Top

Published in:

01-02-2019 | Preclinical study

Plasma exosomes stimulate breast cancer metastasis through surface interactions and activation of FAK signaling

Authors: Tatiana Shtam, Stanislav Naryzhny, Roman Samsonov, David Karasik, Igor Mizgirev, Artur Kopylov, Elena Petrenko, Yana Zabrodskaya, Roman Kamyshinsky, Daniil Nikitin, Maxim Sorokin, Anton Buzdin, Hava Gil-Henn, Anastasia Malek

Published in: Breast Cancer Research and Treatment | Issue 1/2019

Abstract

Purpose

The interaction between malignant cells and surrounding healthy tissues is a critical factor in the metastatic progression of breast cancer (BC). Extracellular vesicles, especially exosomes, are known to be involved in inter-cellular communication during cancer progression. In the study presented herein, we aimed to evaluate the role of circulating plasma exosomes in the metastatic dissemination of BC and to investigate the underlying molecular mechanisms of this phenomenon.

Methods

Exosomes isolated from plasma of healthy female donors were applied in various concentrations into the medium of MDA-MB-231 and MCF-7 cell lines. Motility and invasive properties of BC cells were examined by random migration and Transwell invasion assays, and the effect of plasma exosomes on the metastatic dissemination of BC cells was demonstrated in an in vivo zebrafish model. To reveal the molecular mechanism of interaction between plasma exosomes and BC cells, a comparison between un-treated and enzymatically modified exosomes was performed, followed by mass spectrometry, gene ontology, and pathway analysis.

Results

Plasma exosomes stimulated the adhesive properties, two-dimensional random migration, and transwell invasion of BC cells in vitro as well as their in vivo metastatic dissemination in a dose-dependent manner. This stimulatory effect was mediated by interactions of surface exosome proteins with BC cells and consequent activation of focal adhesion kinase (FAK) signaling in the tumor cells.

Conclusions

Plasma exosomes have a potency to stimulate the metastasis-promoting properties of BC cells. This pro-metastatic property of normal plasma exosomes may have impact on the course of the disease and on its prognosis.

Available only for authorised users

Menard JA, Cerezo-Magana M, Belting M (2018) Functional role of extracellular vesicles and lipoproteins in the tumour microenvironment. Philos Trans R Soc Lond Ser 373 (1737). https://doi.org/10.1098/rstb.2016.0480

Green TM, Alpaugh ML, Barsky SH, Rappa G, Lorico A (2015) Breast cancer-derived extracellular vesicles: characterization and contribution to the metastatic phenotype. BioMed Res Int 2015:634865. https://doi.org/10.1155/2015/634865 CrossRefPubMedPubMedCentral

Harris DA, Patel SH, Gucek M, Hendrix A, Westbroek W, Taraska JW (2015) Exosomes released from breast cancer carcinomas stimulate cell movement. PLoS ONE 10(3):e0117495. https://doi.org/10.1371/journal.pone.0117495 CrossRefPubMedPubMedCentral

Luga V, Wrana JL (2013) Tumor-stroma interaction: revealing fibroblast-secreted exosomes as potent regulators of Wnt-planar cell polarity signaling in cancer metastasis. Cancer Res 73(23):6843–6847. https://doi.org/10.1158/0008-5472.CAN-13-1791 CrossRefPubMed

Gernapudi R, Yao Y, Zhang Y, Wolfson B, Roy S, Duru N, Eades G, Yang P, Zhou Q (2015) Targeting exosomes from preadipocytes inhibits preadipocyte to cancer stem cell signaling in early-stage breast cancer. Breast Cancer Res Treat 150(3):685–695. https://doi.org/10.1007/s10549-015-3326-2 CrossRefPubMedPubMedCentral

Lin R, Wang S, Zhao RC (2013) Exosomes from human adipose-derived mesenchymal stem cells promote migration through Wnt signaling pathway in a breast cancer cell model. Mol Cell Biochem 383(1–2):13–20. https://doi.org/10.1007/s11010-013-1746-z CrossRefPubMed

Jang JY, Lee JK, Jeon YK, Kim CW (2013) Exosome derived from epigallocatechin gallate treated breast cancer cells suppresses tumor growth by inhibiting tumor-associated macrophage infiltration and M2 polarization. BMC Cancer 13:421. https://doi.org/10.1186/1471-2407-13-421 CrossRefPubMedPubMedCentral

Chow A, Zhou W, Liu L, Fong MY, Champer J, Van Haute D, Chin AR, Ren X, Gugiu BG, Meng Z, Huang W, Ngo V, Kortylewski M, Wang SE (2014) Macrophage immunomodulation by breast cancer-derived exosomes requires Toll-like receptor 2-mediated activation of NF-kappaB. Sci Rep 4:5750. https://doi.org/10.1038/srep05750 CrossRefPubMedPubMedCentral

Whiteside TL (2017) Exosomes in cancer: another mechanism of tumor-induced immune suppression. Adv Exp Med Biol 1036:81–89. https://doi.org/10.1007/978-3-319-67577-0_6 CrossRefPubMed

10.

Rana S, Malinowska K, Zoller M (2013) Exosomal tumor microRNA modulates premetastatic organ cells. Neoplasia 15(3):281–295CrossRefPubMedPubMedCentral

11.

Costa-Silva B, Aiello NM, Ocean AJ, Singh S, Zhang H, Thakur BK, Becker A, Hoshino A, Mark MT, Molina H, Xiang J, Zhang T, Theilen TM, Garcia-Santos G, Williams C, Ararso Y, Huang Y, Rodrigues G, Shen TL, Labori KJ, Lothe IM, Kure EH, Hernandez J, Doussot A, Ebbesen SH, Grandgenett PM, Hollingsworth MA, Jain M, Mallya K, Batra SK, Jarnagin WR, Schwartz RE, Matei I, Peinado H, Stanger BZ, Bromberg J, Lyden D (2015) Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver. Nat Cell Biol 17(6):816–826. https://doi.org/10.1038/ncb3169 CrossRefPubMedPubMedCentral

12.

Hoshino A, Costa-Silva B, Shen TL, Rodrigues G, Hashimoto A, Tesic Mark M, Molina H, Kohsaka S, Di Giannatale A, Ceder S, Singh S, Williams C, Soplop N, Uryu K, Pharmer L, King T, Bojmar L, Davies AE, Ararso Y, Zhang T, Zhang H, Hernandez J, Weiss JM, Dumont-Cole VD, Kramer K, Wexler LH, Narendran A, Schwartz GK, Healey JH, Sandstrom P, Labori KJ, Kure EH, Grandgenett PM, Hollingsworth MA, de Sousa M, Kaur S, Jain M, Mallya K, Batra SK, Jarnagin WR, Brady MS, Fodstad O, Muller V, Pantel K, Minn AJ, Bissell MJ, Garcia BA, Kang Y, Rajasekhar VK, Ghajar CM, Matei I, Peinado H, Bromberg J, Lyden D (2015) Tumour exosome integrins determine organotropic metastasis. Nature 527(7578):329–335. https://doi.org/10.1038/nature15756 CrossRefPubMedPubMedCentral

13.

Ochieng J, Pratap S, Khatua AK, Sakwe AM (2009) Anchorage-independent growth of breast carcinoma cells is mediated by serum exosomes. Exp Cell Res 315(11):1875–1888. https://doi.org/10.1016/j.yexcr.2009.03.010 CrossRefPubMedPubMedCentral

14.

Almendros I, Khalyfa A, Trzepizur W, Gileles-Hillel A, Huang L, Akbarpour M, Andrade J, Farre R, Gozal D (2016) Tumor cell malignant properties are enhanced by circulating exosomes in sleep apnea. Chest 150(5):1030–1041. https://doi.org/10.1016/j.chest.2016.08.1438 CrossRefPubMed

15.

Khalyfa A, Almendros I, Gileles-Hillel A, Akbarpour M, Trzepizur W, Mokhlesi B, Huang L, Andrade J, Farre R, Gozal D (2016) Circulating exosomes potentiate tumor malignant properties in a mouse model of chronic sleep fragmentation. Oncotarget 7(34):54676–54690. https://doi.org/10.18632/oncotarget.10578 CrossRefPubMedPubMedCentral

16.

Genna A, Lapetina S, Lukic N, Twafra S, Meirson T, Sharma VP, Condeelis JS, Gil-Henn H (2018) Pyk2 and FAK differentially regulate invadopodia formation and function in breast cancer cells. J Cell Biol 217(1):375–395. https://doi.org/10.1083/jcb.201702184 CrossRefPubMedPubMedCentral

17.

Thery C, Amigorena S, Raposo G, Clayton A (2006) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol 3: 22. https://doi.org/10.1002/0471143030.cb0322s30 CrossRefPubMed

18.

Samsonov R, Burdakov V, Shtam T, Radzhabova Z, Vasilyev D, Tsyrlina E, Titov S, Ivanov M, Berstein L, Filatov M, Kolesnikov N, Gil-Henn H, Malek A (2016) Plasma exosomal miR-21 and miR-181a differentiates follicular from papillary thyroid cancer. Tumour Biol 37(9):12011–12021. https://doi.org/10.1007/s13277-016-5065-3 CrossRefPubMed

19.

Shtam T, Kovalev R, Varfolomeeva E, Makarov E, Kil Y, Filatov M (2013) Exosomes are natural carriers of exogenous siRNA to human cells in vitro. Cell Commun Signal 11:88. https://doi.org/10.1186/1478-811X-11-88 CrossRefPubMedPubMedCentral

20.

Alamri Y, Vogel R, MacAskill M, Anderson T (2016) Plasma exosome concentration may correlate with cognitive impairment in Parkinson’s disease. Alzheimer’s Dement 4:107–108. https://doi.org/10.1016/j.dadm.2016.08.002 CrossRef

21.

Shtam T, Samsonov R, Kamyshinsky R, Pantina R, Verlov N, Vasiliev A, Konevega A, Malek A (2017) Exosomes: Some approaches to cancer diagnosis and therapy. AIP Conference Proceedings 1882 (1), 020066

22.

Shtam T, Samsonov R, Volnitskiy A, Kamyshinsky R, Verlov N, Kniazeva M, Korobkina E, Orehov A, Vasiliev A, Konevega A, Malek A (2018) Isolation of extracellular micro-vesicles from cell culture medium: comparative evaluation of methods. Biomeditsinskaia Khimiia 64(1):23–30. https://doi.org/10.18097/PBMC20186401023 CrossRefPubMed

23.

Egorov VV, Lebedev DV, Shaldzhyan AA, Sirotkin AK, Gorshkov AN, Mirgorodskaya OA, Grudinina NA, Vasin AV, Shavlovsky MM (2014) A conservative mutant of a proteolytic fragment produced during fibril formation enhances fibrillogenesis. Prion 8(5):369–373. https://doi.org/10.4161/19336896.2014.983745 CrossRefPubMedPubMedCentral

24.

Nečas D, Klapetek P (2012) Gwyddion: an open-source software for SPM data analysis. Cent Eur J Phys 10(1):181–188

25.

Wisniewski JR, Zougman A, Nagaraj N, Mann M (2009) Universal sample preparation method for proteome analysis. Nat Methods 6(5):359–362. https://doi.org/10.1038/nmeth.1322 CrossRefPubMed

26.

Naryzhny S, Zgoda V, Kopylov A, Petrenko E, Kleist O, Archakov A (2017) Variety and dynamics of proteoforms in the human proteome: aspects of markers for hepatocellular carcinoma. Proteomes 5 (4). https://doi.org/10.3390/proteomes5040033

27.

Naryzhny S, Maynskova M, Zgoda V, Archakov А (2017) Zipf’s law in proteomics. J Proteom Bioinform 10:79–84CrossRef

28.

Ishihama Y, Oda Y, Tabata T, Sato T, Nagasu T, Rappsilber J, Mann M (2005) Exponentially modified protein abundance index (emPAI) for estimation of absolute protein amount in proteomics by the number of sequenced peptides per protein. Mole Cell Proteom 4(9):1265–1272. https://doi.org/10.1074/mcp.M500061-MCP200 CrossRef

29.

Ishihama Y, Schmidt T, Rappsilber J, Mann M, Hartl FU, Kerner MJ, Frishman D (2008) Protein abundance profiling of the Escherichia coli cytosol. BMC Genom 9:102. https://doi.org/10.1186/1471-2164-9-102 CrossRef

30.

Huang da W, Sherman BT, Lempicki RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4(1):44–57. https://doi.org/10.1038/nprot.2008.211 CrossRefPubMed

31.

Huang da W, Sherman BT, Lempicki RA (2009) Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res 37(1):1–13. https://doi.org/10.1093/nar/gkn923 CrossRefPubMed

32.

Nakaya A, Katayama T, Itoh M, Hiranuka K, Kawashima S, Moriya Y, Okuda S, Tanaka M, Tokimatsu T, Yamanishi Y, Yoshizawa AC, Kanehisa M, Goto S (2013) KEGG OC: a large-scale automatic construction of taxonomy-based ortholog clusters. Nucleic Acids Res 41(Database issue):D353–D357. https://doi.org/10.1093/nar/gks1239 CrossRefPubMed

33.

Schaefer CF, Anthony K, Krupa S, Buchoff J, Day M, Hannay T, Buetow KH (2009) PID: the pathway interaction database. Nucleic Acids Res 37: D674-D679. https://doi.org/10.1093/nar/gkn653 CrossRef

34.

Croft D, Mundo AF, Haw R, Milacic M, Weiser J, Wu G, Caudy M, Garapati P, Gillespie M, Kamdar MR, Jassal B, Jupe S, Matthews L, May B, Palatnik S, Rothfels K, Shamovsky V, Song H, Williams M, Birney E, Hermjakob H, Stein L, D’Eustachio P (2014) The reactome pathway knowledgebase. Nucleic Acids Res 42:D472–D477. https://doi.org/10.1093/nar/gkt1102 CrossRefPubMed

35.

Malek A, Catapano CV, Czubayko F, Aigner A (2010) A sensitive polymerase chain reaction-based method for detection and quantification of metastasis in human xenograft mouse models. Clin Exp Metastasis 27(4):261–271. https://doi.org/10.1007/s10585-010-9324-1 CrossRefPubMed

36.

The Transwell Migration Assay (2017) J Vis Exp

37.

White RM, Sessa A, Burke C, Bowman T, LeBlanc J, Ceol C, Bourque C, Dovey M, Goessling W, Burns CE, Zon LI (2008) Transparent adult zebrafish as a tool for in vivo transplantation analysis. Cell Stem Cell 2(2):183–189. https://doi.org/10.1016/j.stem.2007.11.002 CrossRefPubMedPubMedCentral

38.

Lotvall J, Hill AF, Hochberg F, Buzas EI, Di Vizio D, Gardiner C, Gho YS, Kurochkin IV, Mathivanan S, Quesenberry P, Sahoo S, Tahara H, Wauben MH, Witwer KW, Thery C (2014) Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. J Extracell Vesicles 3:26913. https://doi.org/10.3402/jev.v3.26913 CrossRefPubMed

39.

Shtam T, Naryzhny S, Kopylov A, Petrenko E, Samsonov R, Kamyshinsky R, Zabrodskaya Y, Nikitin D, Sorokin M, Buzdin A, Malek A (2018) Functional properties of circulating exosomes mediated by surface-attached plasma proteins. J Hematol 7(4):149–153CrossRefPubMedPubMedCentral

40.

Purushothaman A, Bandari SK, Liu J, Mobley JA, Brown EE, Sanderson RD (2016) Fibronectin on the surface of myeloma cell-derived exosomes mediates exosome-cell interactions. J Biol Chem 291(4):1652–1663. https://doi.org/10.1074/jbc.M115.686295 CrossRefPubMed

41.

Sung BH, Ketova T, Hoshino D, Zijlstra A, Weaver AM (2015) Directional cell movement through tissues is controlled by exosome secretion. Nat Commun 6:7164. https://doi.org/10.1038/ncomms8164 CrossRefPubMed

42.

Luga V, Zhang L, Viloria-Petit AM, Ogunjimi AA, Inanlou MR, Chiu E, Buchanan M, Hosein AN, Basik M, Wrana JL (2012) Exosomes mediate stromal mobilization of autocrine Wnt-PCP signaling in breast cancer cell migration. Cell 151(7):1542–1556. https://doi.org/10.1016/j.cell.2012.11.024 CrossRefPubMed

43.

Lakkaraju A, Rodriguez-Boulan E (2007) Cell biology: caught in the traffic. Nature 448(7151):266–267. https://doi.org/10.1038/448266a CrossRefPubMed

44.

Mu W, Rana S, Zoller M (2013) Host matrix modulation by tumor exosomes promotes motility and invasiveness. Neoplasia 15(8):875–887CrossRefPubMedPubMedCentral

45.

Koliha N, Heider U, Ozimkowski T, Wiemann M, Bosio A, Wild S (2016) Melanoma affects the composition of blood cell-derived extracellular vesicles. Front Immunol 7:282. https://doi.org/10.3389/fimmu.2016.00282 CrossRefPubMedPubMedCentral

46.

Sung BH, Weaver AM (2017) Exosome secretion promotes chemotaxis of cancer cells. Cell Adhesi Migr 11(2):187–195. https://doi.org/10.1080/19336918.2016.1273307 CrossRef

47.

Gan L, Meng J, Xu M, Liu M, Qi Y, Tan C, Wang Y, Zhang P, Weng W, Sheng W, Huang M, Wang Z (2017) Extracellular matrix protein 1 promotes cell metastasis and glucose metabolism by inducing integrin beta4/FAK/SOX2/HIF-1alpha signaling pathway in gastric cancer. Oncogene. https://doi.org/10.1038/onc.2017.363 CrossRefPubMedPubMedCentral

48.

Naik MU, Naik TU, Summer R, Naik UP (2017) Binding of CIB1 to the αIIb tail of αIIbβ3 is required for FAK recruitment and activation in platelets. PloS one 12(5):e0176602. https://doi.org/10.1371/journal.pone.0176602 CrossRefPubMedPubMedCentral

49.

Keasey MP, Jia C, Pimentel LF, Sante RR, Lovins C, Hagg T (2018) Blood vitronectin is a major activator of LIF and IL-6 in the brain through integrin-FAK and uPAR signaling. J Cell Sci. https://doi.org/10.1242/jcs.202580 CrossRefPubMedPubMedCentral

50.

Li CL, Yang D, Cao X, Wang F, Hong DY, Wang J, Shen XC, Chen Y (2017) Fibronectin induces epithelial-mesenchymal transition in human breast cancer MCF-7 cells via activation of calpain. Oncol Lett 13(5):3889–3895. https://doi.org/10.3892/ol.2017.5896 CrossRefPubMedPubMedCentral

51.

Hu C, Wen J, Gong L, Chen X, Wang J, Hu F, Zhou Q, Liang J, Wei L, Shen Y, Zhang W (2017) Thrombospondin-1 promotes cell migration, invasion and lung metastasis of osteosarcoma through FAK dependent pathway. Oncotarget 8(44):75881–75892. https://doi.org/10.18632/oncotarget.17427 CrossRefPubMedPubMedCentral

52.

Lawson C, Lim ST, Uryu S, Chen XL, Calderwood DA, Schlaepfer DD (2012) FAK promotes recruitment of talin to nascent adhesions to control cell motility. J Cell Biol 196(2):223–232. https://doi.org/10.1083/jcb.201108078 CrossRefPubMedPubMedCentral

Title: Plasma exosomes stimulate breast cancer metastasis through surface interactions and activation of FAK signaling
Authors: Tatiana Shtam
Stanislav Naryzhny
Roman Samsonov
David Karasik
Igor Mizgirev
Artur Kopylov
Elena Petrenko
Yana Zabrodskaya
Roman Kamyshinsky
Daniil Nikitin
Maxim Sorokin
Anton Buzdin
Hava Gil-Henn
Anastasia Malek
Publication date: 01-02-2019
Publisher: Springer US
Published in: Breast Cancer Research and Treatment / Issue 1/2019
Print ISSN: 0167-6806
Electronic ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-018-5043-0

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2019

A real-world study of cardiac events in > 3700 patients with HER2-positive early breast cancer treated with trastuzumab: final analysis of the OHERA study

Prognostic impact and possible pathogenesis of lymph node metastasis in ductal carcinoma in situ of the breast

Trends in use of bilateral prophylactic mastectomy vs high-risk surveillance in unaffected carriers of inherited breast cancer syndromes in the Inherited Cancer Registry (ICARE)

Time-varying risks of second events following a DCIS diagnosis in the population-based Vermont DCIS cohort

Survivorship issues in older breast cancer survivors

Connexin 43 is an independent predictor of patient outcome in breast cancer patients

Keynote webinar | Spotlight on antibody–drug conjugates in cancer