Resistant Cancer Cell Line Collection

Jindrich Cinatl
Dr Petra Joh-Forschungshaus
Frankfurter Stiftung für krebskranke Kinder
Frankfurt am Main, Germany

Wass-Michaelis Research Group
School of Biosciences
University of Kent
Kent, UK
About the Resistant Cancer Cell Line Collection Database
A novel resource for the studying of acquired drug resistance in cancer and the development of novel anti-cancer therapies
Background - Acquired drug resistance in Cancer

The formation of acquired drug resistance is a major reason for the failure of anti-cancer therapies. Many cancers initially respond well to drug treatment but eventually resistant cancer cells emerge resulting in therapy failure and patient death. It is important to understand this process to enable the improved methods for detecting resistance occuring and to identify suitable next line therapies.

it has been shown that the resistance mechanisms underlying acquired resistance can substantially differ from those underlying intrinsic resistance [see e.g. Hata et al., 2016; Carter et al., 2017], pre-clinical model systems of acquired drug resistance are needed to fill this gap. Pre-clinical models enable 1) the detailed systems level monitoring of processes in cancer cell populations in response to therapy over time (that is limited in the clinical setting due to intra-tumour heterogeneity [Zardavas et al., 2015; Esposito et al., 2016]), 2) the study of a large number of different cancer treatments (that is clinically limited by patient numbers and patients’ rights to the best available therapy), and 3) extensive functional studies including the  comparative testing of different therapy regimens in a given cancer population (clinically, every individual patient can only be treated once).

To systematically study acquired drug resistance in cancer, we have now established the Resistant Cancer Cell Line (RCCL) collection, a unique collection of 1300 cancer cell lines representing acquired resistance in 15 cancer entities and includes cell lines adapted to 67 targeted and cytotoxic anti-cancer drugs. The full lost of cell lines in the RCCL Collection is available here.

 
Using drug adapted cell lines as a model for acquired drug resistance in cancer

Cancer cell lines are a leading preclinical cancer model system [Sharma et al., 2010]. They are easy to handle and manipulate and enable the generation of the large number of individual models needed to cover the complexity of the resistance formation process. Major resistance mechanisms have been discovered in drug-adapted cell lines, e.g. the ATP-binding cassette (ABC) transporters ABCB1 (also known as P-glycoprotein or MDR1) [Juliano & Ling, 1976] and ABCC1 (also known as MRP1) [Cole et al., 1992]. Moreover, drug-adapted cancer cell lines have been successfully used by different research groups to identify and investigate clinically relevant acquired resistance mechanisms to targeted [Engelman et al., 2007; Nazarian et al., 2010; Poulikakos et al., 2011; Joseph et al., 2013; Korpal et al., 2013; Crystal et al., 2014; Hata et al., 2016] and cytotoxic [Domingo-Domenech et al., 2012; Zahreddine et al., 2014; Göllner et al., 2017] anti-cancer drugs.

Using drug-adapted cancer cell lines derived from the RCCL collection, we have already shown that acquisition of resistance to MDM2 inhibitors is due to the formation of de novo p53 mutations [Michaelis et al., 2011; 2012], and this has recently been confirmed clinically [Jung et al., 2016]. In addition, a screen in a panel of drug-adapted urothelial cancer cell lines identified the vinca alkaloids vinblastine and vinflunine, the approved second-line therapeutic for metastatic urothelial cancer, as the most effective compounds [Vallo et al., 2015]. Most recently, we have also demonstrated that high SAMHD1 expression is a resistance mechanism in cytarabine-adapted  acute myeloid leukaemia cell lines. Based on this discovery, we have shown that SAMHD1 is a biomarker for cytarabine response in the clinic  and a novel therapeutic target in acute myeloid leukaemia [Schneider et al., 2017].

Establishing drug adapted cancer cell lines in the RCCL Collection
The drug adaptation process

In the drug adaptation process, a drug sensitive cell line is exposed to a drug, typically using a process of dose escalation i.e. initially growing the cells in a low dose of drug and gradually increasing the concentration over time (as shown in the figure above). For some cell lines this can take more than a year.

Once the cell have become resistant to the drug (typically with an IC50 more than two fold greater than the parental cell line), they can be used a model of acquired drug resistance in cancer. The study of these cell lines can then be used to investigate 1) mechanisms of drug resistance, sensitivity to other drugs and biomarker of acquired resistance.

The current release of the RCCL Collection Database
Some statistics about the current release
Cell line drug combinations1980
Cancer types11
Total cell lines339
parental cell lines49
drug adapted cell lines290
drugs for which sensitivity is tested100
drug classes9
Number of studies29
Number of published studies23
Number of unpublished studies6

Data in the current release has been published in the following papers

Treatment of drug-resistant human neuroblastoma cells with cyclodextrin inclusion complexes of aphidicolin
Michaelis M, Cinatl J, Vogel JU, Pouckova P, Driever PH, Cinatl J Jr., Anticancer Drugs. 2001 12:467-73.
Increased malignant behavior in neuroblastoma cells with acquired multi-drug resistance does not depend on P-gp expression
Kotchetkov R, Driever PH, Cinatl J, Michaelis M, Karaskova J, Blaheta R, Squire JA, Von Deimling A, Moog J, Cinatl J Jr., Int J Oncol. 2005 27:1029-37.
Anti-cancer effects of bortezomib against chemoresistant neuroblastoma cell lines in vitro and in vivo.
Michaelis M, Fichtner I, Behrens D, Haider W, Rothweiler F, Mack A, Cinatl J, Doerr HW, Cinatl J Jr., Int J Oncol. 2006 28:439-46.
Onconase induces caspase-independent cell death in chemoresistant neuroblastoma cells
Michaelis M, Cinatl J, Anand P, Rothweiler F, Kotchetkov R, von Deimling A, Doerr HW, Shogen K, Cinatl J Jr., Cancer Lett. 2007 250:107-16.
Cisplatin-resistant neuroblastoma cells express enhanced levels of epidermal growth factor receptor (EGFR) and are sensitive to treatment with EGFR-specific toxins
Michaelis M, Bliss J, Arnold SC, Hinsch N, Rothweiler F, Deubzer HE, Witt O, Langer K, Doerr HW, Wels WS, Cinatl J Jr., Clin Cancer Res. 2008 14:6531-7.
Reversal of P-glycoprotein-mediated multidrug resistance by the murine double minute 2 antagonist nutlin-3
Michaelis M, Rothweiler F, Klassert D, von Deimling A, Weber K, Fehse B, Kammerer B, Doerr HW, Cinatl J Jr., Cancer Res. 2009 69:416-21.
Anti-cancer effects of artesunate in a panel of chemoresistant neuroblastoma cell lines.
Michaelis M, Kleinschmidt MC, Barth S, Rothweiler F, Geiler J, Breitling R, Mayer B, Deubzer H, Witt O, Kreuter J, Doerr HW, Cinatl J, Cinatl J Jr., Biochem Pharmacol. 2010 79:130-6.
Anticancer effects of the nitric oxide-modified saquinavir derivative saquinavir-NO against multidrug-resistant cancer cells.
 Rothweiler F, Michaelis M, Brauer P, Otte J, Weber K, Fehse B, Doerr HW, Wiese M, Kreuter J, Al-Abed Y, Nicoletti F, Cinatl J Jr.,Neoplasia. 2010 12:1023-30.
Adaptation of cancer cells from different entities to the MDM2 inhibitor nutlin-3 results in the emergence of p53-mutated multi-drug-resistant cancer cells.
Michaelis M, Rothweiler F, Barth S, Cinatl J, van Rikxoort M, Löschmann N, Voges Y, Breitling R, von Deimling A, Rödel F, Weber K, Fehse B, Mack E, Stiewe T, Doerr HW, Speidel D,Cinatl J Jr., Cell Death Dis. 2011 2:e243.
Human neuroblastoma cells with acquired resistance to the p53 activator RITA retain functional p53 and sensitivity to other p53 activating agents
Michaelis M, Rothweiler F, Agha B, Barth S, Voges Y, Löschmann N, von Deimling A, Breitling R, Doerr HW, Rödel F, Speidel D, Cinatl J Jr., Cell Death Dis. 2012 3:e294.
Testing of SNS-032 in a Panel of Human Neuroblastoma Cell Lines with Acquired Resistance to a Broad Range of Drugs
 Löschmann N, Michaelis M, Rothweiler F, Zehner R, Cinatl J, Voges Y, Sharifi M, Riecken K, Meyer J, von Deimling A, Fichtner I, Ghafourian T, Westermann F,Cinatl J Jr., Transl Oncol. 2013 6:685-96.
Aurora kinases as targets in drug-resistant neuroblastoma cells
Michaelis M, Selt F, Rothweiler F, Löschmann N, Nüsse B, Dirks WG, Zehner R, Cinatl J Jr., PLoS One. 2014 9:e108758.
Association between acquired resistance to PLX4032 (vemurafenib) and ATP-binding cassette transporter expression
Michaelis M, Rothweiler F, Nerreter T, van Rikxoort M, Zehner R, Dirks WG, Wiese M,Cinatl J Jr., BMC Res Notes. 2014 7:710.
Identification of flubendazole as potential anti-neuroblastoma compound in a large cell line screen
Michaelis M, Michaelis M, Agha B, Rothweiler F, Löschmann N, Voges Y, Mittelbronn M, Starzetz T, Harter PN, Abhari BA, Fulda S, Westermann F, Riecken K, Spek S, Langer K, Wiese M, Dirks WG, Zehner R, Cinatl J, Wass MNCinatl J Jr., Sci Rep. 2015 5:8202.
Enzastaurin inhibits ABCB1-mediated drug efflux independently of effects on protein kinase C signalling and the cellular p53 status.
Michaelis M, Rothweiler F, Löschmann N, Sharifi M, Ghafourian T,Cinatl J Jr., Oncotarget. 2015 6:17605-20.
Drug-Resistant Urothelial Cancer Cell Lines Display Diverse Sensitivity Profiles to Potential Second-Line Therapeutics.
Vallo S, Michaelis M, Rothweiler F, Bartsch G, Gust KM, Limbart DM, Rödel F, Wezel F, Haferkamp A Cinatl J Jr., Transl Oncol. 2015 8:210-6.
Substrate-specific effects of pirinixic acid derivatives on ABCB1-mediated drug transport
Michaelis M, Rothweiler F, Wurglics M, Aniceto N, Dittrich M, Zettl H, Wiese M, Wass M, Ghafourian T, Schubert-Zsilavecz M, Cinatl J Jr., Oncotarget. 2016 7:11664-76.
ABCB1 as predominant resistance mechanism in cells with acquired SNS-032 resistance
 Löschmann N, Michaelis M, Rothweiler F, Voges Y, Balónová B, Blight BA, Cinatl J Jr., Oncotarget. 2016 7:58051-58064.
Dasatinib enhances tumor growth in gemcitabine-resistant orthotopic bladder cancer xenografts
 Vallo S, Michaelis M, Gust KM, Black PC, Rothweiler F, Kvasnicka HM, Blaheta RA, Brandt MP, Wezel F, Haferkamp A,Cinatl J Jr., BMC Res Notes. 2016 9:454.
Effects of YM155 on survivin levels and viability in neuroblastoma cells with acquired drug resistance
 Voges Y, Michaelis M, Rothweiler F, Schaller T, Schneider C, Politt K, Mernberger M, Nist A, Stiewe T, Wass MN, Rödel F, Cinatl J.Cinatl J Jr., Anticancer Drugs. 2001 12:467-73.
SAMHD1 is a biomarker for cytarabine response and a therapeutic target in acute myeloid leukemia
Schneider C, Oellerich T, Baldauf HM, Schwarz SM, Thomas D, Flick R, Bohnenberger H, Kaderali L, Stegmann L, Cremer A, Martin M, Lohmeyer J, Michaelis M, Hornung V, Schliemann C, Berdel WE, Hartmann W, Wardelmann E, Comoglio F, Hansmann ML, Yakunin AF, Geisslinger G, Ströbel P, Ferreirós N, Serve H, Keppler OT, Cinatl J Jr., Nat Med. 2017 23:250-255.
Acquired resistance to oxaliplatin is not directly associated with increased resistance to DNA damage in SK-N-ASrOXALI4000, a newly established oxaliplatin-resistant sub-line of the neuroblastoma cell line SK-N-AS
Saintas E, Abrahams L, Ahmad GT, Ajakaiye AM, AlHumaidi AS, Ashmore-Harris C, Clark I, Dura UK, Fixmer CN, Ike-Morris C, Mato Prado M, Mccullough D, Mishra S, Schöler KM, Timur H, Williamson MD, Alatsatianos M, Bahsoun B, Blackburn E, Hogwood CE, Lithgow PE, Rowe M, Yiangou L, Rothweiler F, Cinatl J Jr, Zehner R, Baines AJ, Garrett MD, Gourlay CW, Griffin DK, Gullick WJ, Hargreaves E, Howard MJ, Lloyd DR, Rossman JS, Smales CM, Tsaousis AD, von der Haar T, Wass MNMichaelis M. PLoS One. 2017 12:e0172140.
Resistance to nanoparticle albumin-bound paclitaxel is mediated by ABCB1 in urothelial cancer cells
Vallo S, Köpp R, Michaelis M, Rothweiler F, Bartsch G, Brandt MP, Gust KM, Wezel F, Blaheta RA, Haferkamp A,Cinatl J Jr., Oncol Lett. 2017 13:4085-4092.