NTRC expands Oncolines™ cell line panel to 102 cell lines

Oss, June 22nd 2017 – Today NTRC has launched an expansion of its Oncolines™ cancer cell line panel, from 66 to 102 cell lines. Oncolines™ are proliferation assays on a panel of genetically well-characterized cancer cell lines from diverse tumor tissue origin. NTRC has a commercial license from the American Type Culture Collection (ATCC). Oncolines™ profiling includes genomic biomarker analysis and comparative profiling against a library of more than 150 pre-profiled anti-cancer agents. In recent publications in Molecular Cancer Therapeutics [1] and Nature Communications [2] the reproducibility of Oncolines™ cell panel profiling and its application in mechanism-of-action studies has been demonstrated. The Oncolines™ biomarker analysis can be extended with gene expression analysis through GeneNominator™.

Characteristics Oncolines™:
– Readout ATPlite™
– Low passage number and ATCC recommended culturing conditions
– No IC50 extrapolation
– 384-well, automated liquid handling
– Manual inspection of curves

Recent publications with Oncolines™ and OncolinesProfiler™ results:
[1] Uitdehaag et al. (2016) Cell panel profiling reveals conserved therapeutic clusters and differentiates the mechanism of action of different PI3K/mTOR, Aurora kinase and EZH2 inhibitors. Molecular Cancer Therapeutics 15, 3097-3109
[2] Bohnacker et al. (2017) Deconvolution of Buparlisib’s mechanism of action defines specific PI3K and tubulin inhibitors for therapeutic intervention. Nature Communication 8:14683

For more information or quotations, please sent e-mail to services@ntrc.nl

About NTRC
NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

NTRC publishes target residence time-guided optimization of TTK inhibitors

Oss, May 22nd 2017 – This week, NTRC published results from its TTK (Mps1) drug discovery programme in the Journal of Molecular Biology. TTK is a promising new drug target for the treatment of aggressive cancers such as triple-negative breast cancer. NTRC combined the ResidenceTimer™ platform for determination of binding kinetics with protein crystallography to develop inhibitors with a unique mechanism of action. Compared to reference inhibitors, NTRC’s TTK inhibitors bind in a unique ‘lysine trap’ binding mode that shifts the so-called glycine rich loop in TTK and locks the catalytic lysine in a dysfunctional state. As a result of these stabilizing rearrangements, the inhibitors have long target residence times which translate into very potent antiproliferative activities that rival those of classic cytotoxic therapy. This was measured in NTRC’s Oncolines™ cancer cell line panel. The unique properties and potent activity open up new and specific applications for TTK inhibitors in cancer treatment. To access the article follow: http://dx.doi.org/10.1016/j.jmb.2017.05.014

About NTRC

NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, GeneNominator™ and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

NTRC launches GeneNominator™ tool for drug sensitivity and gene expression analysis

Oss, March 30th, 2017 – NTRC announces its GeneNominator™ bioinformatics package to study the biology of inhibitor response at the mRNA expression level. With NTRC’s Oncolines™ panel, the IC50 values of inhibitors can be determined in a rapid and reproducible fashion. With the new GeneNominator™ tool, these IC50 values can be connected to the expression levels of more than 18,000 individual genes in each of the cell lines. This enables identification of pathways that confer sensitivity or resistance to customer inhibitors. False positive scoring genes are filtered out by comparing responses with a proprietary database of response profiles of 150 different drugs and drug candidates. Interesting genes are highlighted through the use of relevant subsets. GeneNominator™ makes use of state-of-the-art analyses such as protein-protein-interaction predictions and Gene Set Analysis to give meaning to gene lists and identify the important pathways in responsive and resistant cell populations. This includes the prediction of responder populations at the clinical level.
GeneNominator™ will be presented at the NTRC booth (#3050) at the American Association of Cancer Research 2017 Annual meeting in Washington D.C. For more information: info@ntrc.nl.

GeneNominator_PR.png
MDM2 gene expression in cell lines, which inhibits TP53 activity, strongly correlates with sensitivity for the MDM2 inhibitor nutlin-3a (left). This is complementary to the Oncolines™ genomic biomarker association which shows that TP53 wild type cell lines are sensitive for nutlin-3a (right).

About NTRC
NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

Meet NTRC at booth #3050 AACR 2017 in Washington D.C.

Oss, March, 27th, 2017 – NTRC will exhibit its services at the AACR conference 2017, to be held in Washington D.C. next week. Please be welcome at booth #3050 in the Exhibition Hall.

 

Two posters on the TTK kinase drug discovery program will be presented on Tuesday, April 4th.

Poster Titles and Sessions:

Title: Stable aneuploid cells are more sensitive to TTK inhibition than chromosome instable cell lines

Session Title: Mitosis, Telomeres, and Proliferation Control Session

Date & time: Tuesday April 4th, 8:00 am – 12:00pm

Location: Poster Section #19, Poster Board #7, Poster #3457

Presenter: Marion Libouban

 

Title: NTRC 1501-0, a TTK kinase inhibitor selected for its long target residence time, completely inhibits tumor growth in the MDA-MB-231 xenograft model for triple-negative breast cancer

Session Title: Targeting Protein Kinases and DNA Repair

Date & time: Tuesday April 4th, 1:00 – 5:00pm

Location: Poster Section #7, Poster Board #8, Poster #4185

Presenter: Joost Uitdehaag

 

About NTRC

NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

 

New study in Oncotarget shows that stable aneuploid cells are more sensitive to TTK inhibition than chromosomally unstable cell lines

Oss, March, 16th, 2017 – In a research article which appeared online today in Oncotarget, NTRC scientists, in collaboration with Prof. Dr. René Medema from the Netherlands Cancer Institute (NKI), show that stable aneuploid tumor cells are more sensitive to small molecule inhibitors of TTK than chromosome instable cells. Chromosomal instability is a hallmark of cancer and one of the main causes of “aneuploidy”, a state of an imbalanced chromosome number. The protein kinase TTK, commonly referred to as Mps1, is a component of the spindle assembly checkpoint, a surveillance mechanism that controls the fidelity of chromosome segregation. High levels of TTK correlate with chromosomal instability in several aggressive cancers, such as breast cancer and hepatocellular carcinoma. To determine the relationship between chromosomal instability and sensitivity to TTK inhibitors, Dr. Marion Libouban from NTRC investigated the effect of TTK inhibition on cancer cells with abnormal chromosomal states using karyotype analysis and time-lapse microscopy at NKI. Time-lapse imaging is a live cell imaging technique that allows visualization of the mitotic timing and chromosomal mis-segregation during the metaphase to anaphase transition.

Taking advantage of the selective and sub-nanomolar potent TTK inhibitor NTRC 0066-0, which was developed by NTRC {link to article in Annals of Oncology}, Libouban showed that inhibition of TTK overrides the mitotic checkpoint, irrespective of cell line sensitivity. In stable aneuploid cells, such as the colorectal adenocarcinoma cell line LoVo (see photograph), NTRC 0066-0 induced acute chromosomal instability. In contrast, in cells with high levels of pre-existing chromosomal instability there was only a small additional fraction of cells mis-segregating their chromosomes. In proliferation assays stable aneuploid cells were more sensitive than cell lines with pre-existing chromosomal instability.

Lovo.png

NTRC 0066-0 induces massive chromosome mis-segregation in LoVo cells. The left panel shows vehicle-treated cells as control.

In collaboration with Prof. Dr. Zuzana Storchova from the Technical University Kaiserslautern (Germany), Libouban and colleagues investigated the effect of TTK inhibitors on post-tetraploid cancer cells. Tetraploids are thought to be an intermediate between diploid and unstable aneuploid cells. More than a third of human cancers went through tetraploidy during tumorigenesis. NTRC scientists, in collaboration with Prof. Storchova, have previously shown that post-tetraploids are more resistant to multiple cytotoxic drugs and targeted agents {link to article in Cell Cycle}. Remarkably, as shown in the Oncotarget article, TTK inhibitors had the same potency on post-tetraploid and parental diploid cells, suggesting that TTK targeting may be a better choice for the eradication of tumor cells that underwent whole genome doubling than other drugs interfering with mitosis.

For the first time, the effect of a TTK inhibitor on the viability and proliferation of primary human patient-derived tumor cell samples and organoids was studied. NTRC 0066-0 inhibited the proliferation of patient-derived colorectal cancer organoids with a potency similar to that of cancer cell lines. In contrast, NTRC 0066-0 did not reduce the viability of non-proliferating T cell acute lymphoblastic leukemia cell samples. Consequently, TTK inhibitor therapy is expected to spare non-dividing cells, and may be used to target chromosomally stable aneuploid tumors.

To access the article follow: link to Oncotarget

The study was performed in the frame-work of the European Commission-funded FP7 Marie Curie International Training Network project “PloidyNet

About NTRC

NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

 

Kinetic parameters of 80 kinase inhibitors available online through the Journal of Molecular Biology

Oss, January, 2nd, 2017 – NTRC has made available the kinetic data of 80 kinase inhibitors on their targets in a research article in the Journal of Molecular Biology. This is the largest data base of association and dissociation rate constants of kinase inhibitors available to date. In total, 46 protein tyrosine, serine/threonine and lipid kinases were studied, including well-known drug targets such as the epidermal growth factor receptor (EGFR) and the δ-isoform of phosphoinositide 3-kinase (PI3Kδ). Kinetic constants were determined on NTRC’s ResidenceTimer™ platform using recombinant kinases from Carna Biosciences, Inc. (Kobe, Japan).

Kinases are the target of more than thirty registered small molecule drugs, while more than 300 kinase inhibitors are currently investigated in clinical studies. With the growing interest in the targeting of kinases in non-cancer indications and chronic diseases, there is an increased demand for selective inhibitors. The relative selectivity of kinase inhibitors is commonly determined by measuring their inhibitory potency (IC50) in a large panel of enzyme assays. These assays are usually performed at different ATP concentrations and with different substrates, in a closed system with a fixed concentration of inhibitor. In living systems the concentration of inhibitors is continuously changing. The time a compound resides on its target, or ‘target residence time’ (τ), is therefore a better predictor of its biological activity than its IC50 in enzyme assays. Thus, compared to traditional enzyme assays, ResidenceTimer™ measurements allow the determination of the binding kinetics and selectivity of kinase inhibitors in a system that better resembles real life biology than assays in equilibrium or end-point assays.

In the article in Journal of Molecular Biology a number of comparative studies are presented for, a.o., different Aurora and PI3K inhibitors. These case studies provide clear arguments to include target residence time measurements in kinase inhibitor drug discovery programs. To access the article follow: http://dx.doi.org/10.1016/j.jmb.2016.12.019

About NTRC

NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

Structural insight into the function of TDO in healthy aging

Oss, Groningen, December, 23rd, 2016 – Researchers from the European Research Institute for the Biology of Aging (ERIBA) University of Groningen, the Netherlands) in collaboration with the Netherlands Translational Research Center B.V. have identified an evolutionary conserved structural loop in the enzyme TDO that is critical for its function. When absent, it increases the motility and the lifespan of the roundworm Caenorhabditis elegans. The structural element was identified in a genetic screen by the group of Dr. Ellen Nollen at ERIBA. In a previous study, Nollen and colleagues showed that in C. elegans, depletion of TDO by RNA interference suppresses the toxicity from aggregation of the protein α-synuclein and extended lifespan of the worms. In humans α-synuclein aggregation has been implicated in Parkinson’s disease. In the new study they now report the remarkable finding that deletion of only three amino acids in TDO has the same effect as deletion of the whole gene. The study appeared in the online open access journal Scientific Reports, and includes 3D homology modelling studies performed at NTRC, which suggest that the three amino acid motif is involved in the binding of haem, an essential co-factor of TDO. Furthermore, the motif is conserved throughout evolution. In humans the motif is likely to be involved in the interaction with small molecule inhibitors. Such molecules are currently being developed by several companies, including NTRC, and have potential therapeutic application in Parkinson’s disease and certain cancers that express TDO.

To access the article, please follow: www.nature.com/articles/srep39199

About NTRC

NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

New drug combinations addressing treatment resistance in childhood leukemia proposed based on SynergyFinder™

Utrecht, Oss, December, 20th, 2016 – Researchers from the Princess Máxima Center for Pediatric Oncology in Utrecht (The Netherlands) in collaboration with researchers from Erasmus Medical Center in Rotterdam have identified mechanisms underlying resistance to steroid therapy in childhood T-cell acute lymphoblastic leukemia (T-ALL). In the article, which appeared today in the open access online journal PLOS Medicine, new approaches to therapy of childhood T-ALL are proposed based on SynergyFinder™ studies performed at NTRC.

Treatment of childhood leukemia has improved markedly in recent decades, with long-term cure achieved in most patients who have access to modern treatment regimens. However, some patients develop resistance to the steroid drugs which are a key part of combination chemotherapy treatments, which results in poor clinical outcomes. Dr. Jules Meijerink, Principal Investigator at the recently established Princess Máxima Center, studied together with colleagues the causes of resistance by whole genome sequencing on paired samples collected at diagnosis and at remission of thirteen patients. The study was expanded by targeted exome sequencing of candidate genes in samples from 146 additional T-ALL patients. Specific gene mutations in the interleukin 7 (IL7) receptor and its downstream signalling molecules JAK1 and K-RAS were found to associate with steroid resistance and poor clinical outcome. The mutations did not change the functioning of the steroid receptor, but resulted in a strong activation of MEK-ERK and AKT, which are downstream signalling components in the IL7 receptor pathway. To address clinical relevance, primary T-ALL cell samples from eleven patients were analysed in SynergyFinder™ at NTRC. Inhibitors of IL7 receptor signalling, including inhibitors of MEK, AKT and mTOR, synergistically increased steroid-induced leukemic cell death. Discussing the research in an accompanying Perspective article in PLOS Medicine, Drs. Steven Goossens and Peter van Vlierberghe from the Cancer Research Center Ghent (Belgium) conclude that “inhibition of MEK-ERK or PI3K/AKT/mTOR signalling could enhance steroid sensitivity in T-ALL and potentially improve patient treatment outcome, a notion that warrants investigation in future prospective clinical trials.”

To access the article, please follow: http://journals.plos.org/plosmedicine/article?id=10.1371/journal.pmed.1002200

About NTRC

NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

 

NTRC’s Oncolines™ profiling is cover story in December issue of Molecular Cancer Therapeutics

Oss, December, 5th, 2016 – An analysis by NTRC of 122 anti-cancer agents is cover story of Molecular Cancer Therapeutics, a leading journal on cancer drug discovery from the American Association of Cancer Research (AACR). The article on comparative profiling is marked by the editors as one of the highlights of the December issue, which appeared in print last Friday. In the article NTRC scientists  describe the comparative analysis of the anti-proliferative activity of 122 publicly known anti-cancer agents in the Oncolines™ cell panel. The compounds studied include many recently approved targeted therapies, cytotoxic agents, and epigenetic modulators. For 38 compounds this is the first cancer cell panel profiling study. Good correlations were observed with other cell panels. This is illustrated with the cover image, which shows a Pearson correlation matrix of 51 anti-cancer agents that have been analysed in Oncolines™ panel and in the cell panel from the National Cancer Institute (NCI-60).

In the article on page 3097 Uitdehaag and colleagues show that the 122 anti-cancer agents can be arranged in twenty-six clusters of compounds with common mechanisms of action. This ‘hierarchical clustering’ includes nine clusters that have not been reported before, such as clusters of inhibitors of EZH2, bromodomain (BET) and the spindle assembly checkpoint kinase TTK (Mps1). The lack of clustering of several Aurora and PI3K inhibitors could be explained by differences in biochemical selectivity. Different clustering of PI3K and mTOR inhibitors corresponded with different genomic targeting. The study shows that cell line profiling is an excellent tool for the unbiased classification of anti-proliferative compounds.

Comparative cell panel profiling (OncolinesProfiler™) is part of the Oncolines™ cancer cell line profiling service of NTRC. Comparative profiling can provide more insight into the mechanism of action of new candidate drugs, and can be used to determine similarity or differentiation of new compounds from existing anti-cancer agents.

Reference: Uitdehaag et al. (2016) Cell panel profiling reveals conserved therapeutic clusters and differentiates the mechanism of action of different PI3K/mTOR, Aurora kinase and EZH2 inhibitors. Molecular Cancer Therapeutics 15 (12), 3097-3109.

To access the article, please follow:
http://mct.aacrjournals.org/cgi/content/abstract/1535-7163.MCT-16-0403

About NTRC

NTRC is a precision medicine company dedicated to the development of new anti-cancer drugs. NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, OncolinesProfiler™, and SynergyFinder™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl

NTRC presents on ResidenceTimer™ and OncolinesProfiler™ at 28th EORTC-NCI-AACR symposium in Munich

The 28th EORTC-NCI-AACR symposium on Molecular targets and Cancer therapeutics will take place from November 29th to December 2nd, 2016 in Munich, Germany. NTRC will present two posters: one on the application target residence time measurements for protein kinases (ResidenceTimer™), with case studies on PI3 Kinases and Aurora kinases; and one on comparative cancer cell line profiling; the other on comparative cancer cell line profiling (OncolinesProfiler™), with case studies on epigenetic modulators, classic chemotherapeutic agents and protein kinase inhibitors.

Poster titles and sessions:

‘Cell panel profiling of pre-clinical and clinical anti-cancer agents reveals conserved therapeutic clusters and differentiates the mechanism of action of different PI3K/mTOR, Aurora kinase and EZH2 inhibitors’

Presented by Dr. Joost Uitdehaag

Date: Tuesday 29 November

Poster Session: Molecular targeted agents I

Abstract number 52; Poster board number: 023

 

‘Compound selectivity and target residence time of kinase inhibitors studied with surface plasmon resonance’

Presented by Ms. Nicole Willemsen-Seegers

Date: Wednesday 30 November

Poster Session: Drug Screening

Abstract number: 235; Poster board number: 061.

 

Information on the technology platform of NTRC can be found at www.ntrc.nl/services.

 

NTRC facilitates the development of novel therapies by providing cancer cell line profiling services (Oncolines™, SynergyFinder™ and OncolinesProfiler™) and target residence time measurements for protein kinases (ResidenceTimer™) on a fee-for-service basis. NFK GreenScreen™, an assay read-out for the cancer immunotherapy drug targets IDO1 and TDO, is supplied to clients globally. For more information please visit www.ntrc.nl or contact info@ntrc.nl.