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

SLAS Discovery
Victor QueredaDerek Duckett

Abstract

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).

References

Nov 19, 2004·Nature·Sheila K SinghPeter B Dirks
Jan 28, 2005·Journal of Neuro-oncology·Tim Demuth, Michael E Berens
Nov 3, 2007·Genes & Development·Frank B FurnariWebster K Cavenee
Feb 9, 2008·Molecular and Cellular Biochemistry·Chen Lu, Amal Shervington
Sep 16, 2008·Bioconjugate Chemistry·Thomas Tyrel GoodmanSuzie Hwang Pun
Dec 8, 2010·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Albert LaiTimothy Cloughesy
Mar 26, 2013·Biochemical and Biophysical Research Communications·Xiong JinHyunggee Kim
Jun 22, 2013·Bioorganic & Medicinal Chemistry Letters·Mathieu BibianWilliam R Roush
Jul 24, 2014·Cancer Epidemiology, Biomarkers & Prevention : a Publication of the American Association for Cancer Research, Cosponsored by the American Society of Preventive Oncology·Jigisha P ThakkarJohn L Villano
Aug 15, 2014·Biotechnology Advances·Xian XuXinqiao Jia
Mar 26, 2015·Anatomy & Cell Biology·Maryam RahmanBrent A Reynolds
Jun 26, 2015·Genes & Development·Justin D LathiaJeremy N Rich
Dec 17, 2015·JAMA : the Journal of the American Medical Association·Roger StuppZvi Ram
Feb 16, 2016·Expert Opinion on Therapeutic Targets·Laura Garros-RegulezAnder Matheu
Dec 21, 2016·Current Cancer Drug Targets·Drew A SpencerMaciej S Lesniak
Nov 1, 2011·Australasian Journal of Ultrasound in Medicine·Cindy Oakenful, Andreas L Lambrianides

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Citations

Mar 16, 2019·Annals of the New York Academy of Sciences·M Catarina Silva, Stephen J Haggarty
Aug 30, 2020·Nanomaterials·Kyle Bromma, Devika B Chithrani
Jun 23, 2019·Scientific Reports·BanuPriya SridharanTimothy P Spicer
Mar 7, 2021·Cancers·Sreenivasulu GuntiNyall R London

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Methods Mentioned

BETA
FCS
Assay
fluorescence microscopy
confocal microscopy

Software Mentioned

GraphPad Prism
Assay Explorer
Scripps
ImageJ

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