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Employing mesenchymal stem cells to support tumor-targeted delivery of extracellular vesicle (EV)-encapsulated microRNA-379

Oncogenevolume 37pages21372149 (2018) | Download Citation

Abstract

Adult Mesenchymal Stem Cells (MSCs) have a well-established tumor-homing capacity, highlighting potential as tumor-targeted delivery vehicles. MSCs secrete extracellular vesicle (EV)-encapsulated microRNAs, which play a role in intercellular communication. The aim of this study was to characterize a potential tumor suppressor microRNA, miR-379, and engineer MSCs to secrete EVs enriched with miR-379 for in vivo therapy of breast cancer. miR-379 expression was significantly reduced in lymph node metastases compared to primary tumor tissue from the same patients. A significant reduction in the rate of tumor formation and growth in vivo was observed in T47D breast cancer cells stably expressing miR-379. In more aggressive HER2-amplified HCC-1954 cells, HCC-379 and HCC-NTC tumor growth rate in vivo was similar, but increased tumor necrosis was observed in HCC-379 tumors. In response to elevated miR-379, COX-2 mRNA and protein was also significantly reduced in vitro and in vivo. MSCs were successfully engineered to secrete EVs enriched with miR-379, with the majority found to be of the appropriate size and morphology of exosomal EVs. Administration of MSC-379 or MSC-NTC cells, or EVs derived from either cell population, resulted in no adverse effects in vivo. While MSC-379 cells did not impact tumor growth, systemic administration of cell-free EVs enriched with miR-379 was demonstrated to have a therapeutic effect. The data presented support miR-379 as a potent tumor suppressor in breast cancer, mediated in part through regulation of COX-2. Exploiting the tumor-homing capacity of MSCs while engineering the cells to secrete EVs enriched with miR-379 holds exciting potential as an innovative therapy for metastatic breast cancer.

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References

  1. 1.

    Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin. 2016;66:7–30.

  2. 2.

    Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116:281–97.

  3. 3.

    Cimmino A, Calin GA, Fabbri M, Iorio MV, Ferracin M, Shimizu M, et al. miR-15 and miR-16 induce apoptosis by targeting BCL2. Proc Natl Acad Sci USA. 2005;102:13944–9.

  4. 4.

    Calin GA, Cimmino A, Fabbri M, Ferracin M, Wojcik SE, Shimizu M, et al. MiR-15a and miR-16-1 cluster functions in human leukemia. Proc Natl Acad Sci USA. 2008;105:5166–71.

  5. 5.

    Laddha SV, Nayak S, Paul D, Reddy R, Sharma C, Jha P, et al. Genome-wide analysis reveals downregulation of miR-379/miR-656 cluster in human cancers. Biol Direct. 2013;8:10.

  6. 6.

    Khan S, Brougham CL, Ryan J, Sahrudin A, O’Neill G, Wall D, et al. miR-379 regulates cyclin B1 expression and is decreased in breast cancer. PLoS ONE. 2013;8:e68753.

  7. 7.

    Chen JS, Li HS, Huang JQ, Dong SH, Huang ZJ, Yi W, et al. MicroRNA-379-5p inhibits tumor invasion and metastasis by targeting FAK/AKT signaling in hepatocellular carcinoma. Cancer Lett. 2016;375:73–83.

  8. 8.

    Li Z, Shen J, Chan MT, Wu WK. MicroRNA-379 suppresses osteosarcoma progression by targeting PDK1. J Cell Mol Med. 2017;21:315–23.

  9. 9.

    Clancy C, Khan S, Glynn CL, Holian E, Dockery P, Lalor P, et al. Screening of exosomal microRNAs from colorectal cancer cells. Cancer Biomark. 2016;17:427–35.

  10. 10.

    Chen JS, Huang JQ, Dong SH, Huang XH. [Effects of microRNA-379-5p on proliferation, migration and invasion of hepatocellular carcinoma cell line]. Zhonghua Yi Xue Za Zhi. 2016;96:1450–3.

  11. 11.

    Li K, Wang Y, Zhang A, Liu B, Jia L. miR-379 inhibits cell proliferation, invasion, and migration of vascular smooth muscle cells by targeting insulin-like factor-1. Yonsei Med J. 2017;58:234–40.

  12. 12.

    Yamamoto K, Seike M, Takeuchi S, Soeno C, Miyanaga A, Noro R, et al. MiR-379/411 cluster regulates IL-18 and contributes to drug resistance in malignant pleural mesothelioma. Oncol Rep. 2014;32:2365–72.

  13. 13.

    Gururajan M, Josson S, Chu GC, Lu CL, Lu YT, Haga CL, et al. miR-154* and miR-379 in the DLK1-DIO3 microRNA mega-cluster regulate epithelial to mesenchymal transition and bone metastasis of prostate cancer. Clin Cancer Res. 2014;20:6559–69.

  14. 14.

    Pollari S, Leivonen SK, Perala M, Fey V, Kakonen SM, Kallioniemi O. Identification of microRNAs inhibiting TGF-beta-induced IL-11 production in bone metastatic breast cancer cells. PLoS ONE. 2012;7:e37361.

  15. 15.

    Singh-Ranger G, Mokbel K. The role of cyclooxygenase-2 (COX-2) in breast cancer, and implications of COX-2 inhibition. Eur J Surg Oncol. 2002;28:729–37.

  16. 16.

    Costa C, Soares R, Reis-Filho JS, Leitao D, Amendoeira I, Schmitt FC. Cyclo-oxygenase 2 expression is associated with angiogenesis and lymph node metastasis in human breast cancer. J Clin Pathol. 2002;55:429–34.

  17. 17.

    Ristimaki A, Sivula A, Lundin J, Lundin M, Salminen T, Haglund C, et al. Prognostic significance of elevated cyclooxygenase-2 expression in breast cancer. Cancer Res. 2002;62:632–5.

  18. 18.

    Holmes MD, Chen WY, Schnitt SJ, Collins L, Colditz GA, Hankinson SE, et al. COX-2 expression predicts worse breast cancer prognosis and does not modify the association with aspirin. Breast Cancer Res Treat. 2011;130:657–62.

  19. 19.

    Spaeth E, Klopp A, Dembinski J, Andreeff M, Marini F. Inflammation and tumor microenvironments: defining the migratory itinerary of mesenchymal stem cells. Gene Ther. 2008;15:730–8.

  20. 20.

    Dwyer RM, Potter-Beirne SM, Harrington KA, Lowery AJ, Hennessy E, Murphy JM, et al. Monocyte chemotactic protein-1 secreted by primary breast tumors stimulates migration of mesenchymal stem cells. Clin Cancer Res. 2007;13:5020–7.

  21. 21.

    Ramasamy R, Lam EW, Soeiro I, Tisato V, Bonnet D, Dazzi F. Mesenchymal stem cells inhibit proliferation and apoptosis of tumor cells: impact on in vivo tumor growth. Leukemia. 2007;21:304–10.

  22. 22.

    Momin EN, Vela G, Zaidi HA, Quinones-Hinojosa A. The oncogenic potential of mesenchymal stem cells in the treatment of cancer: directions for future research. Curr Immunol Rev. 2010;6:137–48.

  23. 23.

    Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8:315–7.

  24. 24.

    Squillaro T, Peluso G, Galderisi U. Clinical trials with mesenchymal stem cells: an update. Cell Transplant. 2016;25:829–48.

  25. 25.

    Yeo RW, Lai RC, Zhang B, Tan SS, Yin Y, Teh BJ, et al. Mesenchymal stem cell: an efficient mass producer of exosomes for drug delivery. Adv Drug Deliv Rev. 2013;65:336–41.

  26. 26.

    Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 2007;9:654–9.

  27. 27.

    Costa-Silva B, Aiello NM, Ocean AJ, Singh S, Zhang H, Thakur BK, et al. Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver. Nat Cell Biol. 2015;17:816–26.

  28. 28.

    Munoz JL, Bliss SA, Greco SJ, Ramkissoon SH, Ligon KL, Rameshwar P. Delivery of functional anti-miR-9 by mesenchymal stem cell-derived exosomes to glioblastoma multiforme cells conferred chemosensitivity. Mol Ther Acids. 2013;2:e126.

  29. 29.

    Katakowski M, Buller B, Zheng X, Lu Y, Rogers T, Osobamiro O, et al. Exosomes from marrow stromal cells expressing miR-146b inhibit glioma growth. Cancer Lett. 2013;335:201–4.

  30. 30.

    Khan S, Brougham CL, Ryan J, Sahrudin A, O’Neill G, Wall D, et al. miR-379 regulates cyclin B1 expression and is decreased in breast cancer. PLoS ONE. 2013;8:e68753.

  31. 31.

    Roggan A, Friebel M, Do Rschel K, Hahn A, Mu Ller G. Optical properties of circulating human blood in the wavelength range 400-2500 nm. J Biomed Opt. 1999;4:36–46.

  32. 32.

    Xu Z, Li C, Wang LV. Photoacoustic tomography of water in phantoms and tissue. J Biomed Opt. 2010;15:036019.

  33. 33.

    Heijblom M, Piras D, Brinkhuis M, van Hespen JCG, van den Engh FM, van der Schaaf M, et al. Photoacoustic image patterns of breast carcinoma and comparisons with magnetic resonance imaging and vascular stained histopathology. Sci Rep. 2015;5:11778.

  34. 34.

    Gilligan KE, Dwyer RM. Engineering exosomes for cancer therapy. Int J Mol Sci2017;18:pii: E1122

  35. 35.

    Alvarez-Erviti L, Seow Y, Yin H, Betts C, Lakhal S, Wood MJ. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat Biotechnol. 2011;29:341–5.

  36. 36.

    Ohno SI, Takanashi M, Sudo K, Ueda S, Ishikawa A, Matsuyama N, et al. Systemically injected exosomes targeted to EGFR deliver antitumor microRNA to breast cancer cells. Mol Ther. 2013;21:185–91.

  37. 37.

    Tian Y, Li S, Song J, Ji T, Zhu M, Anderson GJ, et al. A doxorubicin delivery platform using engineered natural membrane vesicle exosomes for targeted tumor therapy. Biomaterials. 2014;35:2383–90.

  38. 38.

    Consortium E-T, Van Deun J, Mestdagh P, Agostinis P, Akay O, Anand S, et al. EV-TRACK: transparent reporting and centralizing knowledge in extracellular vesicle research. Nat Methods. 2017;14:228–32.

  39. 39.

    Chen KH, Chen CH, Wallace CG, Yuen CM, Kao GS, Chen YL, et al. Intravenous administration of xenogenic adipose-derived mesenchymal stem cells (ADMSC) and ADMSC-derived exosomes markedly reduced brain infarct volume and preserved neurological function in rat after acute ischemic stroke. Oncotarget. 2016;7:74537–56.

  40. 40.

    Kamerkar S, LeBleu VS, Sugimoto H, Yang S, Ruivo CF, Melo SA, et al. Exosomes facilitate therapeutic targeting of oncogenic KRAS in pancreatic cancer. Nature. 2017;546:498–503.

  41. 41.

    Yang T, Martin P, Fogarty B, Brown A, Schurman K, Phipps R, et al. Exosome delivered anticancer drugs across the blood-brain barrier for brain cancer therapy in Danio rerio. Pharm Res. 2015;32:2003–14.

  42. 42.

    Lai CP, Mardini O, Ericsson M, Prabhakar S, Maguire CA, Chen JW, et al. Dynamic biodistribution of extracellular vesicles in vivo using a multimodal imaging reporter. ACS Nano. 2014;8:483–94.

  43. 43.

    Hwang DW, Choi H, Jang SC, Yoo MY, Park JY, Choi NE, et al. Noninvasive imaging of radiolabeled exosome-mimetic nanovesicle using (99m)Tc-HMPAO. Sci Rep. 2015;5:15636.

  44. 44.

    Mellin-Olsen J, Staender S, Whitaker DK, Smith AF. The Helsinki Declaration on Patient Safety in Anaesthesiology. Eur J Anaesthesiol. 2010;27:592–7.

  45. 45.

    Barry F, Murphy J. Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol. 2004;36:568–84.

  46. 46.

    Martin FT, Dwyer RM, Kelly J, Khan S, Murphy JM, Curran C, et al. Potential role of mesenchymal stem cells (MSCs) in the breast tumour microenvironment: stimulation of epithelial to mesenchymal transition (EMT). Breast Cancer Res Treat. 2010;124:317–26.

  47. 47.

    Hole P, Sillence K, Hannell C, Maguire CM, Roesslein M, Suarez G, et al. Interlaboratory comparison of size measurements on nanoparticles using nanoparticle tracking analysis (NTA). J Nanopart Res. 2013;15:2101.

  48. 48.

    Maguire MT, Boult J. Building a foundation of strength. Addressing the incidence of limb loss. Rehab Manag. 2010;23:20–3.

  49. 49.

    Livak K, Schmittgen T. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25:402–8.

  50. 50.

    Davoren PA, McNeill RE, Lowery AJ, Kerin MJ, Miller N. Identification of suitable endogenous control genes for microRNA gene expression analysis in human breast cancer. BMC Mol Biol. 2008;9:76.

  51. 51.

    McNeill RE, Miller N, Kerin MJ. Evaluation and validation of candidate endogenous control genes for real-time quantitative PCR studies of breast cancer. BMC Mol Biol. 2007;8:107.

  52. 52.

    Dwyer RM, Ryan J, Havelin RJ, Morris JC, Miller BW, Liu Z, et al. Mesenchymal stem cell (MSC) mediated delivery of the sodium iodide symporter (NIS) supports radionuclide imaging and treatment of breast cancer. Stem Cells. 2011;29:1149–57.

  53. 53.

    Dwyer RM, Potter-Beirne SM, Harrington KA, Lowery AJ, Hennessy E, Murphy JM, et al. Monocyte chemotactic protein-1 (MCP-1) secreted by primary breast tumors stimulates migration of Mesenchymal Stem Cells (MSCs). Clin Cancer Res. 2007;13:5020–7.

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Acknowledgements

The authors are grateful to Catherine Curran, Emer Hennessy, Natasha Solovyova, and Dr. Georgina Shaw for technical support provided.

Funding

KPO’B and SK: Irish Cancer Society BREAST-PREDICT collaborative cancer research center CCRGAL13; KEG: Irish Research Council (IRC) Postgraduate Scholarship GOIPG/2016/978; HZ: IRC Enterprise Partnership Scheme Postdoctoral Research Fellowship EPSPD/2016/20. JRS and KSJ: Wellcome Trust biomedical vacation scholarship; ADB: Health Research Board summer scholarship SS-2015-1365. This research was also supported by Science Foundation Ireland grants 09/SRC/B1794 and 12/RI/2338, funding from the European Union’s 7th Framework Programme under grant agreement no. HEALTH-2007-B-223298 (PurStem), and the charity Breast Cancer Research.

Author information

Affiliations

  1. Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway, Ireland

    • K. P. O’Brien
    • , S. Khan
    • , K. E. Gilligan
    • , C. Glynn
    • , A. De Bhulbh
    • , J. R. Schweber
    • , K. St John
    • , M. J. Kerin
    •  & R. M. Dwyer
  2. Cardiovascular Research Centre Galway, School of Medicine, National University of Ireland Galway, Galway, Ireland

    • H. Zafar
  3. Discipline of Anatomy, School of Medicine, National University of Ireland Galway, Galway, Ireland

    • P. Lalor
    •  & P. Dockery
  4. Regenerative Medicine Institute (REMEDI), CURAM, National University of Ireland Galway, Galway, Ireland

    • C. O’Flatharta
    • , J. M. Murphy
    •  & T. O’Brien
  5. Division of Anatomic Pathology, University Hospital Galway, Galway, Ireland

    • H. Ingoldsby
  6. Tissue Optics and Microcirculation Imaging Group, School of Physics, National University of Ireland Galway, Galway, Ireland

    • M. Leahy
  7. Cancer Biology and Therapeutics Laboratory, UCD School of Biomolecular and Biomedical Science, UCD Conway Institute, University College Dublin, Dublin, Ireland

    • W. M. Gallagher

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The authors declare that they have no competing interests.

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Correspondence to R. M. Dwyer.

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https://doi.org/10.1038/s41388-017-0116-9