Visualization of stem cell activity in pancreatic cancer expansion by direct lineage tracing with live imaging
- Kyoto University Graduate School of Medicine, Japan
- Klinikum rechts der Isar Technische Universität München, Germany
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.
-
Gene expression profiles of Dclk1+ and Dclk1- PDAC cellsNCBI Gene Expression Omnibus, GSE139167.
-
Expression data from Mayo Clinic Pancreatic Tumor and Normal samplesNCBI Gene Expression Omnibus, GSE16515.
-
Integrative Survival-Based Molecular Profiling of Human Pancreatic CancerNCBI Gene Expression Omnibus, GSE32676.
Article and author information
Author details
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.
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.
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.
Metrics
-
- 3,582
- views
-
- 507
- downloads
-
- 21
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
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
Further reading
-
- Cancer Biology
Glioblastomas are aggressive brain tumors with dismal prognosis. One of the main bottlenecks for developing more effective therapies for glioblastoma stems from their histologic and molecular heterogeneity, leading to distinct tumor microenvironments and disease phenotypes. Effectively characterizing these features would improve the clinical management of glioblastoma. Glucose flux rates through glycolysis and mitochondrial oxidation have been recently shown to quantitatively depict glioblastoma proliferation in mouse models (GL261 and CT2A tumors) using dynamic glucose-enhanced (DGE) deuterium spectroscopy. However, the spatial features of tumor microenvironment phenotypes remain hitherto unresolved. Here, we develop a DGE Deuterium Metabolic Imaging (DMI) approach for profiling tumor microenvironments through glucose conversion kinetics. Using a multimodal combination of tumor mouse models, novel strategies for spectroscopic imaging and noise attenuation, and histopathological correlations, we show that tumor lactate turnover mirrors phenotype differences between GL261 and CT2A mouse glioblastoma, whereas recycling of the peritumoral glutamate-glutamine pool is a potential marker of invasion capacity in pooled cohorts, linked to secondary brain lesions. These findings were validated by histopathological characterization of each tumor, including cell density and proliferation, peritumoral invasion and distant migration, and immune cell infiltration. Our study bodes well for precision neuro-oncology, highlighting the importance of mapping glucose flux rates to better understand the metabolic heterogeneity of glioblastoma and its links to disease phenotypes.
-
- Cancer Biology
- Medicine
A doctoral-level internship program was developed at the University of North Carolina at Chapel Hill with the intent to create customizable experiential learning opportunities for biomedical trainees to support career exploration, preparation, and transition into their postgraduate professional roles. We report the outcomes of this program over a 5-year period. During that 5-year period, 123 internships took place at over 70 partner sites, representing at least 20 academic, for-profit, and non-profit career paths in the life sciences. A major goal of the program was to enhance trainees’ skill development and expertise in careers of interest. The benefits of the internship program for interns, host/employer, and supervisor/principal investigator were assessed using a mixed-methods approach, including surveys with closed- and open-ended responses as well as focus group interviews. Balancing stakeholder interests is key to creating a sustainable program with widespread support; hence, the level of support from internship hosts and faculty members were the key metrics analyzed throughout. We hypothesized that once a successful internship program was implemented, faculty culture might shift to be more accepting of internships; indeed, the data quantifying faculty attitudes support this. Furthermore, host motivation and performance expectations of interns were compared with results achieved, and this data revealed both expected and surprising benefits to hosts. Data suggests a myriad of benefits for each stakeholder group, and themes are cataloged and discussed. Program outcomes, evaluation data, policies, resources, and best practices developed through the implementation of this program are shared to provide resources that facilitate the creation of similar internship programs at other institutions. Program development was initially spurred by National Institutes of Health pilot funding, thereafter, successfully transitioning from a grant-supported model, to an institutionally supported funding model to achieve long-term programmatic sustainability.
