Visualization of stem cell activity in pancreatic cancer expansion by direct lineage tracing with live imaging

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease. Although rigorous efforts identified the presence of 'cancer stem cells (CSCs)' in PDAC and molecular markers for them, stem cell dynamics in vivo have not been clearly demonstrated. Here we focused on Doublecortin-like kinase 1 (Dclk1), known as a CSC marker of PDAC. Using genetic lineage tracing with a dual-recombinase system and live imaging, we showed that Dclk1+ tumor cells continuously provided progeny cells within pancreatic intraepithelial neoplasia, primary and metastatic PDAC and PDAC-derived spheroids in vivo and in vitro. Furthermore, genes associated with CSC and epithelial mesenchymal transition were enriched in mouse Dclk1+ and human DCLK1-high PDAC cells. Thus, we provided direct functional evidence for the stem cell activity of Dclk1+ cells in vivo, revealing the essential roles of Dclk1+ cells in expansion of pancreatic neoplasia in all progressive stages.

Data availability

Microarray data have been deposited in GEO under accession codes GSE139167.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Takahisa Maruno

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7060-4104
  2. Akihisa Fukuda

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    For correspondence
    fukuda26@kuhp.kyoto-u.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1940-596X
  3. Norihiro Goto

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  4. Motoyuki Tsuda

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  5. Kozo Ikuta

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  6. Yukiko Hiramatsu

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  7. Satoshi Ogawa

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  8. Yuki Nakanishi

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  9. Yuichi Yamaga

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  10. Takuto Yoshioka

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  11. Kyoichi Takaori

    Hepatobiliary-Pancreatic Surgery and Transplantation, Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  12. Shinji Uemoto

    Hepatobiliary-Pancreatic Surgery and Transplantation, Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  13. Dieter Saur

    Internal Medicine II, Klinikum rechts der Isar Technische Universität München, München, Germany
    Competing interests
    The authors declare that no competing interests exist.
  14. Tsutomu Chiba

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  15. Hiroshi Seno

    Gastroenterology and Hepatology, Kyoto University Graduate School of Medicine, Kyoto, Japan
    For correspondence
    seno@kuhp.kyoto-u.ac.jp
    Competing interests
    The authors declare that no competing interests exist.

Funding

Grants-in-Aid KAKENHI (26293173)

  • Hiroshi Seno

Naito Foundation (N/A)

  • Hiroshi Seno

Princess Takamatsu Cancer Research Fund (13-24514)

  • Tsutomu Chiba

Princess Takamatsu Cancer Research Fund (17-24924)

  • Hiroshi Seno

Takeda Science Foundation (201749741)

  • Hiroshi Seno

Uehara Memorial Foundation (201720143)

  • Hiroshi Seno

Mochida Foundation (201356)

  • Tsutomu Chiba

Mochida Foundation (2017bvAg)

  • Hiroshi Seno

Mitsubishi Foudation (281119)

  • Hiroshi Seno

Mitsubishi Foudation (201910037)

  • Hiroshi Seno

European Research Council (648521)

  • Dieter Saur

Grants-in-Aid KAKENHI (15H06334)

  • Takahisa Maruno

Deutsche Forschungsgemeinschaft (1374/4-2)

  • Dieter Saur

Grants-in-Aid KAKENHI (16K09394)

  • Akihisa Fukuda

Grants-in-Aid KAKENHI (16K15427)

  • Hiroshi Seno

Grants-in-Aid KAKENHI (17H04157)

  • Hiroshi Seno

Grants-in-Aid KAKENHI (19H03639)

  • Akihisa Fukuda

apan Agency for Medical Research and Development (19cm0106142h0002)

  • Hiroshi Seno

apan Agency for Medical Research and Development (19cm6010022h0002)

  • Akihisa Fukuda

Kobayashi Foundation for Cancer Research (N/A)

  • Hiroshi Seno

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Kay F Macleod, University of Chicago, United States

Ethics

Animal experimentation: All animal experiments were approved by the animal research committee of the Kyoto University and performed in accordance with Japanese government regulations. All surgery was performed under Isoflurane anesthesia, and every effort was made to minimize suffering.

Human subjects: Surgically resected specimens of pancreatic cancer tissues were obtained from patients who had been admitted to Kyoto University Hospital. Written informed consent was obtained from all patients and study protocol (#G1200-1) was approved by Ethics Committee of Kyoto University Hospital.

Version history

  1. Received: January 13, 2020
  2. Accepted: November 24, 2020
  3. Accepted Manuscript published: January 4, 2021 (version 1)
  4. Version of Record published: January 11, 2021 (version 2)

Copyright

© 2021, Maruno et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Takahisa Maruno
  2. Akihisa Fukuda
  3. Norihiro Goto
  4. Motoyuki Tsuda
  5. Kozo Ikuta
  6. Yukiko Hiramatsu
  7. Satoshi Ogawa
  8. Yuki Nakanishi
  9. Yuichi Yamaga
  10. Takuto Yoshioka
  11. Kyoichi Takaori
  12. Shinji Uemoto
  13. Dieter Saur
  14. Tsutomu Chiba
  15. Hiroshi Seno
(2021)
Visualization of stem cell activity in pancreatic cancer expansion by direct lineage tracing with live imaging
eLife 10:e55117.
https://doi.org/10.7554/eLife.55117

Share this article

https://doi.org/10.7554/eLife.55117

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