Wee1 Inhibition Enhances the Anti-Tumor Effects of Capecitabine in Preclinical Models of Triple-Negative Breast Cancer
Triple-negative breast cancer (TNBC) is an aggressive subtype defined by lack of hormone receptor expression and non-amplified HER2. Adavosertib (AZD1775) is a potent, small-molecule, ATP-competitive inhibitor of the Wee1 kinase that potentiates the activity of many DNA-damaging chemotherapeutics an...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-03-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/12/3/719 |
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| author | Todd M. Pitts Dennis M. Simmons Stacey M. Bagby Sarah J. Hartman Betelehem W. Yacob Brian Gittleman John J. Tentler Diana Cittelly D. Ryan Ormond Wells A. Messersmith S. Gail Eckhardt Jennifer R. Diamond |
| author_facet | Todd M. Pitts Dennis M. Simmons Stacey M. Bagby Sarah J. Hartman Betelehem W. Yacob Brian Gittleman John J. Tentler Diana Cittelly D. Ryan Ormond Wells A. Messersmith S. Gail Eckhardt Jennifer R. Diamond |
| author_sort | Todd M. Pitts |
| collection | DOAJ |
| description | Triple-negative breast cancer (TNBC) is an aggressive subtype defined by lack of hormone receptor expression and non-amplified HER2. Adavosertib (AZD1775) is a potent, small-molecule, ATP-competitive inhibitor of the Wee1 kinase that potentiates the activity of many DNA-damaging chemotherapeutics and is currently in clinical development for multiple indications. The purpose of this study was to investigate the combination of AZD1775 and capecitabine/5FU in preclinical TNBC models. TNBC cell lines were treated with AZD1775 and 5FU and cellular proliferation was assessed in real-time using IncuCyte<sup>®</sup> Live Cell Analysis. Apoptosis was assessed via the Caspase-Glo 3/7 assay system. Western blotting was used to assess changes in expression of downstream effectors. TNBC patient-derived xenograft (PDX) models were treated with AZD1775, capecitabine, or the combination and assessed for tumor growth inhibition. From the initial PDX screen, two of the four TNBC PDX models demonstrated a better response in the combination treatment than either of the single agents. As confirmation, two PDX models were expanded for statistical comparison. Both PDX models demonstrated a significant growth inhibition in the combination versus either of the single agents. (TNBC012, <i>p</i> < 0.05 combo vs. adavosertib or capecitabine, TNBC013, <i>p</i> < 0.01 combo vs. adavosertib or capecitabine.) An enhanced anti-proliferative effect was observed in the adavosertib/5FU combination treatment as measured by live cell analysis. An increase in apoptosis was observed in two of the four cell lines in the combination when compared to single-agent treatment. Treatment with adavosertib as a single agent resulted in a decrease in p-CDC2 in a dose-dependent manner that was also observed in the combination treatment. An increase in γH2AX in two of the four cell lines tested was also observed. No significant changes were observed in Bcl-xL following treatment in any of the cell lines. The combination of adavosertib and capecitabine/5FU demonstrated enhanced combination effects both in vitro and in vivo in preclinical models of TNBC. These results support the clinical investigation of this combination in patients with TNBC, including those with brain metastasis given the CNS penetration of both agents. |
| first_indexed | 2024-03-12T19:59:02Z |
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| id | doaj.art-98303b9316db4c7cb0e1e6133ffcdc79 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-12T19:59:02Z |
| publishDate | 2020-03-01 |
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| series | Cancers |
| spelling | doaj.art-98303b9316db4c7cb0e1e6133ffcdc792023-08-02T02:34:02ZengMDPI AGCancers2072-66942020-03-0112371910.3390/cancers12030719cancers12030719Wee1 Inhibition Enhances the Anti-Tumor Effects of Capecitabine in Preclinical Models of Triple-Negative Breast CancerTodd M. Pitts0Dennis M. Simmons1Stacey M. Bagby2Sarah J. Hartman3Betelehem W. Yacob4Brian Gittleman5John J. Tentler6Diana Cittelly7D. Ryan Ormond8Wells A. Messersmith9S. Gail Eckhardt10Jennifer R. Diamond11Division of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADivision of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADivision of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADivision of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADivision of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADivision of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADivision of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADepartment of Pathology, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADepartment of Neurosurgery, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADivision of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USADell Medical School, Department of Oncology, The University of Texas Austin, 1701 Trinity Street, Austin, TX 78712, USADivision of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO 80045, USATriple-negative breast cancer (TNBC) is an aggressive subtype defined by lack of hormone receptor expression and non-amplified HER2. Adavosertib (AZD1775) is a potent, small-molecule, ATP-competitive inhibitor of the Wee1 kinase that potentiates the activity of many DNA-damaging chemotherapeutics and is currently in clinical development for multiple indications. The purpose of this study was to investigate the combination of AZD1775 and capecitabine/5FU in preclinical TNBC models. TNBC cell lines were treated with AZD1775 and 5FU and cellular proliferation was assessed in real-time using IncuCyte<sup>®</sup> Live Cell Analysis. Apoptosis was assessed via the Caspase-Glo 3/7 assay system. Western blotting was used to assess changes in expression of downstream effectors. TNBC patient-derived xenograft (PDX) models were treated with AZD1775, capecitabine, or the combination and assessed for tumor growth inhibition. From the initial PDX screen, two of the four TNBC PDX models demonstrated a better response in the combination treatment than either of the single agents. As confirmation, two PDX models were expanded for statistical comparison. Both PDX models demonstrated a significant growth inhibition in the combination versus either of the single agents. (TNBC012, <i>p</i> < 0.05 combo vs. adavosertib or capecitabine, TNBC013, <i>p</i> < 0.01 combo vs. adavosertib or capecitabine.) An enhanced anti-proliferative effect was observed in the adavosertib/5FU combination treatment as measured by live cell analysis. An increase in apoptosis was observed in two of the four cell lines in the combination when compared to single-agent treatment. Treatment with adavosertib as a single agent resulted in a decrease in p-CDC2 in a dose-dependent manner that was also observed in the combination treatment. An increase in γH2AX in two of the four cell lines tested was also observed. No significant changes were observed in Bcl-xL following treatment in any of the cell lines. The combination of adavosertib and capecitabine/5FU demonstrated enhanced combination effects both in vitro and in vivo in preclinical models of TNBC. These results support the clinical investigation of this combination in patients with TNBC, including those with brain metastasis given the CNS penetration of both agents.https://www.mdpi.com/2072-6694/12/3/719triple-negative breast cancerwee15-fudna damage |
| spellingShingle | Todd M. Pitts Dennis M. Simmons Stacey M. Bagby Sarah J. Hartman Betelehem W. Yacob Brian Gittleman John J. Tentler Diana Cittelly D. Ryan Ormond Wells A. Messersmith S. Gail Eckhardt Jennifer R. Diamond Wee1 Inhibition Enhances the Anti-Tumor Effects of Capecitabine in Preclinical Models of Triple-Negative Breast Cancer Cancers triple-negative breast cancer wee1 5-fu dna damage |
| title | Wee1 Inhibition Enhances the Anti-Tumor Effects of Capecitabine in Preclinical Models of Triple-Negative Breast Cancer |
| title_full | Wee1 Inhibition Enhances the Anti-Tumor Effects of Capecitabine in Preclinical Models of Triple-Negative Breast Cancer |
| title_fullStr | Wee1 Inhibition Enhances the Anti-Tumor Effects of Capecitabine in Preclinical Models of Triple-Negative Breast Cancer |
| title_full_unstemmed | Wee1 Inhibition Enhances the Anti-Tumor Effects of Capecitabine in Preclinical Models of Triple-Negative Breast Cancer |
| title_short | Wee1 Inhibition Enhances the Anti-Tumor Effects of Capecitabine in Preclinical Models of Triple-Negative Breast Cancer |
| title_sort | wee1 inhibition enhances the anti tumor effects of capecitabine in preclinical models of triple negative breast cancer |
| topic | triple-negative breast cancer wee1 5-fu dna damage |
| url | https://www.mdpi.com/2072-6694/12/3/719 |
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