Limits...
Selective Targeting of CTNBB1-, KRAS- or MYC-Driven Cell Growth by Combinations of Existing Drugs.

Uitdehaag JC, de Roos JA, van Doornmalen AM, Prinsen MB, Spijkers-Hagelstein JA, de Vetter JR, de Man J, Buijsman RC, Zaman GJ - PLoS ONE (2015)

Bottom Line: We demonstrate that the Wnt-pathway inhibitor ICG-001 and trametinib acted synergistically in Wnt-pathway-mutant cell lines.The ERBB2 inhibitor TAK-165 was synergistic with trametinib in KRAS-mutant cell lines.The EGFR/ERBB2 inhibitor neratinib acted synergistically with the spindle poison docetaxel and with the Aurora kinase inhibitor GSK-1070916 in cell lines with MYC amplification.

View Article: PubMed Central - PubMed

Affiliation: Netherlands Translational Research Center B.V., Oss, The Netherlands.

ABSTRACT
The aim of combination drug treatment in cancer therapy is to improve response rate and to decrease the probability of the development of drug resistance. Preferably, drug combinations are synergistic rather than additive, and, ideally, drug combinations work synergistically only in cancer cells and not in non-malignant cells. We have developed a workflow to identify such targeted synergies, and applied this approach to selectively inhibit the proliferation of cell lines with mutations in genes that are difficult to modulate with small molecules. The approach is based on curve shift analysis, which we demonstrate is a more robust method of determining synergy than combination matrix screening with Bliss-scoring. We show that the MEK inhibitor trametinib is more synergistic in combination with the BRAF inhibitor dabrafenib than with vemurafenib, another BRAF inhibitor. In addition, we show that the combination of MEK and BRAF inhibitors is synergistic in BRAF-mutant melanoma cells, and additive or antagonistic in, respectively, BRAF-wild type melanoma cells and non-malignant fibroblasts. This combination exemplifies that synergistic action of drugs can depend on cancer genotype. Next, we used curve shift analysis to identify new drug combinations that specifically inhibit cancer cell proliferation driven by difficult-to-drug cancer genes. Combination studies were performed with compounds that as single agents showed preference for inhibition of cancer cells with mutations in either the CTNNB1 gene (coding for β-catenin), KRAS, or cancer cells expressing increased copy numbers of MYC. We demonstrate that the Wnt-pathway inhibitor ICG-001 and trametinib acted synergistically in Wnt-pathway-mutant cell lines. The ERBB2 inhibitor TAK-165 was synergistic with trametinib in KRAS-mutant cell lines. The EGFR/ERBB2 inhibitor neratinib acted synergistically with the spindle poison docetaxel and with the Aurora kinase inhibitor GSK-1070916 in cell lines with MYC amplification. Our approach can therefore efficiently discover novel drug combinations that selectively target cancer genes.

No MeSH data available.


Related in: MedlinePlus

Synergy of the MEK inhibitor trametinib (blue) and the Wnt-pathway inhibitor ICG-001 (green) in colon cancer cell lines.A: Synergy in CTNNB1-wild type cell line SW620. This cell line is nevertheless a Wnt-pathway-mutant as it is mutated in APC. In addition, SW620 is TP53-mutant. B: Absence of synergy in the CTNNB1-mutant cell line SW48. This cell line is TP53-wild type. C: isobologram of results in panel A, at the 75% effect cut-off level. Yellow, red, orange are mixture curves as described in Fig 1.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4446296&req=5

pone.0125021.g008: Synergy of the MEK inhibitor trametinib (blue) and the Wnt-pathway inhibitor ICG-001 (green) in colon cancer cell lines.A: Synergy in CTNNB1-wild type cell line SW620. This cell line is nevertheless a Wnt-pathway-mutant as it is mutated in APC. In addition, SW620 is TP53-mutant. B: Absence of synergy in the CTNNB1-mutant cell line SW48. This cell line is TP53-wild type. C: isobologram of results in panel A, at the 75% effect cut-off level. Yellow, red, orange are mixture curves as described in Fig 1.

Mentions: Synergistic interactions between trametinib and ICG-001 were investigated in a panel of cell lines, including three CTNNB1-wild type colon cancer cell lines, three CTNNB1-mutant colon cancer cell lines, and a pair of CTNNB1-mutant and CTNNB1-wild type lung cancer cell lines (Fig 8 and S3 Fig). Interestingly, and in contrast to our expectations, trametinib and ICG-001 appear to work synergistically in the CTNNB1-wild type colon cancer cell line SW620 (Fig 8A), and not in the CTNNB1-mutant cell line (Fig 8B). This is confirmed by isobologram analysis (Fig 8C). To generate more reliable CI averages, the experiments were repeated seven times, on several occasions by different scientists (Fig 8, S4 Table). the result is supported by experiments in other cell lines. In the CTNNB1-wild type colon cancer cell line HCT 15 synergy is observed, and not in other lines such as HCT 116 (mutant), LoVo (wild type) and LS174T (mutant, S3 Fig). In lung cancer cell lines, the result is more straightforward. Here the combination of ICG-001 and trametinib shows synergy in the CTNNB1-mutant cell line (A427, CI0.5 = 0.76, SD: 0.11) and not in the wild type cell line (A549, CI0.5 = 1.03, SD: 0.06, S3 Fig).


Selective Targeting of CTNBB1-, KRAS- or MYC-Driven Cell Growth by Combinations of Existing Drugs.

Uitdehaag JC, de Roos JA, van Doornmalen AM, Prinsen MB, Spijkers-Hagelstein JA, de Vetter JR, de Man J, Buijsman RC, Zaman GJ - PLoS ONE (2015)

Synergy of the MEK inhibitor trametinib (blue) and the Wnt-pathway inhibitor ICG-001 (green) in colon cancer cell lines.A: Synergy in CTNNB1-wild type cell line SW620. This cell line is nevertheless a Wnt-pathway-mutant as it is mutated in APC. In addition, SW620 is TP53-mutant. B: Absence of synergy in the CTNNB1-mutant cell line SW48. This cell line is TP53-wild type. C: isobologram of results in panel A, at the 75% effect cut-off level. Yellow, red, orange are mixture curves as described in Fig 1.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4446296&req=5

pone.0125021.g008: Synergy of the MEK inhibitor trametinib (blue) and the Wnt-pathway inhibitor ICG-001 (green) in colon cancer cell lines.A: Synergy in CTNNB1-wild type cell line SW620. This cell line is nevertheless a Wnt-pathway-mutant as it is mutated in APC. In addition, SW620 is TP53-mutant. B: Absence of synergy in the CTNNB1-mutant cell line SW48. This cell line is TP53-wild type. C: isobologram of results in panel A, at the 75% effect cut-off level. Yellow, red, orange are mixture curves as described in Fig 1.
Mentions: Synergistic interactions between trametinib and ICG-001 were investigated in a panel of cell lines, including three CTNNB1-wild type colon cancer cell lines, three CTNNB1-mutant colon cancer cell lines, and a pair of CTNNB1-mutant and CTNNB1-wild type lung cancer cell lines (Fig 8 and S3 Fig). Interestingly, and in contrast to our expectations, trametinib and ICG-001 appear to work synergistically in the CTNNB1-wild type colon cancer cell line SW620 (Fig 8A), and not in the CTNNB1-mutant cell line (Fig 8B). This is confirmed by isobologram analysis (Fig 8C). To generate more reliable CI averages, the experiments were repeated seven times, on several occasions by different scientists (Fig 8, S4 Table). the result is supported by experiments in other cell lines. In the CTNNB1-wild type colon cancer cell line HCT 15 synergy is observed, and not in other lines such as HCT 116 (mutant), LoVo (wild type) and LS174T (mutant, S3 Fig). In lung cancer cell lines, the result is more straightforward. Here the combination of ICG-001 and trametinib shows synergy in the CTNNB1-mutant cell line (A427, CI0.5 = 0.76, SD: 0.11) and not in the wild type cell line (A549, CI0.5 = 1.03, SD: 0.06, S3 Fig).

Bottom Line: We demonstrate that the Wnt-pathway inhibitor ICG-001 and trametinib acted synergistically in Wnt-pathway-mutant cell lines.The ERBB2 inhibitor TAK-165 was synergistic with trametinib in KRAS-mutant cell lines.The EGFR/ERBB2 inhibitor neratinib acted synergistically with the spindle poison docetaxel and with the Aurora kinase inhibitor GSK-1070916 in cell lines with MYC amplification.

View Article: PubMed Central - PubMed

Affiliation: Netherlands Translational Research Center B.V., Oss, The Netherlands.

ABSTRACT
The aim of combination drug treatment in cancer therapy is to improve response rate and to decrease the probability of the development of drug resistance. Preferably, drug combinations are synergistic rather than additive, and, ideally, drug combinations work synergistically only in cancer cells and not in non-malignant cells. We have developed a workflow to identify such targeted synergies, and applied this approach to selectively inhibit the proliferation of cell lines with mutations in genes that are difficult to modulate with small molecules. The approach is based on curve shift analysis, which we demonstrate is a more robust method of determining synergy than combination matrix screening with Bliss-scoring. We show that the MEK inhibitor trametinib is more synergistic in combination with the BRAF inhibitor dabrafenib than with vemurafenib, another BRAF inhibitor. In addition, we show that the combination of MEK and BRAF inhibitors is synergistic in BRAF-mutant melanoma cells, and additive or antagonistic in, respectively, BRAF-wild type melanoma cells and non-malignant fibroblasts. This combination exemplifies that synergistic action of drugs can depend on cancer genotype. Next, we used curve shift analysis to identify new drug combinations that specifically inhibit cancer cell proliferation driven by difficult-to-drug cancer genes. Combination studies were performed with compounds that as single agents showed preference for inhibition of cancer cells with mutations in either the CTNNB1 gene (coding for β-catenin), KRAS, or cancer cells expressing increased copy numbers of MYC. We demonstrate that the Wnt-pathway inhibitor ICG-001 and trametinib acted synergistically in Wnt-pathway-mutant cell lines. The ERBB2 inhibitor TAK-165 was synergistic with trametinib in KRAS-mutant cell lines. The EGFR/ERBB2 inhibitor neratinib acted synergistically with the spindle poison docetaxel and with the Aurora kinase inhibitor GSK-1070916 in cell lines with MYC amplification. Our approach can therefore efficiently discover novel drug combinations that selectively target cancer genes.

No MeSH data available.


Related in: MedlinePlus