Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

Online
ISSN
1437-4331
See all formats and pricing
More options …
Volume 57, Issue 7

Issues

Extracellular vesicle-associated miRNAs in ovarian cancer – design of an integrated NGS-based workflow for the identification of blood-based biomarkers for platinum-resistance

Jan Dominik Kuhlmann
  • Corresponding author
  • Department of Gynecology and Obstetrics, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany
  • National Center for Tumor Diseases (NCT), Partner Site Dresden, Dresden, Germany
  • German Cancer Consortium (DKTK), Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Issam Chebouti
  • German Cancer Consortium (DKTK), Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany
  • Department of Gynecology and Obstetrics, University Hospital Essen, Essen, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Rainer Kimmig
  • German Cancer Consortium (DKTK), Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany
  • Department of Gynecology and Obstetrics, University Hospital Essen, Essen, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Paul Buderath
  • German Cancer Consortium (DKTK), Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany
  • Department of Gynecology and Obstetrics, University Hospital Essen, Essen, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Michael Reuter
  • Department of Diagnostics, Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Sven-Holger Puppel
  • Department of Diagnostics, Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Pauline Wimberger
  • Department of Gynecology and Obstetrics, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany
  • National Center for Tumor Diseases (NCT), Partner Site Dresden, Dresden, Germany
  • German Cancer Consortium (DKTK), Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Sabine Kasimir-Bauer
  • German Cancer Consortium (DKTK), Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany
  • Department of Gynecology and Obstetrics, University Hospital Essen, Essen, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2018-11-13 | DOI: https://doi.org/10.1515/cclm-2018-1048

Abstract

Background

Extracellular vesicle (EV)-associated microRNAs (miRNAs) have been suggested as promising biomarkers for blood-based cancer diagnosis. However, one of the major limitations for the use of EVs with diagnostic purpose is the lack of standardized EV-profiling techniques. In this regard, the objective of our study was to design an integrated next-generation sequencing (NGS)-based workflow for analyzing the signature of EV-associated miRNA in the plasma of platinum-resistant ovarian cancer patients.

Methods

For EV-extraction, different enrichment methods were compared (ExoQuick vs. exoRNeasy). NGS was performed with the Illumina platform.

Results

We established an integrated NGS-based workflow, including EV-enrichment with the ExoQuick system, which resulted in an optimal RNA-yield and consistent small RNA libraries. We applied this workflow in a pilot cohort of clinically documented platinum-sensitive (n=15) vs. platinum-resistant (n=15) ovarian cancer patients, resulting in a panel of mature EV-associated miRNAs (including ovarian cancer associated miR-181a, miR-1908, miR-21, miR-486 and miR-223), which were differentially abundant in the plasma of platinum-resistant patients.

Conclusions

This is the first study, analyzing the profile of EV-associated miRNAs in platinum-resistant ovarian cancer patients. We provide rationale to further validate these miRNA candidates in an independent set of patients, in order to characterize their biomarker potential as predictors for platinum-resistance.

This article offers supplementary material which is provided at the end of the article.

Keywords: extracellular vesicles; microRNAs; next-generation-sequencing; ovarian cancer; platinum-resistance

References

  • 1.

    Goodman MT, Howe HL, Tung KH, Hotes J, Miller BA, Coughlin SS, et al. Incidence of ovarian cancer by race and ethnicity in the United States, 1992–1997. Cancer 2003;97:2676–85.PubMedCrossrefGoogle Scholar

  • 2.

    du Bois A, Quinn M, Thigpen T, Vermorken J, Avall-Lundqvist E, Bookman M, et al. 2004 consensus statements on the management of ovarian cancer: final document of the 3rd International Gynecologic Cancer Intergroup Ovarian Cancer Consensus Conference (GCIG OCCC 2004). Ann Oncol 2005;16(Suppl. 8): viii7–12.PubMedGoogle Scholar

  • 3.

    du Bois A, Reuss A, Pujade-Lauraine E, Harter P, Ray-Coquard I, Pfisterer J. Role of surgical outcome as prognostic factor in advanced epithelial ovarian cancer: a combined exploratory analysis of 3 prospectively randomized phase 3 multicenter trials: by the Arbeitsgemeinschaft Gynaekologische Onkologie Studiengruppe Ovarialkarzinom (AGO-OVAR) and the Groupe d’investigateurs Nationaux Pour les Etudes des Cancers de l’Ovaire (GINECO). Cancer 2009;115:1234–44.PubMedWeb of ScienceGoogle Scholar

  • 4.

    Wimberger P, Lehmann N, Kimmig R, Burges A, Meier W, Du Bois A, et al. Prognostic factors for complete debulking in advanced ovarian cancer and its impact on survival. An exploratory analysis of a prospectively randomized phase III study of the Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study Group (AGO-OVAR). Gynecol Oncol 2007;106:69–74.Web of ScienceGoogle Scholar

  • 5.

    Wimberger P, Wehling M, Lehmann N, Kimmig R, Schmalfeldt B, Burges A, et al. Influence of residual tumor on outcome in ovarian cancer patients with FIGO stage IV disease: an exploratory analysis of the AGO-OVAR (Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study Group). Ann Surg Oncol 2010;17:1642–8.CrossrefGoogle Scholar

  • 6.

    Martin LP, Schilder RJ. Management of recurrent ovarian carcinoma: current status and future directions. Semin Oncol 2009;36:112–25.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 7.

    Burger RA, Brady MF, Bookman MA, Fleming GF, Monk BJ, Huang H, et al. Incorporation of bevacizumab in the primary treatment of ovarian cancer. N Engl J Med 2011;365:2473–83.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 8.

    Ledermann J, Harter P, Gourley C, Friedlander M, Vergote I, Rustin G, et al. Olaparib maintenance therapy in patients with platinum-sensitive relapsed serous ovarian cancer: a preplanned retrospective analysis of outcomes by BRCA status in a randomised phase 2 trial. Lancet Oncol 2014;15:852–61.Web of ScienceGoogle Scholar

  • 9.

    Bookman MA. Extending the platinum-free interval in recurrent ovarian cancer: the role of topotecan in second-line chemotherapy. Oncologist 1999;4:87–94.PubMedGoogle Scholar

  • 10.

    Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL, et al. Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci USA 2008;105:10513–8.CrossrefGoogle Scholar

  • 11.

    Kuhlmann JD, Baraniskin A, Hahn SA, Mosel F, Bredemeier M, Wimberger P, et al. Circulating U2 small nuclear RNA fragments as a novel diagnostic tool for patients with epithelial ovarian cancer. Clin Chem 2014;60:206–13.CrossrefWeb of SciencePubMedGoogle Scholar

  • 12.

    Ma R, Jiang T, Kang X. Circulating microRNAs in cancer: origin, function and application. J Exp Clin Cancer Res 2012;31:38.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 13.

    Wang J, Chen J, Chang P, LeBlanc A, Li D, Abbruzzesse JL, et al. MicroRNAs in plasma of pancreatic ductal adenocarcinoma patients as novel blood-based biomarkers of disease. Cancer Prev Res (Phila) 2009;2:807–13.PubMedCrossrefGoogle Scholar

  • 14.

    Arroyo JD, Chevillet JR, Kroh EM, Ruf IK, Pritchard CC, Gibson DF, et al. Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma. Proc Natl Acad Sci USA 2011;108:5003–8.CrossrefGoogle Scholar

  • 15.

    Schwarzenbach H, Nishida N, Calin GA, Pantel K. Clinical relevance of circulating cell-free microRNAs in cancer. Nat Rev Clin Oncol 2014;11:145–56.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 16.

    Chen X, Liang H, Zhang J, Zen K, Zhang CY. Secreted microRNAs: a new form of intercellular communication. Trends Cell Biol 2012;22:125–32.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 17.

    Singh R, Pochampally R, Watabe K, Lu Z, Mo YY. Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer. Mol Cancer 2014;13:256.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 18.

    Cheng L, Wu S, Zhang K, Qing Y, Xu T. A comprehensive overview of exosomes in ovarian cancer: emerging biomarkers and therapeutic strategies. J Ovarian Res 2017;10:73.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 19.

    Crow J, Atay S, Banskota S, Artale B, Schmitt S, Godwin AK. Exosomes as mediators of platinum resistance in ovarian cancer. Oncotarget 2017;8:11917–36.PubMedWeb of ScienceGoogle Scholar

  • 20.

    Nakamura K, Sawada K, Yoshimura A, Kinose Y, Nakatsuka E, Kimura T. Clinical relevance of circulating cell-free microRNAs in ovarian cancer. Mol Cancer 2016;15:48.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 21.

    Momen-Heravi F. Isolation of extracellular vesicles by ultracentrifugation. Methods Mol Biol 2017;1660:25–32.Web of SciencePubMedGoogle Scholar

  • 22.

    Silverberg SG. Histopathologic grading of ovarian carcinoma: a review and proposal. Int J Gynecol Pathol 2000;19:7–15.CrossrefPubMedGoogle Scholar

  • 23.

    FIGO Committee on Gynecologic Oncology. Current FIGO staging for cancer of the vagina, fallopian tube, ovary, and gestational trophoblastic neoplasia. Int J Gynaecol Obstet 2009;105:3–4.PubMedGoogle Scholar

  • 24.

    Huang X, Yuan T, Tschannen M, Sun Z, Jacob H, Du M, et al. Characterization of human plasma-derived exosomal RNAs by deep sequencing. BMC Genomics 2013;14:319.PubMedWeb of ScienceGoogle Scholar

  • 25.

    Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2. Nat Methods 2012;9:357–9.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 26.

    Shi J, Dong M, Li L, Liu L, Luz-Madrigal A, Tsonis PA, et al. mirPRo-a novel standalone program for differential expression and variation analysis of miRNAs. Sci Rep 2015;5:14617.Web of SciencePubMedGoogle Scholar

  • 27.

    Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 2014;15:550.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 28.

    Yuan T, Huang X, Woodcock M, Du M, Dittmar R, Wang Y, et al. Plasma extracellular RNA profiles in healthy and cancer patients. Sci Rep 2016;6:19413.CrossrefWeb of SciencePubMedGoogle Scholar

  • 29.

    Thijssen MA, Swinkels DW, Ruers TJ, de Kok JB. Difference between free circulating plasma and serum DNA in patients with colorectal liver metastases. Anticancer Res 2002;22: 421–5.PubMedGoogle Scholar

  • 30.

    Jung M, Klotzek S, Lewandowski M, Fleischhacker M, Jung K. Changes in concentration of DNA in serum and plasma during storage of blood samples. Clin Chem 2003;49:1028–9.PubMedCrossrefGoogle Scholar

  • 31.

    Wang K, Yuan Y, Cho JH, McClarty S, Baxter D, Galas DJ. Comparing the microRNA spectrum between serum and plasma. PLoS One 2012;7:e41561.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 32.

    Shivapurkar N, Vietsch EE, Carney E, Isaacs C, Wellstein A. Circulating microRNAs in patients with hormone receptor-positive, metastatic breast cancer treated with dovitinib. Clin Transl Med 2017;6:37.CrossrefWeb of SciencePubMedGoogle Scholar

  • 33.

    Meng X, Muller V, Milde-Langosch K, Trillsch F, Pantel K, Schwarzenbach H. Diagnostic and prognostic relevance of circulating exosomal miR-373, miR-200a, miR-200b and miR-200c in patients with epithelial ovarian cancer. Oncotarget 2016;7:16923–35.PubMedWeb of ScienceGoogle Scholar

  • 34.

    Melo SA, Luecke LB, Kahlert C, Fernandez AF, Gammon ST, Kaye J, et al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer. Nature 2015;523:177–82.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 35.

    Uratani R, Toiyama Y, Kitajima T, Kawamura M, Hiro J, Kobayashi M, et al. Diagnostic potential of cell-free and exosomal microRNAs in the identification of patients with high-risk colorectal adenomas. PLoS One 2016;11:e0160722.Web of SciencePubMedCrossrefGoogle Scholar

  • 36.

    Schwarzenbach H. Clinical relevance of circulating, cell-free and exosomal microRNAs in plasma and serum of breast cancer patients. Oncol Res Treat 2017;40:423–9.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 37.

    Sodar BW, Kittel A, Paloczi K, Vukman KV, Osteikoetxea X, Szabo-Taylor K, et al. Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection. Sci Rep 2016;6:24316.CrossrefWeb of ScienceGoogle Scholar

  • 38.

    Vickers KC, Palmisano BT, Shoucri BM, Shamburek RD, Remaley AT. MicroRNAs are transported in plasma and delivered to recipient cells by high-density lipoproteins. Nat Cell Biol 2011;13:423–33.Web of SciencePubMedCrossrefGoogle Scholar

  • 39.

    Li L, Xu QH, Dong YH, Li GX, Yang L, Wang LW, et al. MiR-181a upregulation is associated with epithelial-to-mesenchymal transition (EMT) and multidrug resistance (MDR) of ovarian cancer cells. Eur Rev Med Pharmacol Sci 2016;20:2004–10.PubMedGoogle Scholar

  • 40.

    Teng C, Zheng H. Low expression of microRNA-1908 predicts a poor prognosis for patients with ovarian cancer. Oncol Lett 2017;14:4277–81.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 41.

    Ma H, Tian T, Liang S, Liu X, Shen H, Xia M, et al. Estrogen receptor-mediated miR-486-5p regulation of OLFM4 expression in ovarian cancer. Oncotarget 2016;7:10594–605.PubMedWeb of ScienceGoogle Scholar

  • 42.

    Fang G, Liu J, Wang Q, Huang X, Yang R, Pang Y, et al. MicroRNA-223-3p regulates ovarian cancer cell proliferation and invasion by targeting SOX11 expression. Int J Mol Sci 2017;18.PubMedWeb of ScienceGoogle Scholar

  • 43.

    Pink RC, Samuel P, Massa D, Caley DP, Brooks SA, Carter DR. The passenger strand, miR-21-3p, plays a role in mediating cisplatin resistance in ovarian cancer cells. Gynecol Oncol 2015;137:143–51.Web of ScienceCrossrefGoogle Scholar

About the article

Corresponding author: Dr. Jan Dominik Kuhlmann, Department of Gynecology and Obstetrics, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Fetscherstr. 74, 01307 Dresden, Germany, Phone: +49-351-458-2434, Fax: +49-351-458-5844


Received: 2018-09-24

Accepted: 2018-10-09

Published Online: 2018-11-13

Published in Print: 2019-06-26


Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: This study was funded by “VFK Krebsforschung gGmbH”.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 57, Issue 7, Pages 1053–1062, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2018-1048.

Export Citation

©2018 Walter de Gruyter GmbH, Berlin/Boston.Get Permission

Supplementary Article Materials

Comments (0)

Please log in or register to comment.
Log in