein H, Zeng J, Alieva IB, Dugina VB, Hardeman EC, Gunning PW, Cripe TP (2017) Identification of cancer-targeted tropomyosin inhibitors and their synergy with microtubule drugs. Mol Cancer Ther 16(8):1555–1565. https://doi.org/10.1158/1535-7163.mct-16-0873

    Article  CAS  PubMed  Google Scholar 

  40. Brayford S, Bryce NS, Schevzov G, Haynes EM, Bear JE, Hardeman EC, Gunning PW (2016) Tropomyosin promotes lamellipodial persistence by collaborating with Arp2/3 at the leading edge. Curr Biol 26(10):1312–1318. https://doi.org/10.1016/j.cub.2016.03.028

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank Janine Woehlk, Dr Kylie Turner, Kaitlyn Griffin and Dr Peta Bradbury for technical assistance.

Funding

CM, BS, BD and GO are members of the Brain Cancer Discovery Collaborative, supported by the Cure Brain Cancer Foundation and the work was supported by a University of Sydney Strategic Priority Areas for Collaboration (SPARC) cancer grant to MB and GON.

Author information

Author notes

  1. Camilla B. Mitchell

    Present address: Division of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia

Authors and Affiliations

  1. Children’s Cancer Research Unit, Kids Research Institute, The Children’s Hospital at Westmead, Locked Bag 4001, Westmead, NSW, 2145, Australia

    Camilla B. Mitchell, Bronte Black, Faith Sun & Geraldine M. O’Neill

  2. Faculty of Pharmacy, The University of Sydney, Sydney, NSW, 2006, Australia

    Wojciech Chrzanowski

  3. The University of Sydney Nano Institute, Sydney, NSW, 2006, Australia

    Wojciech Chrzanowski

  4. Tissue Engineering and Microfluidics Group, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, 4072, Australia

    Justin Cooper-White

  5. School of Chemical Engineering, The University of Queensland, Brisbane, 4072, Australia

    Justin Cooper-White

  6. CSIRO, Manufacturing Flagship, Biomedical Manufacturing Program, Clayton, VIC, Australia

    Justin Cooper-White & Benois Maisonneuve

  7. QIMR Berghofer Medical Research Institute, Herston, QLD, Australia

    Brett Stringer & Bryan Day

  8. EMBL Australia Single Molecule Science Node, School of Medical Sciences, The University of New South Wales, Sydney, Australia

    Maté Biro

  9. Centenary Institute, Sydney Medical School, The University of Sydney, Sydney, Australia

    Maté Biro

  10. Discipline of Childhood and Adolescent Health, The University of Sydney, Sydney, NSW, Australia

    Geraldine M. O’Neill

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  1. Camilla B. Mitchell
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Correspondence to Geraldine M. O’Neill.

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Mitchell, C.B., Black, B., Sun, F. et al. Tropomyosin Tpm 2.1 loss induces glioblastoma spreading in soft brain-like environments. J Neurooncol 141, 303–313 (2019). https://doi.org/10.1007/s11060-018-03049-z

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  • DOI: https://doi.org/10.1007/s11060-018-03049-z

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