Systematic screening identifies ABCG2 as critical factor underlying synergy of kinase inhibitors with transcriptional CDK inhibitors
Abstract Background Triple-negative breast cancer (TNBC) is a subtype of breast cancer with limited treatment options and poor clinical prognosis. Inhibitors of transcriptional CDKs are currently under thorough investigation for application in the treatment of multiple cancer types, including breast...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-05-01
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| Series: | Breast Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13058-023-01648-x |
| _version_ | 1797831806099128320 |
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| author | Vera E. van der Noord Wanda van der Stel Gijs Louwerens Danielle Verhoeven Hendrik J. Kuiken Cor Lieftink Melanie Grandits Gerhard F. Ecker Roderick L. Beijersbergen Peter Bouwman Sylvia E. Le Dévédec Bob van de Water |
| author_facet | Vera E. van der Noord Wanda van der Stel Gijs Louwerens Danielle Verhoeven Hendrik J. Kuiken Cor Lieftink Melanie Grandits Gerhard F. Ecker Roderick L. Beijersbergen Peter Bouwman Sylvia E. Le Dévédec Bob van de Water |
| author_sort | Vera E. van der Noord |
| collection | DOAJ |
| description | Abstract Background Triple-negative breast cancer (TNBC) is a subtype of breast cancer with limited treatment options and poor clinical prognosis. Inhibitors of transcriptional CDKs are currently under thorough investigation for application in the treatment of multiple cancer types, including breast cancer. These studies have raised interest in combining these inhibitors, including CDK12/13 inhibitor THZ531, with a variety of other anti-cancer agents. However, the full scope of these potential synergistic interactions of transcriptional CDK inhibitors with kinase inhibitors has not been systematically investigated. Moreover, the mechanisms behind these previously described synergistic interactions remain largely elusive. Methods Kinase inhibitor combination screenings were performed to identify kinase inhibitors that synergize with CDK7 inhibitor THZ1 and CDK12/13 inhibitor THZ531 in TNBC cell lines. CRISPR-Cas9 knockout screening and transcriptomic evaluation of resistant versus sensitive cell lines were performed to identify genes critical for THZ531 resistance. RNA sequencing analysis after treatment with individual and combined synergistic treatments was performed to gain further insights into the mechanism of this synergy. Kinase inhibitor screening in combination with visualization of ABCG2-substrate pheophorbide A was used to identify kinase inhibitors that inhibit ABCG2. Multiple transcriptional CDK inhibitors were evaluated to extend the significance of the found mechanism to other transcriptional CDK inhibitors. Results We show that a very high number of tyrosine kinase inhibitors synergize with the CDK12/13 inhibitor THZ531. Yet, we identified the multidrug transporter ABCG2 as key determinant of THZ531 resistance in TNBC cells. Mechanistically, we demonstrate that most synergistic kinase inhibitors block ABCG2 function, thereby sensitizing cells to transcriptional CDK inhibitors, including THZ531. Accordingly, these kinase inhibitors potentiate the effects of THZ531, disrupting gene expression and increasing intronic polyadenylation. Conclusion Overall, this study demonstrates the critical role of ABCG2 in limiting the efficacy of transcriptional CDK inhibitors and identifies multiple kinase inhibitors that disrupt ABCG2 transporter function and thereby synergize with these CDK inhibitors. These findings therefore further facilitate the development of new (combination) therapies targeting transcriptional CDKs and highlight the importance of evaluating the role of ABC transporters in synergistic drug–drug interactions in general. |
| first_indexed | 2024-04-09T13:58:41Z |
| format | Article |
| id | doaj.art-3cd3539eec0948958a3e708b71d4da89 |
| institution | Directory Open Access Journal |
| issn | 1465-542X |
| language | English |
| last_indexed | 2024-04-09T13:58:41Z |
| publishDate | 2023-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Breast Cancer Research |
| spelling | doaj.art-3cd3539eec0948958a3e708b71d4da892023-05-07T11:27:39ZengBMCBreast Cancer Research1465-542X2023-05-0125112010.1186/s13058-023-01648-xSystematic screening identifies ABCG2 as critical factor underlying synergy of kinase inhibitors with transcriptional CDK inhibitorsVera E. van der Noord0Wanda van der Stel1Gijs Louwerens2Danielle Verhoeven3Hendrik J. Kuiken4Cor Lieftink5Melanie Grandits6Gerhard F. Ecker7Roderick L. Beijersbergen8Peter Bouwman9Sylvia E. Le Dévédec10Bob van de Water11Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden UniversityDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden UniversityDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden UniversityDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden UniversityDivision of Molecular Carcinogenesis, The NKI Robotics and Screening Center, The Netherlands Cancer InstituteDivision of Molecular Carcinogenesis, The NKI Robotics and Screening Center, The Netherlands Cancer InstituteDepartment of Pharmaceutical Sciences, University of ViennaDepartment of Pharmaceutical Sciences, University of ViennaDivision of Molecular Carcinogenesis, The NKI Robotics and Screening Center, The Netherlands Cancer InstituteDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden UniversityDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden UniversityDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden UniversityAbstract Background Triple-negative breast cancer (TNBC) is a subtype of breast cancer with limited treatment options and poor clinical prognosis. Inhibitors of transcriptional CDKs are currently under thorough investigation for application in the treatment of multiple cancer types, including breast cancer. These studies have raised interest in combining these inhibitors, including CDK12/13 inhibitor THZ531, with a variety of other anti-cancer agents. However, the full scope of these potential synergistic interactions of transcriptional CDK inhibitors with kinase inhibitors has not been systematically investigated. Moreover, the mechanisms behind these previously described synergistic interactions remain largely elusive. Methods Kinase inhibitor combination screenings were performed to identify kinase inhibitors that synergize with CDK7 inhibitor THZ1 and CDK12/13 inhibitor THZ531 in TNBC cell lines. CRISPR-Cas9 knockout screening and transcriptomic evaluation of resistant versus sensitive cell lines were performed to identify genes critical for THZ531 resistance. RNA sequencing analysis after treatment with individual and combined synergistic treatments was performed to gain further insights into the mechanism of this synergy. Kinase inhibitor screening in combination with visualization of ABCG2-substrate pheophorbide A was used to identify kinase inhibitors that inhibit ABCG2. Multiple transcriptional CDK inhibitors were evaluated to extend the significance of the found mechanism to other transcriptional CDK inhibitors. Results We show that a very high number of tyrosine kinase inhibitors synergize with the CDK12/13 inhibitor THZ531. Yet, we identified the multidrug transporter ABCG2 as key determinant of THZ531 resistance in TNBC cells. Mechanistically, we demonstrate that most synergistic kinase inhibitors block ABCG2 function, thereby sensitizing cells to transcriptional CDK inhibitors, including THZ531. Accordingly, these kinase inhibitors potentiate the effects of THZ531, disrupting gene expression and increasing intronic polyadenylation. Conclusion Overall, this study demonstrates the critical role of ABCG2 in limiting the efficacy of transcriptional CDK inhibitors and identifies multiple kinase inhibitors that disrupt ABCG2 transporter function and thereby synergize with these CDK inhibitors. These findings therefore further facilitate the development of new (combination) therapies targeting transcriptional CDKs and highlight the importance of evaluating the role of ABC transporters in synergistic drug–drug interactions in general.https://doi.org/10.1186/s13058-023-01648-xTriple-negative breast cancerTranscriptional cyclin-dependent kinasesCDK12/13THZ531ABCG2Drug resistance |
| spellingShingle | Vera E. van der Noord Wanda van der Stel Gijs Louwerens Danielle Verhoeven Hendrik J. Kuiken Cor Lieftink Melanie Grandits Gerhard F. Ecker Roderick L. Beijersbergen Peter Bouwman Sylvia E. Le Dévédec Bob van de Water Systematic screening identifies ABCG2 as critical factor underlying synergy of kinase inhibitors with transcriptional CDK inhibitors Breast Cancer Research Triple-negative breast cancer Transcriptional cyclin-dependent kinases CDK12/13 THZ531 ABCG2 Drug resistance |
| title | Systematic screening identifies ABCG2 as critical factor underlying synergy of kinase inhibitors with transcriptional CDK inhibitors |
| title_full | Systematic screening identifies ABCG2 as critical factor underlying synergy of kinase inhibitors with transcriptional CDK inhibitors |
| title_fullStr | Systematic screening identifies ABCG2 as critical factor underlying synergy of kinase inhibitors with transcriptional CDK inhibitors |
| title_full_unstemmed | Systematic screening identifies ABCG2 as critical factor underlying synergy of kinase inhibitors with transcriptional CDK inhibitors |
| title_short | Systematic screening identifies ABCG2 as critical factor underlying synergy of kinase inhibitors with transcriptional CDK inhibitors |
| title_sort | systematic screening identifies abcg2 as critical factor underlying synergy of kinase inhibitors with transcriptional cdk inhibitors |
| topic | Triple-negative breast cancer Transcriptional cyclin-dependent kinases CDK12/13 THZ531 ABCG2 Drug resistance |
| url | https://doi.org/10.1186/s13058-023-01648-x |
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