Inhibition of multiple myeloma‑derived exosomes uptake suppresses the functional response in bone marrow stromal cell

Zheng,Yongjiang; Tu,Chenggong; Zhang,Jingwen; Wang,Jinheng

doi:10.3892/ijo.2019.4685

Inhibition of multiple myeloma‑derived exosomes uptake suppresses the functional response in bone marrow stromal cell

Authors:
- Yongjiang Zheng
- Chenggong Tu
- Jingwen Zhang
- Jinheng Wang
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Affiliations: Department of Hematology, The Third Affiliated Hospital of Sun Yat‑Sen University, Guangzhou, Guangdong 510630, P.R. China, Key Laboratory of Protein Modification and Degradation, School of Basic Medical Sciences, Affiliated Cancer Hospital and Institute of Guangzhou Medical University, Guangzhou, Guangdong 511436, P.R. China
Published online on: January 11, 2019 https://doi.org/10.3892/ijo.2019.4685
Pages: 1061-1070

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Abstract

The communication between multiple myeloma (MM) cells and bone marrow stromal cells (BMSCs) serves a pivotal role in MM progression by supporting MM cell growth, proliferation and drug resistance. An exosomes‑based endogenous transport system has been determined as a novel mechanism of this communication by revealing the capacity for exchange of functional components between cells. An exosomes transfer‑mediated biological response in recipient cells is strongly determined by the detailed routes and mechanisms of exosomes internalization, which are diverse and can depend on surface molecules on the membrane of the vesicle and the recipient cell. Understanding the routes of exosomes uptake during MM cell‑BMSC communication is of great importance for the development of blocking strategies beneficial for MM treatment. In the present study, fluorescently‑labeled exosomes and pharmacological inhibitors, which are known to interfere with different internalization pathways, were used to characterize the cellular mechanisms involved in the uptake of MM cell‑derived exosomes by BMSCs. MM cell‑derived exosomes can promote BMSC viability and induce changes in multiple pro‑survival and pro‑proliferation pathways in BMSCs. As determined by flow cytometry and confocal microscopy, the uptake of MM cell‑derived exosomes proceeded primarily through endocytosis, via special caveolin‑dependent endocytosis, and partially through macropinocytosis and membrane fusion. Furthermore, treatment with endocytosis inhibitors suppressed the exosomes‑induced changes in pathways in BMSCs. Collectively, these results indicate that endocytosis is the primary route of internalization of MM cell‑derived exosomes by BMSCs and indicate that inhibition of exosomes uptake can interrupt the communication between MM cells and BMSCs and thus serve as a potential adjunctive strategy for MM treatment.

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1	Tkach M and Théry C: Communication by extracellular vesicles: Where we are and where we need to go. Cell. 164:1226–1232. 2016. View Article : Google Scholar : PubMed/NCBI
2	Camussi G, Deregibus MC, Bruno S, Grange C, Fonsato V and Tetta C: Exosome/microvesicle-mediated epigenetic reprogramming of cells. Am J Cancer Res. 1:98–110. 2011.PubMed/NCBI
3	Raposo G and Stoorvogel W: Extracellular vesicles: Exosomes, microvesicles, and friends. J Cell Biol. 200:373–383. 2013. View Article : Google Scholar : PubMed/NCBI
4	Théry C, Zitvogel L and Amigorena S: Exosomes: Composition, biogenesis and function. Nat Rev Immunol. 2:569–579. 2002. View Article : Google Scholar : PubMed/NCBI
5	Ratajczak J, Wysoczynski M, Hayek F, Janowska-Wieczorek A and Ratajczak MZ: Membrane-derived microvesicles: Important and underappreciated mediators of cell-to-cell communication. Leukemia. 20:1487–1495. 2006. View Article : Google Scholar : PubMed/NCBI
6	EL Andaloussi S, Mäger I and Breakefield XO: Extracellular vesicles: Biology and emerging therapeutic opportunities. Nat Rev Drug Discov. 12:347–357. 2013. View Article : Google Scholar : PubMed/NCBI
7	Colombo M, Raposo G and Théry C: Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annu Rev Cell Dev Biol. 30:255–289. 2014. View Article : Google Scholar : PubMed/NCBI
8	Mulcahy LA, Pink RC and Carter DR: Routes and mechanisms of extracellular vesicle uptake. J Extracell Vesicles. 3:32014. View Article : Google Scholar
9	O'Donoghue EJ and Krachler AM: Mechanisms of outer membrane vesicle entry into host cells. Cell Microbiol. 18:1508–1517. 2016. View Article : Google Scholar : PubMed/NCBI
10	Christianson HC, Svensson KJ, van Kuppevelt TH, Li JP and Belting M: Cancer cell exosomes depend on cell-surface heparan sulfate proteoglycans for their internalization and functional activity. Proc Natl Acad Sci USA. 110:17380–17385. 2013. View Article : Google Scholar : PubMed/NCBI
11	Ehrlich M, Boll W, Van Oijen A, Hariharan R, Chandran K, Nibert ML and Kirchhausen T: Endocytosis by random initiation and stabilization of clathrin-coated pits. Cell. 118:591–605. 2004. View Article : Google Scholar : PubMed/NCBI
12	Stephens L, Ellson C and Hawkins P: Roles of PI3Ks in leukocyte chemotaxis and phagocytosis. Curr Opin Cell Biol. 14:203–213. 2002. View Article : Google Scholar : PubMed/NCBI
13	Wang LH, Rothberg KG and Anderson RG: Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J Cell Biol. 123:1107–1117. 1993. View Article : Google Scholar : PubMed/NCBI
14	Swanson JA: Shaping cups into phagosomes and macropi-nosomes. Nat Rev Mol Cell Biol. 9:639–649. 2008. View Article : Google Scholar : PubMed/NCBI
15	Koivusalo M, Welch C, Hayashi H, Scott CC, Kim M, Alexander T, Touret N, Hahn KM and Grinstein S: Amiloride inhibits macropinocytosis by lowering submembranous pH and preventing Rac1 and Cdc42 signaling. J Cell Biol. 188:547–563. 2010. View Article : Google Scholar : PubMed/NCBI
16	Parolini I, Federici C, Raggi C, Lugini L, Palleschi S, De Milito A, Coscia C, Iessi E, Logozzi M, Molinari A, et al: Microenvironmental pH is a key factor for exosome traffic in tumor cells. J Biol Chem. 284:34211–34222. 2009. View Article : Google Scholar : PubMed/NCBI
17	Ha D, Yang N and Nadithe V: Exosomes as therapeutic drug carriers and delivery vehicles across biological membranes: Current perspectives and future challenges. Acta Pharm Sin B. 6:287–296. 2016. View Article : Google Scholar : PubMed/NCBI
18	Batrakova EV and Kim MS: Using exosomes, naturally-equipped nanocarriers, for drug delivery. J Control Release. 219:396–405. 2015. View Article : Google Scholar : PubMed/NCBI
19	Lemaire M, Deleu S, De Bruyne E, Van Valckenborgh E, Menu E and Vanderkerken K: The microenvironment and molecular biology of the multiple myeloma tumor. Adv Cancer Res. 110:19–42. 2011. View Article : Google Scholar : PubMed/NCBI
20	Manier S, Sacco A, Leleu X, Ghobrial IM and Roccaro AM: Bone marrow microenvironment in multiple myeloma progression. J Biomed Biotechnol. 2012:1574962012. View Article : Google Scholar : PubMed/NCBI
21	Wang J, Faict S, Maes K, De Bruyne E, Van Valckenborgh E, Schots R, Vanderkerken K and Menu E: Extracellular vesicle cross-talk in the bone marrow microenvironment: Implications in multiple myeloma. Oncotarget. 7:38927–38945. 2016.PubMed/NCBI
22	Wang J, De Veirman K, Faict S, Frassanito MA, Ribatti D, Vacca A and Menu E: Multiple myeloma exosomes establish a favourable bone marrow microenvironment with enhanced angiogenesis and immunosuppression. J Pathol. 239:162–173. 2016. View Article : Google Scholar : PubMed/NCBI
23	Wang J, De Veirman K, De Beule N, Maes K, De Bruyne E, Van Valckenborgh E, Vanderkerken K and Menu E: The bone marrow microenvironment enhances multiple myeloma progression by exosome-mediated activation of myeloid-derived suppressor cells. Oncotarget. 6:43992–44004. 2015. View Article : Google Scholar : PubMed/NCBI
24	Wang J, Hendrix A, Hernot S, Lemaire M, De Bruyne E, Van Valckenborgh E, Lahoutte T, De Wever O, Vanderkerken K and Menu E: Bone marrow stromal cell-derived exosomes as communicators in drug resistance in multiple myeloma cells. Blood. 124:555–566. 2014. View Article : Google Scholar : PubMed/NCBI
25	De Veirman K, Wang J, Xu S, Leleu X, Himpe E, Maes K, De Bruyne E, Van Valckenborgh E, Vanderkerken K, Menu E, et al: Induction of miR-146a by multiple myeloma cells in mesenchymal stromal cells stimulates their pro-tumoral activity. Cancer Lett. 377:17–24. 2016. View Article : Google Scholar : PubMed/NCBI
26	Zuhorn IS, Kalicharan R and Hoekstra D: Lipoplex-mediated transfection of mammalian cells occurs through the cholesterol-dependent clathrin-mediated pathway of endocytosis. J Biol Chem. 277:18021–18028. 2002. View Article : Google Scholar : PubMed/NCBI
27	Uhrig S, Coutelle O, Wiehe T, Perabo L, Hallek M and Büning H: Successful target cell transduction of capsid-engineered rAAV vectors requires clathrin-dependent endocytosis. Gene Ther. 19:210–218. 2012. View Article : Google Scholar
28	Kim KS, Yoon YR, Lee HJ, Yoon S, Kim SY, Shin SW, An JJ, Kim MS, Choi SY, Sun W, et al: Enhanced hypothalamic leptin signaling in mice lacking dopamine D2 receptors. J Biol Chem. 285:8905–8917. 2010. View Article : Google Scholar : PubMed/NCBI
29	Gawecka JE, Young-Robbins SS, Sulzmaier FJ, Caliva MJ, Heikkilä MM, Matter ML and Ramos JW: RSK2 protein suppresses integrin activation and fibronectin matrix assembly and promotes cell migration. J Biol Chem. 287:43424–43437. 2012. View Article : Google Scholar : PubMed/NCBI
30	MacGibeny MA, Koyuncu OO, Wirblich C, Schnell MJ and Enquist LW: Retrograde axonal transport of rabies virus is unaffected by interferon treatment but blocked by emetine locally in axons. PLoS Pathog. 14:e10071882018. View Article : Google Scholar : PubMed/NCBI
31	Feng D, Zhao WL, Ye YY, Bai XC, Liu RQ, Chang LF, Zhou Q and Sui SF: Cellular internalization of exosomes occurs through phagocytosis. Traffic. 11:675–687. 2010. View Article : Google Scholar : PubMed/NCBI
32	Umezu T, Tadokoro H, Azuma K, Yoshizawa S, Ohyashiki K and Ohyashiki JH: Exosomal miR-135b shed from hypoxic multiple myeloma cells enhances angiogenesis by targeting factor-inhibiting HIF-1. Blood. 124:3748–3757. 2014. View Article : Google Scholar : PubMed/NCBI
33	Théry C, Ostrowski M and Segura E: Membrane vesicles as conveyors of immune responses. Nat Rev Immunol. 9:581–593. 2009. View Article : Google Scholar : PubMed/NCBI
34	Raimondo F, Morosi L, Chinello C, Magni F and Pitto M: Advances in membranous vesicle and exosome proteomics improving biological understanding and biomarker discovery. Proteomics. 11:709–720. 2011. View Article : Google Scholar : PubMed/NCBI
35	Orozco AF and Lewis DE: Flow cytometric analysis of circulating microparticles in plasma. Cytometry A. 77:502–514. 2010. View Article : Google Scholar : PubMed/NCBI
36	Meissl K, Macho-Maschler S, Müller M and Strobl B: The good and the bad faces of STAT1 in solid tumours. Cytokine. 89:12–20. 2017. View Article : Google Scholar
37	Wingelhofer B, Neubauer HA, Valent P, Han X, Constantinescu SN, Gunning PT, Müller M and Moriggl R: Implications of STAT3 and STAT5 signaling on gene regulation and chromatin remodeling in hematopoietic cancer. Leukemia. 32:1713–1726. 2018. View Article : Google Scholar : PubMed/NCBI
38	Tanimura S and Takeda K: ERK signalling as a regulator of cell motility. J Biochem. 162:145–154. 2017. View Article : Google Scholar : PubMed/NCBI
39	Costa Verdera H, Gitz-Francois JJ, Schiffelers RM and Vader P: Cellular uptake of extracellular vesicles is mediated by clathrin-independent endocytosis and macropinocytosis. J Controll Release. 266:100–108. 2017. View Article : Google Scholar
40	Doherty GJ and McMahon HT: Mechanisms of endocytosis. Annu Rev Biochem. 78:857–902. 2009. View Article : Google Scholar : PubMed/NCBI
41	Rosales C and Uribe-Querol E: Phagocytosis: A fundamental process in immunity. BioMed Res Int. 2017:90428512017. View Article : Google Scholar : PubMed/NCBI
42	Kaksonen M and Roux A: Mechanisms of clathrin-mediated endocytosis. Nat Rev Mol Cell Biol. 19:313–326. 2018. View Article : Google Scholar : PubMed/NCBI
43	Rosendale M and Perrais D: Imaging in focus: Imaging the dynamics of endocytosis. Int J Biochem Cell Biol. 93:41–45. 2017. View Article : Google Scholar : PubMed/NCBI
44	Franzen CA, Simms PE, Van Huis AF, Foreman KE, Kuo PC and Gupta GN: Characterization of uptake and internalization of exosomes by bladder cancer cells. BioMed Res Int. 2014:6198292014. View Article : Google Scholar : PubMed/NCBI
45	Mayor S and Pagano RE: Pathways of clathrin-independent endocytosis. Nat Rev Mol Cell Biol. 8:603–612. 2007. View Article : Google Scholar : PubMed/NCBI
46	Taylor MJ, Lampe M and Merrifield CJ: A feedback loop between dynamin and actin recruitment during clathrin-mediated endocytosis. PLoS Biol. 10:e10013022012. View Article : Google Scholar : PubMed/NCBI
47	Marks B, Stowell MH, Vallis Y, Mills IG, Gibson A, Hopkins CR and McMahon HT: GTPase activity of dynamin and resulting conformation change are essential for endocytosis. Nature. 410:231–235. 2001. View Article : Google Scholar : PubMed/NCBI
48	Parton RG and Simons K: The multiple faces of caveolae. Nat Rev Mol Cell Biol. 8:185–194. 2007. View Article : Google Scholar : PubMed/NCBI
49	Fitzner D, Schnaars M, van Rossum D, Krishnamoorthy G, Dibaj P, Bakhti M, Regen T, Hanisch UK and Simons M: Selective transfer of exosomes from oligodendrocytes to microglia by macropinocytosis. J Cell Sci. 124:447–458. 2011. View Article : Google Scholar : PubMed/NCBI
50	Escrevente C, Keller S, Altevogt P and Costa J: Interaction and uptake of exosomes by ovarian cancer cells. BMC Cancer. 11:1082011. View Article : Google Scholar : PubMed/NCBI
51	Kerr MC and Teasdale RD: Defining macropinocytosis. Traffic. 10:364–371. 2009. View Article : Google Scholar : PubMed/NCBI