Skip to main content

Advertisement

SpringerLink
Log in

Pancreatic Juice Exosomal MicroRNAs as Biomarkers for Detection of Pancreatic Ductal Adenocarcinoma

  • Pancreatic Tumors
  • Published:
Annals of Surgical Oncology Aims and scope Submit manuscript

Abstract

Background

Pancreatic ductal adenocarcinoma (PDAC) is a lethal neoplasm because of difficulties in early detection. Several studies have recently suggested that exosomes may have potential as novel biomarkers. This study aimed to isolate exosomes from pancreatic juice and to investigate whether exosomal microRNAs (ex-miRs) could be used as biomarkers for PDAC.

Methods

Pancreatic juice was collected from patients with PDAC and chronic pancreatitis (CP) by endoscopic retrograde pancreatography. Exosomes were extracted by ultracentrifugation. The presence of exosomes was confirmed by electron microscopy and Western blotting using anti-CD63, -CD81, and -TSG101 antibodies. Relative levels of ex-miR-21 and ex-miR-155 were quantified and compared between PDAC and CP patients.

Results

A total of 35 pancreatic juice samples (27 PDAC and 8 CP) were collected. Relative levels of both ex-miR-21 and ex-miR-155 were significantly higher in PDAC patients compared with CP patients (p < 0.001 and p = 0.008, respectively). By contrast, no significant difference was apparent in relative levels of miR-21 and miR-155 in whole pancreatic juice from PDAC patients compared with CP patients (p = 0.08 and p = 0.61, respectively). Ex-miR-21 and ex-miR-155 levels discriminated PDAC patients from CP patients with area under the curve values of 0.90 and 0.89, respectively. The accuracies of ex-miR-21 levels, ex-miR-155 levels, and pancreatic juice cytology were 83%, 89%, and 74%, respectively. When combining the results of ex-miR profiling with pancreatic juice cytology, the accuracy was improved to 91%.

Conclusions

We successfully extracted exosomes from pancreatic juice. Ex-miRs, including ex-miR-21 and ex-miR-155, in pancreatic juice may be developed as biomarkers for PDAC.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

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

    Article  PubMed  Google Scholar 

  2. Ahmed K, Sussman JJ, Wang J, et al. A case of EUS-guided FNA-related pancreatic cancer metastasis to the stomach. Gastrointest Endosc 2011;74:231–3.

    Article  PubMed  Google Scholar 

  3. Chong A, Venugopal K, Segarajasingam D, et al. Tumor seeding after EUS-guided FNA of pancreatic tail neoplasia. Gastrointest Endosc 2011;74:933–5.

    Article  PubMed  Google Scholar 

  4. Kanno A, Masamune A, Hanada K, et al. Multicenter study of early pancreatic cancer in Japan. Pancreatology 2018;18:61–67.

    Article  PubMed  Google Scholar 

  5. Kimura H, Ohtsuka T, Matsunaga T, et al. Predictors and diagnostic strategies for early-stage pancreatic ductal adenocarcinoma: a retrospective study. Pancreas 2015;44:1148–54.

    Article  CAS  PubMed  Google Scholar 

  6. Ohtsuka T, Tamura K, Ideno N, et al. Role of ERCP in the era of EUS-FNA for preoperative cytological confirmation of resectable pancreatic ductal adenocarcinoma. Surg Today 2014;44:1887–92.

    Article  PubMed  Google Scholar 

  7. Bartel DP. MicroRNAs: target recognition and regulatory functions. Cell 2009;136:215–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Chen X, Ba Y, Ma L, et al. Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res 2008;18:997–1006.

    Article  CAS  PubMed  Google Scholar 

  9. Xu J, Cao Z, Liu W, et al. Plasma miRNAs Effectively distinguish patients with pancreatic cancer from controls: a multicenter study. Ann Surg 2016;263:1173-9.

    Article  PubMed  Google Scholar 

  10. Li A, Yu J, Kim H, et al. MicroRNA array analysis finds elevated serum miR-1290 accurately distinguishes patients with low-stage pancreatic cancer from healthy and disease controls. Clin Cancer Res 2013;19:3600–10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Abue M, Yokoyama M, Shibuya R, et al. Circulating miR-483-3p and miR-21 is highly expressed in plasma of pancreatic cancer. Int J Oncol 2015;46:539–47.

    Article  CAS  PubMed  Google Scholar 

  12. Schultz NA, Dehlendorff C, Jensen BV, et al. MicroRNA biomarkers in whole blood for detection of pancreatic cancer. JAMA 2014;311:392–404.

    Article  CAS  PubMed  Google Scholar 

  13. Silverman JM, Reiner NE. Exosomes and other microvesicles in infection biology: organelles with unanticipated phenotypes. Cell Microbiol 2011;13:1–9.

    Article  CAS  PubMed  Google Scholar 

  14. Que R, Ding G, Chen J, et al. Analysis of serum exosomal microRNAs and clinicopathologic features of patients with pancreatic adenocarcinoma. World J Surg Oncol 2013;11:219.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Madhavan B, Yue S, Galli U, et al. Combined evaluation of a panel of protein and miRNA serum-exosome biomarkers for pancreatic cancer diagnosis increases sensitivity and specificity. Int J Cancer 2015;136:2616–27.

    Article  CAS  PubMed  Google Scholar 

  16. Xu YF, Hannafon BN, Zhao YD, et al. Plasma exosome miR-196a and miR-1246 are potential indicators of localized pancreatic cancer. Oncotarget 2017;8:77028–77040.

    PubMed  PubMed Central  Google Scholar 

  17. Mikamori M, Yamada D, Eguchi H, et al. MicroRNA-155 controls exosome synthesis and promotes gemcitabine resistance in pancreatic ductal adenocarcinoma. Sci Rep 2017;7:42339.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Lai X, Wang M, McElyea SD, et al. A microRNA signature in circulating exosomes is superior to exosomal glypican-1 levels for diagnosing pancreatic cancer. Cancer Lett 2017;393:86–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Takahasi K, Iinuma H, Wada K, et al. Usefulness of exosome-encapsulated microRNA-451a as a minimally invasive biomarker for prediction of recurrence and prognosis in pancreatic ductal adenocarcinoma. J Hepatobiliary Pancreat Sci 2018;25:155–161.

    Article  PubMed  Google Scholar 

  20. Goto T, Fujiya M, Konishi H, et al. An elevated expression of serum exosomal microRNA-191, -21, -451a of pancreatic neoplasm is considered to be efficient diagnostic marker. BMC Cancer 2018;18:116.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Sadakari Y, Ohtsuka T, Ohuchida K, et al. MicroRNA expression analyses in preoperative pancreatic juice samples of pancreatic ductal adenocarcinoma. JOP 2010;11:587–92.

    PubMed  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Li L, Masica D, Ishida M, et al. Human bile contains microRNA-laden extracellular vesicles that can be used for cholangiocarcinoma diagnosis. Hepatology 2014;60:896–907.

    Article  CAS  PubMed  Google Scholar 

  24. Lotvall J, Hill AF, Hochberg F, et al. Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. J Extracell Vesicles 2014;3:26913.

    Article  PubMed  Google Scholar 

  25. Wang J, Chen J, Chang P, 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.

    Article  CAS  Google Scholar 

  26. Zheng J, Hernandez JM, Doussot A, et al. Extracellular matrix proteins and carcinoembryonic antigen-related cell adhesion molecules characterize pancreatic duct fluid exosomes in patients with pancreatic cancer. HPB (Oxford) 2018;20(7):597–604.

    Article  Google Scholar 

  27. Severino V, Dumonceau JM, Delhaye M, et al. Extracellular vesicles in bile as markers of malignant biliary stenoses. Gastroenterology 2017;153:495–504.e8.

    Google Scholar 

  28. Pan X, Wang ZX, Wang R. MicroRNA-21: a novel therapeutic target in human cancer. Cancer Biol Ther 2010;10:1224–32.

    Article  CAS  PubMed  Google Scholar 

  29. Gironella M, Seux M, Xie MJ, et al. Tumor protein 53-induced nuclear protein 1 expression is repressed by miR-155, and its restoration inhibits pancreatic tumor development. Proc Natl Acad Sci USA 2007;104:16170–5.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Suenaga M, Sadakari Y, Almario JA, et al. Using an endoscopic distal cap to collect pancreatic fluid from the ampulla (with video). Gastrointest Endosc 2017;86:1152–1156.e2.

    Google Scholar 

  31. Yu J, Sadakari Y, Shindo K, et al. Digital next-generation sequencing identifies low-abundance mutations in pancreatic juice samples collected from the duodenum of patients with pancreatic cancer and intraductal papillary mucinous neoplasms. Gut 2017;66:1677–1687.

    Article  CAS  PubMed  Google Scholar 

  32. Suenaga M, Yu J, Shindo K, et al. Pancreatic juice mutation concentrations can help predict the grade of dysplasia in patients undergoing pancreatic surveillance. Clin Cancer Res 2018;24(12):2963–2974.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This study was supported by a Grant-in-Aid for Scientific Research (Grant numbers 15K10186, 17K10637, 17K10636) and a JFE (The Japanese Foundation for Research and Promotion of Endoscopy) grant.

Author information

Authors and Affiliations

  1. Department of Surgery and Oncology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    So Nakamura MD, Yoshihiko Sadakari MD, PhD, Takao Ohtsuka MD, PhD, Takafumi Okayama MD, Yohei Nakashima MD, Yoshitaka Gotoh MD, Yasuhisa Mori MD, PhD, Kohei Nakata MD, PhD, Yoshihiro Miyasaka MD, PhD & Masafumi Nakamura MD, PhD, FACS

  2. Department of Anatomic Pathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    Kiyoshi Saeki MD & Yoshinao Oda MD, PhD

  3. Department of Cancer Therapy and Research, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    Hideya Onishi MD, PhD

  4. Departments of Pathology, Medicine, and Oncology, The Sol Goldman Pancreatic Cancer Research Center, The Johns Hopkins Medical Institutions, Baltimore, MD, USA

    Michael Goggins MB, MD

Authors
  1. So Nakamura MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yoshihiko Sadakari MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takao Ohtsuka MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takafumi Okayama MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yohei Nakashima MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yoshitaka Gotoh MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kiyoshi Saeki MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yasuhisa Mori MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Kohei Nakata MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yoshihiro Miyasaka MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Hideya Onishi MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Yoshinao Oda MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Michael Goggins MB, MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Masafumi Nakamura MD, PhD, FACS
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

SN, TO, YN, and YG contributed to establishing the study population database and performed data extraction and analysis. YS, TO, YM, KN, YM, HO, MG, and MN contributed to study conception and design and performed the pancreatectomy and follow-up of the study population. KS and YO made the pathological diagnoses. SN wrote the manuscript. YS and TO contributed to the interpretation of results and manuscript revision. All authors discussed the results and commented on the manuscript. YS, TO, YM, KN, YM, HO, MG, and MN provided the final approval for this article.

Corresponding author

Correspondence to Masafumi Nakamura MD, PhD, FACS.

Ethics declarations

Disclosure

So Nakamura, Yoshihiko Sadakari, Takao Ohtsuka, Takafumi Okayama, Yohei Nakashima, Yoshitaka Gotoh, Kiyoshi Saeki, Yasuhisa Mori, Kohei Nakata, Yoshihiro Miyasaka, Hideya Onishi, Yoshinao Oda, Michael Goggins, and Masafumi Nakamura disclosed no financial relationships relevant to this publication.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic Supplementary Material

Below is the link to the electronic supplementary material.

Fig. S1

The representative result of Bioanalyzer 2100. Total RNA extracted from the exosomes isolated from pancreatic juice was analyzed using Bioanalyzer 2100. The data showed the enrichment of small RNAs in exosome fraction. (TIF 207 kb)

Fig. S2

Stability of miRNA expression in exosomes. (a) Levels of miR-21 were stable in both exosomes (ex-miR-21) and whole pancreatic juice (free-miR-21) stored at room temperature. (b) Levels of ex-miR-21 were stable at 37 °C, while levels of free-miR-21 were degraded over time. (TIF 133 kb)

Fig. S3

Relative expression levels of (a) miR-21 and (b) miR-155 in formalin-fixed paraffin embedded (FFPE) tissue samples, whole pancreatic juice, and exosomes obtained randomly from five PDAC patients. The relative expression levels of exosomal miRs were equivalent or higher than those of tissue samples, while those of miRs in whole pancreatic juice tended to be lower than tissue samples. (TIF 147 kb)

Supplementary material 4 (docx 24 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nakamura, S., Sadakari, Y., Ohtsuka, T. et al. Pancreatic Juice Exosomal MicroRNAs as Biomarkers for Detection of Pancreatic Ductal Adenocarcinoma. Ann Surg Oncol 26, 2104–2111 (2019). https://doi.org/10.1245/s10434-019-07269-z

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1245/s10434-019-07269-z

Search

Navigation