CE-CAM model for evaluating CD133lo Cancer Stem Cells in Retinoblastoma

Abstract BackgroundCancer Stem Cells (CSCs) reported in various tumors, play a crucial role in tumorigenesis and metastasis. Following the efforts to reduce, replace and refine the use of mammalian models, we aimed to establish a short-term xenograft for Retinoblastoma (Rb) to evaluate the tumorigenic and metastatic potential of CD133lo CSCs in Rb Y79 cells, using the well-established chick embryo (CE) model. MethodsTotal and CD133 sorted Rb Y79 cells, labelled with eGFP/CM-Dil tracking dye, were transplanted onto the chorioallantoic membrane (CAM) of day-7 chick embryos and incubated for 7 days. The tumor formation on CAM and metastasis to the embryos were evaluated by confocal microscopy, in-vivo imaging, and histopathology. ResultsY79 cells formed pink-white raised perivascular nodules on the CAM with CD133lo CSCs exhibiting larger nodules when compared to CD133hi cells and total Y79 (p<0.05). In-vivo imaging revealed that the labeled cells metastasized to the embryos with the fluorescent signals visible in the abdominal area, cephalus and the limbs. Histopathologic studies confirmed the presence of tumor cells on the CAM, organs of embryos transplanted with Y79 cells, more so with CD133lo CSCs. ConclusionsThis study highlights that the CE-CAM is a feasible alternative non-mammalian model for evaluating tumorigenicity and metastatic potential of Rb CSCs. The study also provides preliminary evidence that Rb Y79 CD133lo CSCs show higher propensity to form tumor nodules on the CAM and are more invasive than non CSCs, thus, supporting our earlier evidence that they are endowed with CSC properties.

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Cancer Stem Cells and Macrophages: Implications in Tumor Biology and Therapeutic Strategies

Mediators of Inflammation ◽

10.1155/2016/9012369 ◽

2016 ◽

Vol 2016 ◽

pp. 1-15 ◽

Cited By ~ 43

Author(s):

Bruno Sainz ◽

Emily Carron ◽

Mireia Vallespinós ◽

Heather L. Machado

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

Cross Talk ◽

Tumor Biology ◽

Cell Types ◽

Tumor Formation ◽

Self Renewal ◽

Numerous Cell ◽

Key Drivers

Cancer stem cells (CSCs) are a unique subset of cells within tumors with stemlike properties that have been proposed to be key drivers of tumor initiation and progression. CSCs are functionally defined by their unlimited self-renewal capacity and their ability to initiate tumor formationin vivo. Like normal stem cells, CSCs exist in a cellular niche comprised of numerous cell types including tumor-associated macrophages (TAMs) which provides a unique microenvironment to protect and promote CSC functions. TAMs provide pivotal signals to promote CSC survival, self-renewal, maintenance, and migratory ability, and in turn, CSCs deliver tumor-promoting cues to TAMs that further enhance tumorigenesis. Studies in the last decade have aimed to understand the molecular mediators of CSCs and TAMs, and recent advances have begun to elucidate the complex cross talk that occurs between these two cell types. In this review, we discuss the molecular interactions that define CSC-TAM cross talk at each stage of tumor progression and examine the clinical implications of targeting these interactions.

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Metastasis of Cancer Stem Cells Developed in the Microenvironment of Hepatocellular Carcinoma

Bioengineering ◽

10.3390/bioengineering6030073 ◽

2019 ◽

Vol 6 (3) ◽

pp. 73 ◽

Cited By ~ 7

Author(s):

Said M. Afify ◽

Ghmkin Hassan ◽

Amira Osman ◽

Anna Sanchez Calle ◽

Hend M Nawara ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Stem Cells ◽

Cancer Stem Cells ◽

Cultured Cells ◽

Malignant Tumors ◽

Metastatic Potential ◽

Metastatic Cells ◽

Huh7 Cells ◽

Induced Pluripotent

Metastasis develops when cancer cells spread from the primary site of a malignant tumor to the surrounding and distant tissues, and it is the most critical problem in cancer treatment. Our group developed cancer stem cells (CSCs) from induced pluripotent stem cells (iPSCs) in the presence of a conditioned medium (CM) of cancer-derived cells. The CSCs were characterized by the formation of malignant tumors in vivo, followed by metastasis. In this study, CSCs converted from mouse iPSCs in the presence of CM from hepatocellular carcinoma (HCC) cell line Huh7 cells. These converted cells (miPS-Huh7cm cells) were established as the metastatic cells. The generated CSCs were injected into the liver or spleen of nude mice. Almost one month after transplantation, the tumors were excised, and the primary cultured cells derived from the malignant tumors and metastatic nodules were evaluated by stemness and metastatic markers to compare their differences. The miPS-Huh7cm cells exhibited metastatic potential, and efficiently formed malignant tumors with lung and/or liver lesions in vivo, whereas the injected miPS formed teratoma. The primary cultured cells derived from the malignant tumors and metastatic nodules sustained the expression of stemness markers, such as Nanog, Klf4 and c-Myc, and acquired cancer stem markers, such as CD90, CD44 and ALDH1. Simultaneously, the expression of metastatic markers, such as Slug, Twist1 and vimentin, in primary cells derived from the malignant tumors, was higher than in metastatic nodules. The CSCs derived from iPSCs, forming malignant tumors and displaying high metastasis, will provide a good animal model to study the mechanisms of metastasis.

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Hypoxia, Inflammation and Necrosis as Determinants of Glioblastoma Cancer Stem Cells Progression

International Journal of Molecular Sciences ◽

10.3390/ijms21082660 ◽

2020 ◽

Vol 21 (8) ◽

pp. 2660

Author(s):

Marco Papale ◽

Mariachiara Buccarelli ◽

Cristiana Mollinari ◽

Matteo A. Russo ◽

Roberto Pallini ◽

...

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

Hypoxia Inducible Factor ◽

Human Tumor ◽

Metastatic Potential ◽

Cellular Adhesion ◽

Brain Invasion ◽

Hypoxia Inducible Factor 1 ◽

Tumor Tissues

Tumor hypoxic microenvironment causes hypoxia inducible factor 1 alpha (HIF-1α) activation and necrosis with alarmins release. Importantly, HIF-1α also controls the expression of alarmin receptors in tumor cells that can bind to and be activated by alarmins. Human tumor tissues possess 1–2% of cancer stem cells (CSCs) residing in hypoxic niches and responsible for the metastatic potential of tumors. Our hypothesis is that hypoxic CSCs express alarmin receptors that can bind alarmins released during necrosis, an event favoring CSCs migration. To investigate this aspect, glioblastoma stem-like cell (GSC) lines were kept under hypoxia to determine the expression of hypoxic markers as well as receptor for advanced glycation end products (RAGE). The presence of necrotic extracts increased migration, invasion and cellular adhesion. Importantly, HIF-1α inhibition by digoxin or acriflavine prevented the response of GSCs to hypoxia alone or plus necrotic extracts. In vivo, GSCs injected in one brain hemisphere of NOD/SCID mice were induced to migrate to the other one in which a necrotic extract was previously injected. In conclusion, our results show that hypoxia is important not only for GSCs maintenance but also for guiding their response to external necrosis. Inhibition of hypoxic pathway may therefore represent a target for preventing brain invasion by glioblastoma stem cells (GSCs).

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Combination of Histone Deacetylase Inhibitor with Cu(II) 5,5- diethylbarbiturate Complex Induces Apoptosis in Breast Cancer Stem Cells: A Promising Novel Approach

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520621666201207090702 ◽

2020 ◽

Vol 21 ◽

Author(s):

Merve Erkisa ◽

Nazlihan Aztopal ◽

Elif Erturk ◽

Engin Ulukaya ◽

Veysel T. Yilmaz ◽

...

Keyword(s):

Breast Cancer ◽

Oxidative Stress ◽

Stem Cells ◽

Cancer Stem Cells ◽

Histone Deacetylases ◽

Caspase 3 ◽

Cancer Cell Line ◽

Annexin V ◽

Tumor Formation ◽

Dependent Manner

Background: Cancer stem cells (CSC) are subpopulation within the tumor that acts a part in the initiation, progression, recurrence, resistance to drugs and metastasis of cancer. It is well known that epigenetic changes lead to tumor formation in cancer stem cells and show drug resistance. Epigenetic modulators and /or their combination with different agents have been used in cancer therapy. Objective: In our study we scope out the effects of combination of a histone deacetylases inhibitor, valproic acid (VPA), and Cu(II) complex [Cu(barb-κN)(barb-κ2N,O)(phen-κN,N’)]·H2O] on cytotoxicity/apoptosis in a stem-cell enriched population (MCF-7s) obtained from parental breast cancer cell line (MCF-7). Methods: Viability of the cells was measured by the ATP assay. Apoptosis was elucidated via the assessment of caspase-cleaved cytokeratin 18 (M30 ELISA) and a group of flow cytometry analysis (caspase 3/7 activity, phosphatidylserine translocation by annexin V-FITC assay, DNA damage and oxidative stress) and 2ˈ,7ˈ–dichlorofluorescein diacetate staining. Results: The VPA combined with Cu(II) complex showed anti proliferative activity on MCF-7s cells in a dose- and time-dependently. Treatment with combination of 2.5 mM VPA and 3.12 μM Cu(II) complex induces oxidative stress in a time-dependent manner, as well as apoptosis that is evidenced by the increase in caspase 3/7 activity, positive annexin-V-FITC, and increase in M30 levels. Conclusion: The results suggest that the combination therapy induces apoptosis following increased oxidative stress, thereby making it a possible promising therapeutic strategy that further analysis is required.

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STEM-26. SMALL MOLECULE DRUG CBL0137 INHIBITS THE FACT COMPLEX AND SENSITIZES GLIOBLASTOMA CANCER STEM CELLS TO RADIOTHERAPY

Neuro-Oncology ◽

10.1093/neuonc/noaa215.843 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii201-ii202

Author(s):

Miranda Tallman ◽

Abigail Zalenski ◽

Amanda Deighen ◽

Morgan Schrock ◽

Sherry Mortach ◽

...

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

High Rate ◽

Orthotopic Model ◽

In Vivo Models ◽

Specific Cytotoxicity ◽

Treatment Paradigm ◽

Cancer Agent

Abstract Glioblastoma (GBM) is a malignant brain tumor with nearly universal recurrence. GBM cancer stem cells (CSCs), a subpopulation of radio- and chemo-resistant cancer cells capable of self-renewal, contribute to the high rate of recurrence. The anti-cancer agent, CBL0137, inhibits the FACT (facilitates chromatin transcription) complex leading to cancer cell specific cytotoxicity. Here, we show that CBL0137 sensitized GBM CSCs to radiotherapy using both in vitro and in vivo models. Treatment of CBL0137 combined with radiotherapy led to increased DNA damage in GBM patient specimens and failure to resolve the damage led to decreased cell viability. Using clonogenic assays, we confirmed that CBL0137 radiosensitized the CSCs. To validate that combination therapy impacted CSCs, we used an in vivo subcutaneous model and showed a decrease in the frequency of cancer stem cells present in tumors as well as decreased tumor volume. Using an orthotopic model of GBM, we confirmed that treatment with CBL0137 followed by radiotherapy led to significantly increased survival compared to either treatment alone. Radiotherapy remains a critical component of patient care for GBM, even though there exists a resistant subpopulation. Radio-sensitizing agents, including CBL0137, pose an exciting treatment paradigm to increase the efficacy of irradiation, especially by inclusively targeting CSCs.

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STEM-20. A CANCER STEM CELL-SELECTIVE APOPTOSIS-INDUCING SMALL MOLECULE FOR THE TREATMENT OF GBM

Neuro-Oncology ◽

10.1093/neuonc/noaa215.837 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii200-ii200

Author(s):

Stephen Skirboll ◽

Natasha Lucki ◽

Genaro Villa ◽

Naja Vergani ◽

Michael Bollong ◽

...

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

Small Molecules ◽

Small Molecule ◽

Large Scale ◽

Critical Role ◽

Tumor Formation ◽

Cell Type ◽

Caspase 1 ◽

Activation Assay

Abstract INTRODUCTION Glioblastoma multiforme (GBM) is the most aggressive form of primary brain cancer. A subpopulation of multipotent cells termed GBM cancer stem cells (CSCs) play a critical role in tumor initiation and maintenance, drug resistance, and recurrence following surgery. New therapeutic strategies for the treatment of GBM have recently focused on targeting CSCs. Here we have used an unbiased large-scale screening approach to identify drug-like small molecules that induce apoptosis in GBM CSCs in a cell type-selective manner. METHODS A luciferase-based survival assay of patient-derived GBM CSC lines was established to perform a large-scale screen of ∼one million drug-like small molecules with the goal of identifying novel compounds that are selectively toxic to chemoresistant GBM CSCs. Compounds found to kill GBM CSC lines as compared to control cell types were further characterized. A caspase activation assay was used to evaluate the mechanism of induced cell death. A xenograft animal model using patient-derived GBM CSCs was employed to test the leading candidate for suppression of in vivo tumor formation. RESULTS We identified a small molecule, termed RIPGBM, from the cell-based chemical screen that induces apoptosis in primary patient-derived GBM CSC cultures. The cell type-dependent selectivity of RIPGBM appears to arise at least in part from redox-dependent formation of a proapoptotic derivative, termed cRIPGBM, in GBM CSCs. cRIPGBM induces caspase 1-dependent apoptosis by binding to receptor-interacting protein kinase 2 (RIPK2) and acting as a molecular switch, which reduces the formation of a prosurvival RIPK2/TAK1 complex and increases the formation of a proapoptotic RIPK2/caspase 1 complex. In an intracranial GBM xenograft mouse model, RIPGBM was found to significantly suppress tumor formation. CONCLUSIONS Our chemical genetics-based approach has identified a small molecule drug candidate and a potential drug target that selectively targets cancer stem cells and provides an approach for the treatment of GBMs.

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STEM-20. RADIO-SENSITIZING GLIOBLASTOMA CANCER STEM CELLS BY INHIBITION OF FACT

Neuro-Oncology ◽

10.1093/neuonc/noz175.993 ◽

2019 ◽

Vol 21 (Supplement_6) ◽

pp. vi237-vi238

Author(s):

Miranda Montgomery ◽

Abigail Zalenski ◽

Amanda Deighen ◽

Sherry Mortach ◽

Treg Grubb ◽

...

Keyword(s):

Stem Cells ◽

Dna Damage ◽

Cancer Stem Cells ◽

Combination Therapy ◽

High Rate ◽

Primary Role ◽

Cancer Cell Growth ◽

Treatment Paradigm

Abstract Glioblastoma (GBM) has a particularly high rate of recurrence with a 5-year overall survival rate of approximately 5%. This is in part due to a sub-population of cancer stem cells (CSC), which are both radioresistant and chemotherapeutically resistant to conventional treatments. Here we investigated CBL0137, a small molecule form of curaxin, in combination with radiotherapy as a means to radiosensitize CSCs. CBL0137 sequesters FACT (facilitates chromatin transcription) complex to chromatin, which leads to activation of p53 and inhibition of NF-κB. This sequestering of FACT results in cytotoxicity especially within tumor cells and prevents FACT from performing its primary role as a histone chaperone, as well as inhibits its part in the DNA damage response pathway. We show that when combined with radiotherapy, CBL0137 administration limited the ability of CSCs to identify and repair damaged DNA. CSCs treated in vitro with CBL0137 and irradiation showed an increased inhibition of cancer cell growth and decreased viability compared to irradiation or drug alone. Combination therapy also showed more DNA damage in the CSCs than with either agent alone. Based on our in vitro evidence for the efficacy of combination therapy to target CSCs, we moved forward to test the treatment in vivo. Using a subcutaneous model, we show that the amount of CD133+ cells (a marker for GMB CSCs) was reduced in irradiation plus CBL0137 compared to either treatment alone. Survival studies demonstrated that irradiation plus CBL0137 compared to irradiation alone or CBL0137 alone increase lifespan. Here we show the ability of CBL0137, in combination with irradiation, to target patient GBM CSCs both in vitro and in vivo. This work establishes a new treatment paradigm for GBM that inclusively targets CSCs and may ultimately reduce tumor recurrence.

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Targeting specificity of dendritic cells on breast cancer stem cells: in vitro and in vivo evaluations

OncoTargets and Therapy ◽

10.2147/ott.s77554 ◽

2015 ◽

pp. 323 ◽

Cited By ~ 3

Author(s):

Phuc Pham ◽

Sinh Nguyen ◽

Viet Pham ◽

Ngoc Phan ◽

Huyen Nguyen ◽

...

Keyword(s):

Breast Cancer ◽

Stem Cells ◽

Dendritic Cells ◽

Cancer Stem Cells ◽

Breast Cancer Stem Cells

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Genome-wide CRISPR-Cas9 screen identified KLF11 as a druggable suppressor for sarcoma cancer stem cells

Science Advances ◽

10.1126/sciadv.abe3445 ◽

2021 ◽

Vol 7 (5) ◽

pp. eabe3445

Author(s):

Yicun Wang ◽

Jinhui Wu ◽

Hui Chen ◽

Yang Yang ◽

Chengwu Xiao ◽

...

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

Negative Feedback ◽

Direct Interaction ◽

Therapy Resistance ◽

Negative Regulator ◽

Chemotherapy Response ◽

Genome Wide

Cancer stem cells (CSCs) are involved in tumorigenesis, recurrence, and therapy resistance. To identify critical regulators of sarcoma CSCs, we performed a reporter-based genome-wide CRISPR-Cas9 screen and uncovered Kruppel-like factor 11 (KLF11) as top candidate. In vitro and in vivo functional annotation defined a negative role of KLF11 in CSCs. Mechanistically, KLF11 and YAP/TEAD bound to adjacent DNA sites along with direct interaction. KLF11 recruited SIN3A/HDAC to suppress the transcriptional output of YAP/TEAD, which, in turn, promoted KLF11 transcription, forming a negative feedback loop. However, in CSCs, this negative feedback was lost because of epigenetic silence of KLF11, causing sustained YAP activation. Low KLF11 was associated with poor prognosis and chemotherapy response in patients with sarcoma. Pharmacological activation of KLF11 by thiazolidinedione effectively restored chemotherapy response. Collectively, our study identifies KLF11 as a negative regulator in sarcoma CSCs and potential therapeutic target.

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