Desialylation of Sonic-Hedgehog by Neu2 Inhibits Its Association with Patched1 Reducing Stemness-Like Properties in Pancreatic Cancer Sphere-forming Cells

Cancer stem cells (CSCs) are crucial regulators of tumor recurrence/progression. The maintenance of CSCs is dependent on aberrant activation of various pathways, including Hedgehog. Prevalent sialylations contribute to aggressiveness in CSCs. Here, we have addressed the role of sialylation in regulating stemness-like properties of pancreatic cancer sphere-forming cells (PCS) through modulation of the Hedgehog (Hh) pathway. The status of CD133/CD44/surface-sialylation was checked by flow cytometry and effects of Neu2 overexpression in PCS were compared using qPCR, immunoblotting, co-immunoprecipitation and also by colony-formation assays. The work was also validated in a xenograft model after Neu2 overexpression. Neu2 and Shh status in patient tissues were examined by immunohistochemistry. PCS showed higher Hh-pathway activity and sialylation with reduced cytosolic-sialidase (Neu2). Neu2 overexpression caused desialylation of Shh, thereby reducing Shh-Patched1 binding thus causing decreased Hh-pathway activity with lower expression of Snail/Slug/CyclinD1 leading to reduction of stemness-like properties. Neu2-overexpression also induced apoptosis in PCS. Additionally, Neu2-overexpressed PCS demonstrated lower mTORC2 formation and inhibitory-phosphorylation of Gsk3β, reflecting a close relationship with reduced Hh pathway. Moreover, both Neu2 and Rictor (a major component of mTORC2) co-transfection reduced stem cell markers and Hh-pathway activity in PCS. Neu2-overexpressed tumors showed reduction in tumor mass with downregulation of stem cell markers/Shh/mTOR and upregulation of Bax/Caspase8/Caspase3. Thus, we established that reduced sialylation by Neu2 overexpression leads to decreased stemness-like properties by desialylation of Shh, which impaired its association with Patched1 thereby inhibiting the Hh pathway. All these may be responsible for enhanced apoptosis in Neu2-overexpressed PCS.

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Cyclopamine reverts acquired chemoresistance and down-regulates cancer stem cell markers in pancreatic cancer cell lines

Swiss Medical Weekly ◽

10.4414/smw.2011.13208 ◽

2011 ◽

Cited By ~ 2

Author(s):

J Yao ◽

Y An ◽

JS Wie ◽

ZL Ji ◽

ZP Lu ◽

...

Keyword(s):

Pancreatic Cancer ◽

Stem Cell ◽

Cancer Stem Cell ◽

Cell Lines ◽

Cancer Cell ◽

Pancreatic Cancer Cell ◽

Cancer Cell Lines ◽

Stem Cell Markers ◽

Cell Markers ◽

Cancer Stem Cell Markers

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Discovery and 3D imaging of a novel ΔNp63-expressing basal cell type in human pancreatic ducts with implications in disease

10.1101/2020.08.20.259317 ◽

2020 ◽

Author(s):

Sandrina Martens ◽

Mathias Van Bulck ◽

Katarina Coolens ◽

Hediel Madhloum ◽

Farzad Esni ◽

...

Keyword(s):

Pancreatic Cancer ◽

Stem Cell ◽

Chronic Pancreatitis ◽

Basal Cell ◽

3D Imaging ◽

Stem Cell Markers ◽

List Type ◽

Human Pancreas ◽

Basal Cells ◽

Cell Markers

SUMMARYObjectiveAn aggressive basal-like molecular subtype of pancreatic ductal adenocarcinoma (PDAC) exists, driven by ΔNp63. In other epithelia, ΔNp63+ basal cells have stem cell capacity and can be at the origin of tumors. In the pancreas, basal cells have not been identified.DesignWe assessed basal cell markers in human and mouse pancreas, chronic pancreatitis and PDAC, and developed a 3D imaging protocol (FLIP-IT) to study sizeable samples at single cell resolution. We generated organoid cultures of ducts from Sox9-eGFP reporter mice.ResultsIn normal human pancreas, rare ΔNp63+ cells exist in ducts that expand in chronic pancreatitis. ΔNp63+ cells express KRT19 and canonical basal markers (KRT5, KRT14 and S100A2) but lack markers of duct cells such as CA19.9 and SOX9. In addition, ΔNp63+ cells pertain to a niche of cells expressing gastrointestinal stem cell markers. 3D views of the ductal tree in formalin fixed paraffin embedded samples show that basal cells are localized on the basal membrane of medium to large ducts and expand as multilayer dome-like structures in chronic pancreatitis. In mice, ΔNp63 expression is induced when culturing organoids from Sox9-low ductal cells but could not be found in normal pancreas nor in models of pancreatitis or pancreatic cancer.ConclusionWe discovered a novel ductal cell population in normal human pancreas similar to basal cells in other tissues. Using FLIP-IT, we provide unprecedented 3D visualization of these cells in archival clinical specimens. ΔNp63+ cells may play an important role in pancreatic tissue regeneration and cancer.SUMMARY BOXWhat is already known about this subject?ΔNp63 has a central role in determining the basal-like subtype of pancreatic ductal adenocarcinoma (PDAC).Different to other tissues with basal cancers, the normal pancreas reportedly does not contain (ΔNp63-expressing) basal cells.Current protocols face severe limitations for marker-based identification and 3D imaging of individual (rare) cells in archival pancreatic samples.What are the new findings?We report a rare and atypical pancreatic duct cell that expresses ΔNp63, other basal cell markers and g.i. stem cell markers.The number of these basal cells increases in diseases such as chronic pancreatitis and pancreatic cancer.We provide an easy to implement protocol for 3D clearing and high-resolution imaging of sizeable samples of (fresh or FFPE) human pancreas or an entire mouse pancreas.Except after culturing medium to large ducts as organoids, we fail to detect basal cells in mouse experimental pancreatic models.How might it impact on clinical practice in the foreseeable future?Extrapolating from knowledge in other organs, basal cells in the pancreas may have a stem cell/progenitor role, including in diseases such as (basal) pancreatic cancer.Use of the 3D imaging protocol in archival clinical specimens will allow unprecedented insights in pancreatic histopathology.For above mentioned diseases, we caution for findings in experimental mouse models that may not (fully) recapitulate the etiopathogenesis.

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Do pancreatic cancer (PDA) stem cell markers predict biologic behavior?

Journal of Clinical Oncology ◽

10.1200/jco.2016.34.15_suppl.4112 ◽

2016 ◽

Vol 34 (15_suppl) ◽

pp. 4112-4112

Author(s):

Ibrahim Halil Sahin ◽

Mesruh Turkekul ◽

Marinela Capanu ◽

Gokce Askan ◽

Breanna McMahon ◽

...

Keyword(s):

Pancreatic Cancer ◽

Stem Cell ◽

Stem Cell Markers ◽

Cell Markers

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Role of cancer stem cell markers ALDH1, BCL11B, BMI-1, and CD44 in the prognosis of advanced HNSCC

10.1055/s-0040-1710997 ◽

2020 ◽

Author(s):

K Sharaf ◽

J Kitz ◽

M Canis ◽

M Jakob

Keyword(s):

Stem Cell ◽

Cancer Stem Cell ◽

Stem Cell Markers ◽

Cell Markers ◽

Cancer Stem Cell Markers

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Hepatoblastoma Survival and the Prognostic Role of Cancer Stem Cell Markers

International Journal of Cancer Therapy and Oncology ◽

10.14319/ijcto.0201.1 ◽

2014 ◽

Vol 2 (1) ◽

Author(s):

Mohamed Fawzy ◽

Abeer Bahnassy ◽

Mohamed El-Wakil ◽

Ahmed Abdel-Sayed

Keyword(s):

Stem Cell ◽

Cancer Stem Cell ◽

Stem Cell Markers ◽

Prognostic Role ◽

Cell Markers ◽

Cancer Stem Cell Markers

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Mouse models of pancreatic cancer induced by chronic pancreatitis and smoking.

Journal of Clinical Oncology ◽

10.1200/jco.2014.32.3_suppl.229 ◽

2014 ◽

Vol 32 (3_suppl) ◽

pp. 229-229

Author(s):

M. Edderkaoui ◽

Paul J. Grippo ◽

Y. Ouhaddi ◽

H. Benhaddou ◽

S. Xu ◽

...

Keyword(s):

Pancreatic Cancer ◽

Stem Cell ◽

Chronic Pancreatitis ◽

Transgenic Mice ◽

Mouse Models ◽

Tobacco Smoke ◽

Stellate Cell ◽

Stem Cell Markers ◽

Lesion Formation ◽

Cell Markers

229 Background: Chronic pancreatitis (CP) and smoking are two major risk factors for pancreatic cancer (PaCa). Here we developed mouse models of PaCa by exposing mice to CP and/or smoking. Methods: Wild type (WT) and Pdx1-Cre;LSL-Kras transgenic mice were subjected to 7-hourly cerulein injections (50µg/kg) twice a week for 3 weeks to induce CP. Another group of mice was exposed to tobacco smoke (80mg/m3) in chambers for 5days/week during 6 weeks. A third group was exposed to both treatments. Mice were then sacrificed and pancreatic tissue analyzed. Pancreatic lesions, proliferation, fibrosis, inflammation, EMT and stem cell markers were analyzed by immunohistochemistry and Western. Results: CP significantly increased the number and stage of pancreatic neoplastic (PanIN) lesions with fibrosis, stellate cell activation and inflammation in transgenic mice. These changes were significantly less in WT mice. EMT markers (decrease in E-Cadherin and increase in vimentin protein levels) were stimulated by CP in both WT and transgenic mice. Similarly, smoking stimulated PanIN lesion formation, fibrosis and inflammation in transgenic mice, but to a lesser extent than CP model. Expression of EMT and stem cell markers was greater in the smoking model compared to CP. Combination of CP and smoking induced greater stimulation of PanIN lesion formation, fibrosis and inflammation compared to either treatment alone in transgenic mice. Conclusions: The results indicate that both CP and tobacco smoke promote PaCa progression in transgenic mice. The combination of CP and smoking has greater effects than either treatment alone. EMT and stem cell markers developed in WT as well as in transgenic mice with smoking suggesting that smoking has effects independent of Kras on cell transformation.

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