Skip to main content

Identification of Novel Molecules Targeting Cancer Stem Cells

  • Protocol
  • First Online:
Colorectal Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1765))

Abstract

Multiple studies focused on tumor heterogeneity and cellular hierarchies have demonstrated the role of cancer stem cells (CSC) in tumor initiation and recurrence. Colorectal cancer is one of the leading causes of cancer-related death and is hierarchically organized, with the majority of tumor cells descending from a small population of colon cancer stem cells (CCSCs). Such a rare self-renewing population is marked by the acquisition of distinct chromatin regulation and transcriptional programs. Fundamental molecular deviations between CCSCs and bulk tumor cells as well as normal tissues represent a unique therapeutic access to develop novel, selective anticancer therapies.

In this chapter, we describe a methodological pipeline to identify novel molecules to selectively target human CCSC. We present a point-by-point description of a typical phenotypic molecular screening experiment, aiming to identify selective modulators of human CCSCs vs. normal intestinal progenitor cells.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Bonnet D, Dick JE (1997) Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med 3(7):730–737

    Article  CAS  PubMed  Google Scholar 

  2. Visvader JE, Lindeman GJ (2012) Cancer stem cells: current status and evolving complexities. Cell Stem Cell 10(6):717–728. https://doi.org/10.1016/j.stem.2012.05.007

    Article  CAS  PubMed  Google Scholar 

  3. Kreso A, Dick JE (2014) Evolution of the cancer stem cell model. Cell Stem Cell 14(3):275–291. https://doi.org/10.1016/j.stem.2014.02.006

    Article  CAS  PubMed  Google Scholar 

  4. Siegel RL, Miller KD, Jemal A (2016) Cancer statistics, 2016. CA Cancer J Clin 66(1):7–30. https://doi.org/10.3322/caac.21332

    Article  PubMed  Google Scholar 

  5. O'Brien CA, Pollett A, Gallinger S, Dick JE (2007) A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445(7123):106–110. https://doi.org/10.1038/nature05372

    Article  PubMed  Google Scholar 

  6. Widschwendter M, Fiegl H, Egle D, Mueller-Holzner E, Spizzo G, Marth C, Weisenberger DJ, Campan M, Young J, Jacobs I, Laird PW (2007) Epigenetic stem cell signature in cancer. Nat Genet 39(2):157–158. https://doi.org/10.1038/ng1941

    Article  CAS  PubMed  Google Scholar 

  7. Suva ML, Riggi N, Bernstein BE (2013) Epigenetic reprogramming in cancer. Science 339(6127):1567–1570. https://doi.org/10.1126/science.1230184

    Article  CAS  PubMed  Google Scholar 

  8. Merlos-Suárez A, Barriga FM, Jung P, Iglesias M, Céspedes MV, Rossell D, Sevillano M, Hernando-Momblona X, da Silva-Diz V, Muñoz P, Clevers H, Sancho E, Mangues R, Batlle E (2011) The intestinal stem cell signature identifies colorectal cancer stem cells and predicts disease relapse. Cell Stem Cell 8(5):511–524. https://doi.org/10.1016/j.stem.2011.02.020

    Article  PubMed  Google Scholar 

  9. Williams R (2013) Discontinued drugs in 2012: oncology drugs. Expert Opin Investig Drugs 22(12):1627–1644. https://doi.org/10.1517/13543784.2013.847088

    Article  CAS  PubMed  Google Scholar 

  10. Kotz J (2012) Phenotypic screening, take two. Sci BX 5(15). https://doi.org/10.1038/scibx.2012.380

  11. Swinney DC (2013) Phenotypic vs. target-based drug discovery for first-in-class medicines. Clin Pharmacol Ther 93(4):299–301. https://doi.org/10.1038/clpt.2012.236

    Article  CAS  PubMed  Google Scholar 

  12. Swinney DC, Anthony J (2011) How were new medicines discovered? Nat Rev Drug Discov 10(7):507–519. https://doi.org/10.1038/nrd3480

    Article  CAS  PubMed  Google Scholar 

  13. Benoit YD, Guezguez B, Boyd AL, Bhatia M (2014) Molecular pathways: epigenetic modulation of Wnt-glycogen synthase kinase-3 signaling to target human cancer stem cells. Clin Cancer Res 20(21):5372–5378. https://doi.org/10.1158/1078-0432.CCR-13-2491

    Article  CAS  PubMed  Google Scholar 

  14. Kahn M (2014) Can we safely target the WNT pathway? Nat Rev Drug Discov 13(7):513–532. https://doi.org/10.1038/nrd4233

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Sachlos E, Risueño RM, Laronde S, Shapovalova Z, Lee JH, Russell J, Malig M, McNicol JD, Fiebig-Comyn A, Graham M, Levadoux-Martin M, Lee JB, Giacomelli AO, Hassell JA, Fischer-Russell D, Trus MR, Foley R, Leber B, Xenocostas A, Brown ED, Collins TJ, Bhatia M (2012) Identification of drugs including a dopamine receptor antagonist that selectively target cancer stem cells. Cell 149(6):1284–1297. https://doi.org/10.1016/j.cell.2012.03.049

    Article  CAS  PubMed  Google Scholar 

  16. Hughes JP, Rees S, Kalindjian SB, Philpott KL (2011) Principles of early drug discovery. Br J Pharmacol 162(6):1239–1249. https://doi.org/10.1111/j.1476-5381.2010.01127.x

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Zhang JH, Chung TD, Oldenburg KR (1999) A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 4(2):67–73. https://doi.org/10.1177/108705719900400206

    Article  CAS  PubMed  Google Scholar 

  18. Thorne N, Auld DS, Inglese J (2010) Apparent activity in high-throughput screening: origins of compound-dependent assay interference. Curr Opin Chem Biol 14(3):315–324. https://doi.org/10.1016/j.cbpa.2010.03.020

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  19. Gupta PB, Onder TT, Jiang G, Tao K, Kuperwasser C, Weinberg RA, Lander ES (2009) Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell 138(4):645–659. https://doi.org/10.1016/j.cell.2009.06.034

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  20. Yasgar A, Titus SA, Wang Y, Danchik C, Yang SM, Vasiliou V, Jadhav A, Maloney DJ, Simeonov A, Martinez NJ (2017) A high-content assay enables the automated screening and identification of small molecules with specific ALDH1A1-inhibitory activity. PLoS One 12(1):e0170937. https://doi.org/10.1371/journal.pone.0170937

    Article  PubMed Central  PubMed  Google Scholar 

  21. Perreault N, Beaulieu JF (1996) Use of the dissociating enzyme thermolysin to generate viable human normal intestinal epithelial cell cultures. Exp Cell Res 224(2):354–364

    Article  CAS  PubMed  Google Scholar 

  22. Prasetyanti PR, Zimberlin C, De Sousa E, Melo F, Medema JP (2013) Isolation and propagation of colon cancer stem cells. Methods Mol Biol 1035:247–259. https://doi.org/10.1007/978-1-62703-508-8_21

    Article  CAS  PubMed  Google Scholar 

  23. Benoit YD, Lussier C, Ducharme PA, Sivret S, Schnapp LM, Basora N, Beaulieu JF (2009) Integrin alpha8beta1 regulates adhesion, migration and proliferation of human intestinal crypt cells via a predominant RhoA/ROCK-dependent mechanism. Biol Cell 101(12):695–708. https://doi.org/10.1042/BC20090060

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Benoit YD, Pare F, Francoeur C, Jean D, Tremblay E, Boudreau F, Escaffit F, Beaulieu JF (2010) Cooperation between HNF-1alpha, Cdx2, and GATA-4 in initiating an enterocytic differentiation program in a normal human intestinal epithelial progenitor cell line. Am J Physiol Gastrointest Liver Physiol 298(4):G504–G517. https://doi.org/10.1152/ajpgi.00265.2009

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  25. Sikandar SS, Pate KT, Anderson S, Dizon D, Edwards RA, Waterman ML, Lipkin SM (2010) NOTCH signaling is required for formation and self-renewal of tumor-initiating cells and for repression of secretory cell differentiation in colon cancer. Cancer Res 70(4):1469–1478. https://doi.org/10.1158/0008-5472.CAN-09-2557

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  26. Barbaric I, Jones M, Harley DJ, Gokhale PJ, Andrews PW (2011) High-content screening for chemical modulators of embryonal carcinoma cell differentiation and survival. J Biomol Screen 16(6):603–617. https://doi.org/10.1177/1087057111406547

    Article  CAS  PubMed  Google Scholar 

  27. Vermeulen L, De Sousa E, Melo F, van der Heijden M, Cameron K, de Jong JH, Borovski T, Tuynman JB, Todaro M, Merz C, Rodermond H, Sprick MR, Kemper K, Richel DJ, Stassi G, Medema JP (2010) Wnt activity defines colon cancer stem cells and is regulated by the microenvironment. Nat Cell Biol 12(5):468–476. https://doi.org/10.1038/ncb2048

    Article  CAS  PubMed  Google Scholar 

  28. Werbowetski-Ogilvie TE, Bhatia M (2008) Pluripotent human stem cell lines: what we can learn about cancer initiation. Trends Mol Med 14(8):323–332. https://doi.org/10.1016/j.molmed.2008.06.005

    Article  CAS  PubMed  Google Scholar 

  29. Benoit YD, Witherspoon MS, Laursen KB, Guezguez A, Beausejour M, Beaulieu JF, Lipkin SM, Gudas LJ (2013) Pharmacological inhibition of polycomb repressive complex-2 activity induces apoptosis in human colon cancer stem cells. Exp Cell Res 319(10):1463–1470. https://doi.org/10.1016/j.yexcr.2013.04.006

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yannick D. Benoit .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Benoit, Y.D. (2018). Identification of Novel Molecules Targeting Cancer Stem Cells. In: Beaulieu, JF. (eds) Colorectal Cancer. Methods in Molecular Biology, vol 1765. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7765-9_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7765-9_22

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7764-2

  • Online ISBN: 978-1-4939-7765-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics