Isolation and in Vitro Culture of Rare Cancer Stem Cells from Patient-Derived Xenografts of Pancreatic Ductal Adenocarcinoma

Abstract Pancreatic ductal adenocarcinoma (PDAC), the fourth leading cause of cancer death, has a 5-year survival rate of approximately 7–9%. The ineffectiveness of anti-PDAC therapies is believed to be due to the existence of a subpopulation of tumor cells known as cancer stem cells (CSCs), which are functionally plastic, and have exclusive tumorigenic, chemoresistant and metastatic capacities. Herein, we describe a 2D in vitro system for long-term enrichment of pancreatic CSCs that is amenable to biological and CSC-specific studies. By changing the carbon source from glucose to galactose in vitro, we force PDAC cells to utilize OXPHOS, resulting in enrichment of CSCs defined by increased CSC biomarker and pluripotency gene expression, greater tumorigenic potential, induced but reversible quiescence, increased OXPHOS activity, enhanced invasiveness, and upregulated immune evasion properties. This CSC enrichment method can facilitate the discovery of new CSC-specific hallmarks for future development into targets for PDAC-based therapies.

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Induction of Lysosome Membrane Permeabilization as a Therapeutic Strategy to Target Pancreatic Cancer Stem Cells

Cancers ◽

10.3390/cancers12071790 ◽

2020 ◽

Vol 12 (7) ◽

pp. 1790 ◽

Cited By ~ 3

Author(s):

Timothy P. Cash ◽

Sonia Alcalá ◽

María del Rosario Rico-Ferreira ◽

Elena Hernández-Encinas ◽

Jennifer García ◽

...

Keyword(s):

Pancreatic Cancer ◽

Stem Cells ◽

Cancer Stem Cells ◽

Therapeutic Strategy ◽

Membrane Permeabilization ◽

Ductal Adenocarcinoma ◽

Chemical Library ◽

Specific Chemical ◽

Pancreatic Cancer Stem Cells

Despite significant efforts to improve pancreatic ductal adenocarcinoma (PDAC) clinical outcomes, overall survival remains dismal. The poor response to current therapies is partly due to the existence of pancreatic cancer stem cells (PaCSCs), which are efficient drivers of PDAC tumorigenesis, metastasis and relapse. To find new therapeutic agents that could efficiently kill PaCSCs, we screened a chemical library of 680 compounds for candidate small molecules with anti-CSC activity, and identified two compounds of a specific chemical series with potent activity in vitro and in vivo against patient-derived xenograft (PDX) cultures. The anti-CSC mechanism of action of this specific chemical series was found to rely on induction of lysosomal membrane permeabilization (LMP), which is likely associated with the increased lysosomal mass observed in PaCSCs. Using the well characterized LMP-inducer siramesine as a tool molecule, we show elimination of the PaCSC population in mice implanted with tumors from two PDX models. Collectively, our approach identified lysosomal disruption as a promising anti-CSC therapeutic strategy for PDAC.

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Inhibition Of The Notch Signaling Pathway Targets Cancer Stem Cells In Pancreatic Ductal Adenocarcinoma

Journal of Surgical Research ◽

10.1016/j.jss.2010.11.354 ◽

2011 ◽

Vol 165 (2) ◽

pp. 303 ◽

Cited By ~ 1

Author(s):

E.N. Proctor ◽

J. Wu ◽

M.J. Hynes ◽

J. Dosch ◽

D.M. Simeone

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

Notch Signaling ◽

Pancreatic Ductal Adenocarcinoma ◽

Signaling Pathway ◽

Notch Signaling Pathway ◽

Ductal Adenocarcinoma

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Cancer stem cells and mesenchymal stem cells in pancreatic ductal adenocarcinoma

Pathologia ◽

10.14739/2310-1237.2019.1.166476 ◽

2019 ◽

Vol 0 (1) ◽

Author(s):

V. O. Tumanskyi ◽

I. S. Kovalenko

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cancer Stem Cells ◽

Pancreatic Ductal Adenocarcinoma ◽

Ductal Adenocarcinoma

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Moonlighting Role of PEDF in Breast Cancer

10.21203/rs.3.rs-93992/v1 ◽

2020 ◽

Author(s):

Carmen Gil-Gas ◽

Marta Sánchez-Díez ◽

Paloma Honrubia-Gómez ◽

Jose Luis Sánchez-Sánchez ◽

Carmen Belen Alvarez-Simón ◽

...

Keyword(s):

Breast Cancer ◽

Stem Cells ◽

Stem Cell ◽

Cancer Stem Cells ◽

Cancer Patient ◽

In Vitro Culture ◽

Screening Tests ◽

Stem Cell Markers ◽

Self Renewal

Abstract Background: Breast cancer is the leading cause of death among females in developed countries. Although the implementation of screening tests and the development of new therapies has increased the probability of remission, relapse rates still remain high. Numerous studies have indicated the connection between cancer initiating cells and slow cellular cycle cells, identified by their capacity to retain long labelling (LT+). Methods: We have designed a transgenic protein consisting in the C-terminal part of this protein, which acts by blocking endogenous PEDF in culture cell assays. Present work is based in doses-response in vitro assays as well as flow cytometry analysis of surface markers and cell cycle kinetic study of the tumour initiating cells.Results: In this study we show that this type of cells is present not only in cancer cell lines but also in cancer cells from patients with metastatic and advanced stage tumours. We also present new assays showing how stem cell self-renewal modulating proteins, such as PEDF, can modify the properties, expression of markers, and carcinogenicity of cancer stem cells. This protein has been involved in self-renewal in adult stem cells and has been described as anti-tumoral because of its anti-angiogenic effect. However, we show that PEDF enhances resistance in breast cancer patient cells in vitro culture by favoring a slow cellular cycle population (LT+). The PEDF signalling pathway could be a useful tool for controlling cancer stem cells self-renewal, and therefore control patient relapse. Conclusions: We demonstrate that it is possible to interfere with the self-renewal capacity of cancer stem cells, induce anoikis in vivo, and reduce resistance against Docetaxel treatment in cancer patient cells in vitro culture. We have also demonstrated that this PEDF modified protein produces a significant decrease in cancer stem cell markers. All these properties make this protein a potential application in clinical cancer therapies via co-administration with chemotherapy for relapse cancer treatment.

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Expression of stem cell markers in pancreatic ductal adenocarcinoma and clinical relevance.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.e15058 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. e15058-e15058

Author(s):

Kalliopi Andrikou ◽

Luca Faloppi ◽

Cristian Loretelli ◽

Alessandra Mandolesi ◽

Italo Bearzi ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cancer Stem Cells ◽

Pancreatic Ductal Adenocarcinoma ◽

Univariate Analysis ◽

Human Pancreatic Cancer ◽

Ductal Adenocarcinoma ◽

Small Subset ◽

Stem Cell Markers ◽

Cell Markers

e15058 Background: Emerging evidence has suggested that malignant tumors are heterogeneous and that are composed of a small subset of distinct cancer cells (usually defined by cell surface marker expression) that are responsible for tumor initiation and propagation, termed cancer stem cells. These cancer stem cells (CSCs), or tumor-initiating cells (TICs), exhibit properties of normal stem cells and are associated with resistance to current therapies. It was found that human pancreatic cancer has shown a population of cancer stem cells that have aberrantly activated developmental signaling pathways, are resistant to standard chemotherapy and radiation, and have up-regulated signaling cascades that are integral for tumor metastasis. The aim of our study is to investigate the prognostic implication of cancer stem cell markers in pancreatic ductal adenocarcinoma. Methods: In 43 histological samples of pancreatic ductal adenocarcinoma were performed molecular biology assessment of CD 24, CD44, CD133, CD166, OCT3/4, LGR5. Results: At univariate analysis patients with an overexpression of CD44, CD133, OCT3/4 showed a worse prognosis in terms of overall survival (respectively: p=0.031; p=0.014; p=0.001). At multivariate analysis OCT3/4 resulted to be independent factor influencing outcome (HR=0.23). Patients with overexpression of all factors seem to have a worse OS compared to patients expressing only two, one or none (8.6 vs. 15.6 vs. 18.0 vs. 59.9 mts; p=0.006). Conclusions: Our data suggest that the presence of cancer stem cells could be linked with tumor aggressiveness and patients’ survival. This finding could drive therapeutic decision towards less or more intensive treatment.

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