scholarly journals Large-Scale Drug Screening in Patient-Derived IDHmut Glioma Stem Cells Identifies Several Efficient Drugs among FDA-Approved Antineoplastic Agents

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1389 ◽  
Author(s):  
Philip Dao Trong ◽  
Gerhard Jungwirth ◽  
Tao Yu ◽  
Stefan Pusch ◽  
Andreas Unterberg ◽  
...  
Keyword(s):  
Stem Cells ◽  
Drug Screening ◽  
Large Scale ◽  
Annexin V ◽  
Chemotherapeutic Agents ◽  
Glioma Stem Cells ◽  
Clinical Investigations ◽  
Ic50 Values ◽  
Induction Of Apoptosis ◽  
Approved Drugs

The discovery of the isocitrate dehydrogenase (IDH) mutation in glioma led to a paradigm shift on how we see glioma biology. Difficulties in cultivating IDH mutant glioma stem cells (IDHmut GSCs) resulted in a paucity of preclinical models in IDHmut glioma, limiting the discovery of new effective chemotherapeutic agents. To fill this gap, we used six recently developed patient-derived IDHmut GSC lines and performed a large-scale drug screening with 147 Food and Drug Administration (FDA)-approved anticancer drugs. GSCs were subjected to the test compounds for 72 h in concentrations ranging from 0.0001 to 1 µM. Cell viability was assessed by CellTiterGlo and the induction of apoptosis by flow cytometry with Annexin V/propidium iodide staining. The initial screen was performed with two IDHmut GSC lines and identified seven drugs (bortezomib, carfilzomib, daunorubicin, doxorubicin, epirubicin, omacetaxine, plicamycin) with a substantial antiproliferative activity, as reflected by half maximal inhibitory concentrations (IC50) below 1 µM and maximum inhibitory effects (Emax) below 25%. These findings were validated in an additional four IDHmut GSC lines. The candidate drugs, of which plicamycin and omacetaxine are known to cross the blood brain barrier, were used for subsequent cell death analyses. A significant induction of apoptosis was observed at IC50 values of the respective drugs. In summary, we were able to identify seven FDA-approved drugs that should be further taken into clinical investigations for the treatment of IDHmut gliomas.

scholarly journals DDIS-22. DRUG SCREEN IN PATIENT-DERIVED IDHMUT GLIOMA STEM CELLS IDENTIFIES SEVERAL FDA-APPROVED ANTINEOPLASTIC AGENTS

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi67-vi68
Author(s):  
Philip Dao Trong ◽  
Stefan Pusch ◽  
Andreas Unterberg ◽  
Christel Herold-Mende ◽  
Rolf Warta
Keyword(s):  
Stem Cells ◽  
Antineoplastic Agents ◽  
Chemotherapeutic Agents ◽  
Cell Culture Supernatant ◽  
Drug Screen ◽  
Preclinical Model ◽  
Glioma Stem Cells ◽  
Neurosurgical Department ◽  
Stable Production ◽  
Approved Drugs

Abstract OBJECTIVE The discovery of the Isocitrate Dehydrogenase (IDH) mutation in glioma has led to a paradigm shift on how we see glioma biology. While it is clear, that IDH mutated (IDHmut) and wildtype (IDHwt) tumors have to be viewed as separate entities, the underlying biological differences are still matter of extensive research. Difficulties in cultivating IDHmut glioma stem cells (GSC) have led to a paucity of preclinical models in IDHmut glioma making the discovery of new effective chemotherapeutic agents problematic. We therefore sought to perform a repurposing drug screen in five patient-derived IDHmut GSC lines to discover potential effective antineoplastic agents, already approved by the FDA. METHODS Patient tumor tissue was obtained in our neurosurgical department to isolate and establish IDHmut GSC lines. (D)-2-hydroxyglutarate (2HG) levels were measured in the cell culture supernatant of IDHmut GSCs using an enzymatic diaphorase/resazurin system. The drug library provided by the Developmental Therapeutics Program (DTP) of the National Cancer Institute (NCI) consisting of 146 FDA-approved drugs was used for the screen. Cell viability was assessed with the CellTiterGlo assay (Promega). RESULTS Despite several passages, the IDHmut GSCs showed stable production of 2HG and were therefore suitable for the drug screen. Cells were cultured as neurospheres and subjected to the test compounds for 72h in concentrations ranging from 0.1nM – 1µM. We identified several compounds in two IDHmut GSC lines (NCH551b, NCH1681) that had a half maximal inhibitory concentration (IC50) below 1µM and could confirm its cytotoxic potential in additional three IDHmut GSC lines (NCH612, NCH620, NCH3763). CONCLUSION In this study, we present a feasible preclinical model for a high-throughput drug screen in patient-derived IDHmut GSCs and identified several FDA-approved antineoplastic agents which warrant further investigations.

scholarly journals Natural Products Targeting Cancer Stem Cells for Augmenting Cancer Therapeutics

2021 ◽  
Vol 22 (23) ◽  
pp. 13044
Author(s):  
Ari Meerson ◽  
Soliman Khatib ◽  
Jamal Mahajna
Keyword(s):  
Stem Cells ◽  
Natural Products ◽  
Cancer Stem Cells ◽  
Signaling Pathways ◽  
Tumor Cells ◽  
Cancer Therapeutics ◽  
Chemotherapeutic Agents ◽  
Biologically Active ◽  
Approved Drugs ◽  
Multiple Signaling

Cancer stem cells (CSC) have been identified in several types of solid tumors. In some cases, CSC may be the source of all the tumor cells, the cause of the tumor’s resistance to chemotherapeutic agents, and the source of metastatic cells. Thus, a combination therapy targeting non-CSC tumor cells as well as specifically targeting CSCs holds the potential to be highly effective. Natural products (NPs) have been a historically rich source of biologically active compounds and are known for their ability to influence multiple signaling pathways simultaneously with negligible side effects. In this review, we discuss the potential of NPs in targeting multiple signaling pathways in CSC and their potential to augment the efficacy of standard cancer therapy. Specifically, we focus on the anti-CSC activities of flavonoids, FDA-approved drugs originating from natural sources. Additionally, we emphasize the potential of NPs in targeting microRNA-mediated signaling, given the roles of microRNA in the maintenance of the CSC phenotype.

A Novel Peptide Containing a Nine Amino Acid Sequence from Nur77 Induces Bcl-2-Dependent Apoptosis in AML.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2607-2607
Author(s):  
Michael Andreeff ◽  
Marina Konopleva ◽  
Julie C. Watt ◽  
Ismael J. Samudio ◽  
Arnold C. Satterthwait ◽  
...  
Keyword(s):  
Stem Cells ◽  
Amino Acid ◽  
Conformational Change ◽  
Annexin V ◽  
Orphan Receptor ◽  
Cytochrome C Release ◽  
Potent Inducer ◽  
Amino Acid Peptide ◽  
Induction Of Apoptosis ◽  
Hct116 Cells

Abstract Nur77 (also known as TR3 and NGFI-B) is a nuclear orphan receptor that in response to various stimuli can translocate from the nucleus to the mitochondria, bind Bcl-2, and induce a conformational change in Bcl-2 that exposes its BH3 domain. This conformational change of Bcl-2 transforms this protein into a pro-apoptotic molecule that can induce cytochrome c release and apoptosis (Cell 116:527, 2004). Interestingly, in acute myeloid leukemia (AML) cell lines and blasts, Nur77 is absent. We have recently generated a nine amino acid peptide from the Nur77 protein (TR3) that is capable of inducing the same pro-apoptotic conformational change in Bcl-2 that Nur77 elicits (manuscript submitted). We tested the hypothesis that a cell-permeable version of this peptide can induce Bcl-2-dependent apoptosis in AML cells and AML stem cells. Primary AML samples (n=6) were incubated with 20 μM control or TR3 peptide for 24 hrs. Induction of apoptosis was measured as CD34+38−123+ PS/Annexin V+ by multiparametric flow cytometry. Our results demonstrate that the L-enantiomer form of TR3 indeed induces apoptosis in total AML cells (60%, p=0.001) and CD34+38−123+ stem cells (67%, p=0.001), as compared to control peptide. Bax expression did not affect the peptide’s ability to induce apoptosis, as wild-type HCT116 cells and HCT116 cells deficient in Bax showed similar levels of apoptosis after treatment with the TR3 peptide. However, Bcl-2 expression was shown to be critical since lentiviral shRNA ablation of Bcl-2 in KG1 cells completely prevented induction of apoptosis by TR3 peptides. We conclude that the TR3 peptide is a potent inducer of apoptosis that mimics the action of Nur77 on Bcl-2 at the mitochondrial level. These results suggest that the Nur77 peptide or compounds that mimic this peptide may have utility as a novel therapeutic agent against Bcl-2 expressing cancers and leukemias.

scholarly journals The Effect of Furanocoumarin Derivatives on Induction of Apoptosis and Multidrug Resistance in Human Leukemic Cells

Molecules ◽  
2019 ◽  
Vol 24 (9) ◽  
pp. 1824 ◽  
Author(s):  
Tomasz Kubrak ◽  
Marcin Czop ◽  
Przemysław Kołodziej ◽  
Marta Ziaja-Sołtys ◽  
Jacek Bogucki ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Cell Lines ◽  
Leukemia Cell ◽  
Annexin V ◽  
Leukemic Cells ◽  
Leukemia Cells ◽  
Human Leukemia ◽  
Ic50 Values ◽  
Induction Of Apoptosis ◽  
Presence And Absence

Background: The insensitivity of cancer cells to therapeutic agents is considered to be the main cause of failure of therapy and mortality of patients with cancer. A particularly important problem in these patients is the phenomenon of multidrug resistance, consisting of abnormal, elevated expression of transport proteins (ABC family). The aim of this research included determination of IC50 values of selected furanocoumarins in the presence and absence of mitoxantrone in leukemia cells and analysis of changes in apoptosis using anexinV/IP and Casp3/IP after 24 h exposure of cell lines to selected coumarins in the presence and absence of mitoxantrone in IC50 concentrations. Methods: Research was conducted on 3 cell lines derived from the human hematopoietic system: HL-60, HL-60/MX1 and HL-60/MX2. After exposure to coumarin compounds, cells were subjected to cytometric analysis to determine the induction of apoptosis by two methods: the Annexin V test with propidium iodide and the PhiPhiLux-G1D2 reagent containing caspase 3 antibodies. Results: All of the furanocoumarin derivatives studied were found to induce apoptosis in leukemia cell lines. Conclusions: Our results clearly show that the furanocoumarin derivatives are therapeutic substances with antitumor activity inducing apoptosis in human leukemia cells with phenotypes of resistance.

scholarly journals A novel artificial lung organoid for simulating a patient derived adenocarcinoma of lung for personalized oncology

2021 ◽  
Author(s):  
Sally Esmail ◽  
Wayne R Danter
Keyword(s):  
Drug Screening ◽  
Large Scale ◽  
Disease Modeling ◽  
Standard Of Care ◽  
Cancer Drug ◽  
Genetic Profile ◽  
Precision Oncology ◽  
Mutational Profiling ◽  
Approved Drugs ◽  
The Individual

ABSTRACTOptimizing patient care based on precision oncology will inevitably become the standard of care. If we accept the principle that every persons’ cancer is different then the most effective therapies will have to be designed for the individual patient and for their tumors genetic profile. Access to tumor mutational profiling is now widely available but continues to be limited by cost and actionable information. For example, novel combinations of approved drugs are rarely considered. These considerations lead us to hypothesize that artificially induced Lung Adenocarcinoma (LUAD) derived lung organoids could provide a novel, alternate approach for LUAD disease modeling and large-scale targeted drug screening.In this project, we used data from a commercially available tumor mutation profile to generate and then validate the artificially induced LUAD-derived lung organoid simulations (aiLUNG-LUAD) to model LUAD and identify several drug combinations that effectively reverse the tumors’ genotypic and phenotypic features when compared with placebo. These results complement previous LUAD-derived lung organoids research and provide a novel and widely applicable cancer drug-screening approach for precision/individualized oncology.

scholarly journals A Cytotoxic Three-Dimensional-Spheroid, High-Throughput Assay Using Patient-Derived Glioma Stem Cells

2018 ◽  
Vol 23 (8) ◽  
pp. 842-849 ◽  
Author(s):  
Victor Quereda ◽  
Shurong Hou ◽  
Franck Madoux ◽  
Louis Scampavia ◽  
Timothy P. Spicer ◽  
...  
Keyword(s):  
Stem Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Large Scale ◽  
Three Dimensional ◽  
Treatment Resistance ◽  
Small Population ◽  
Molecular Probes ◽  
Glioma Stem Cells

Glioblastoma (GBM) is the most aggressive primary brain cancer with an average survival time after diagnosis of only 12–14 months, with few (<5%) long-term survivors. A growing body of work suggests that GBMs contain a small population of glioma stem cells (GSCs) that are thought to be major contributors to treatment resistance and disease relapse. Identifying compounds that modulate GSC proliferation would provide highly valuable molecular probes of GSC-directed signaling. However, targeting GSCs pharmacologically has been challenging. Patient-derived GSCs can be cultured as neurospheres, and in vivo these cells functionally recapitulate the heterogeneity of the original tumor. Using patient-derived GSC-enriched cultures, we have developed a 1536-well spheroid-based proliferation assay and completed a pilot screen, testing ~3300 compounds comprising approved drugs. This cytotoxic and automation-friendly assay yielded a signal-to-background (S/B) ratio of 161.3 ± 7.5 and Z′ of 0.77 ± 0.02, demonstrating its robustness. Importantly, compounds were identified with anti-GSC activity, demonstrating the applicability of this assay for large-scale high-throughput screening (HTS).

scholarly journals EXTH-14. REPURPOSING OF APPROVED DRUGS FOR THE TREATMENT OF GLIOMA STEM CELLS

2018 ◽  
Vol 20 (suppl_6) ◽  
pp. vi87-vi88
Author(s):  
Sang Lee ◽  
Becky Slagle-Webb ◽  
Brad Zacharia ◽  
James Connor
Keyword(s):  
Stem Cells ◽  
Glioma Stem Cells ◽  
Approved Drugs

scholarly journals Large Scale Identification of Variant Proteins in Glioma Stem Cells

2017 ◽  
Vol 9 (1) ◽  
pp. 73-79 ◽  
Author(s):  
Ekaterina Mostovenko ◽  
Ákos Végvári ◽  
Melinda Rezeli ◽  
Cheryl F. Lichti ◽  
David Fenyö ◽  
...  
Keyword(s):  
Stem Cells ◽  
Large Scale ◽  
Glioma Stem Cells

The Effect of Active Site-inhibited Factor VIIa on Tissue Factor-initiated Coagulation Using Platelets before and after Aspirin Administration

1997 ◽  
Vol 78 (04) ◽  
pp. 1202-1208 ◽  
Author(s):  
Marianne Kjalke ◽  
Julie A Oliver ◽  
Dougald M Monroe ◽  
Maureane Hoffman ◽  
Mirella Ezban ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Active Site ◽  
Large Scale ◽  
Thrombin Generation ◽  
Factor Viia ◽  
Dependent Manner ◽  
Antithrombotic Agent ◽  
Before And After ◽  
Ic50 Values

SummaryActive site-inactivated factor VIIa has potential as an antithrombotic agent. The effects of D-Phe-L-Phe-L-Arg-chloromethyl ketone-treated factor VIla (FFR-FVIIa) were evaluated in a cell-based system mimicking in vivo initiation of coagulation. FFR-FVIIa inhibited platelet activation (as measured by expression of P-selectin) and subsequent large-scale thrombin generation in a dose-dependent manner with IC50 values of 1.4 ± 0.8 nM (n = 8) and 0.9 ± 0.7 nM (n = 7), respectively. Kd for factor VIIa binding to monocytes ki for FFR-FVIIa competing with factor VIIa were similar (11.4 ± 0.8 pM and 10.6 ± 1.1 pM, respectively), showing that FFR-FVIIa binds to tissue factor in the tenase complex with the same affinity as factor VIIa. Using platelets from volunteers before and after ingestion of aspirin (1.3 g), there were no significant differences in the IC50 values of FFR-FVIIa [after aspirin ingestion, the IC50 values were 1.7 ± 0.9 nM (n = 8) for P-selectin expression, p = 0.37, and 1.4 ± 1.3 nM (n = 7) for thrombin generation, p = 0.38]. This shows that aspirin treatment of platelets does not influence the inhibition of tissue factor-initiated coagulation by FFR-FVIIa, probably because thrombin activation of platelets is not entirely dependent upon expression of thromboxane A2.

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