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The MDM2 Inhibitor Navtemadlin Arrests Mouse Melanoma Growth In Vivo and Potentiates Radiotherapy
The tumor suppressor protein p53 is mutated in close to 50% of human tumors and is dysregulated in many others, for instance by silencing or loss of p14(ARF). Under steady-state conditions, the two E3 ligases MDM2/MDM4 interact with and inhibit the transcriptional activity of p53. Inhibition of p53–...
Autores principales: | , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Association for Cancer Research
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10010373/ https://www.ncbi.nlm.nih.gov/pubmed/36922937 http://dx.doi.org/10.1158/2767-9764.CRC-22-0053 |
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author | Ingelshed, Katrine Spiegelberg, Diana Kannan, Pavitra Påvénius, Linnéa Hacheney, Jessica Jiang, Long Eisinger, Silke Lianoudaki, Danai Lama, Dilraj Castillo, Francisca Bosdotter, Cecilia Kretzschmar, Warren W. Al-Radi, Omayma Fritz, Nicolas Villablanca, Eduardo J. Karlsson, Mikael C. I. Wermeling, Fredrik Nestor, Marika Lane, David P. Sedimbi, Saikiran K. |
author_facet | Ingelshed, Katrine Spiegelberg, Diana Kannan, Pavitra Påvénius, Linnéa Hacheney, Jessica Jiang, Long Eisinger, Silke Lianoudaki, Danai Lama, Dilraj Castillo, Francisca Bosdotter, Cecilia Kretzschmar, Warren W. Al-Radi, Omayma Fritz, Nicolas Villablanca, Eduardo J. Karlsson, Mikael C. I. Wermeling, Fredrik Nestor, Marika Lane, David P. Sedimbi, Saikiran K. |
author_sort | Ingelshed, Katrine |
collection | PubMed |
description | The tumor suppressor protein p53 is mutated in close to 50% of human tumors and is dysregulated in many others, for instance by silencing or loss of p14(ARF). Under steady-state conditions, the two E3 ligases MDM2/MDM4 interact with and inhibit the transcriptional activity of p53. Inhibition of p53–MDM2/4 interaction to reactivate p53 in tumors with wild-type (WT) p53 has therefore been considered a therapeutic strategy. Moreover, studies indicate that p53 reactivation may synergize with radiation and increase tumor immunogenicity. In vivo studies of most MDM2 inhibitors have utilized immunodeficient xenograft mouse models, preventing detailed studies of action of these molecules on the immune response. The mouse melanoma cell line B16-F10 carries functional, WT p53 but does not express the MDM2 regulator p19(ARF). In this study, we tested a p53-MDM2 protein–protein interaction inhibitor, the small molecule Navtemadlin, which is currently being tested in phase II clinical trials. Using mass spectrometry–based proteomics and imaging flow cytometry, we identified specific protein expression patterns following Navtemadlin treatment of B16-F10 melanoma cells compared with their p53 CRISPR-inactivated control cells. In vitro, Navtemadlin induced a significant, p53-dependent, growth arrest but little apoptosis in B16-F10 cells. When combined with radiotherapy, Navtemadlin showed synergistic effects and increased apoptosis. In vivo, Navtemadlin treatment significantly reduced the growth of B16-F10 melanoma cells implanted in C57Bl/6 mice. Our data highlight the utility of a syngeneic B16-F10 p53(+/+) mouse melanoma model for assessing existing and novel p53-MDM2/MDM4 inhibitors and in identifying new combination therapies that can efficiently eliminate tumors in vivo. SIGNIFICANCE: The MDM2 inhibitor Navtemadlin arrests mouse tumor growth and potentiates radiotherapy. Our results support a threshold model for apoptosis induction that requires a high, prolonged p53 signaling for cancer cells to become apoptotic. |
format | Online Article Text |
id | pubmed-10010373 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Association for Cancer Research |
record_format | MEDLINE/PubMed |
spelling | pubmed-100103732023-03-14 The MDM2 Inhibitor Navtemadlin Arrests Mouse Melanoma Growth In Vivo and Potentiates Radiotherapy Ingelshed, Katrine Spiegelberg, Diana Kannan, Pavitra Påvénius, Linnéa Hacheney, Jessica Jiang, Long Eisinger, Silke Lianoudaki, Danai Lama, Dilraj Castillo, Francisca Bosdotter, Cecilia Kretzschmar, Warren W. Al-Radi, Omayma Fritz, Nicolas Villablanca, Eduardo J. Karlsson, Mikael C. I. Wermeling, Fredrik Nestor, Marika Lane, David P. Sedimbi, Saikiran K. Cancer Res Commun Research Article The tumor suppressor protein p53 is mutated in close to 50% of human tumors and is dysregulated in many others, for instance by silencing or loss of p14(ARF). Under steady-state conditions, the two E3 ligases MDM2/MDM4 interact with and inhibit the transcriptional activity of p53. Inhibition of p53–MDM2/4 interaction to reactivate p53 in tumors with wild-type (WT) p53 has therefore been considered a therapeutic strategy. Moreover, studies indicate that p53 reactivation may synergize with radiation and increase tumor immunogenicity. In vivo studies of most MDM2 inhibitors have utilized immunodeficient xenograft mouse models, preventing detailed studies of action of these molecules on the immune response. The mouse melanoma cell line B16-F10 carries functional, WT p53 but does not express the MDM2 regulator p19(ARF). In this study, we tested a p53-MDM2 protein–protein interaction inhibitor, the small molecule Navtemadlin, which is currently being tested in phase II clinical trials. Using mass spectrometry–based proteomics and imaging flow cytometry, we identified specific protein expression patterns following Navtemadlin treatment of B16-F10 melanoma cells compared with their p53 CRISPR-inactivated control cells. In vitro, Navtemadlin induced a significant, p53-dependent, growth arrest but little apoptosis in B16-F10 cells. When combined with radiotherapy, Navtemadlin showed synergistic effects and increased apoptosis. In vivo, Navtemadlin treatment significantly reduced the growth of B16-F10 melanoma cells implanted in C57Bl/6 mice. Our data highlight the utility of a syngeneic B16-F10 p53(+/+) mouse melanoma model for assessing existing and novel p53-MDM2/MDM4 inhibitors and in identifying new combination therapies that can efficiently eliminate tumors in vivo. SIGNIFICANCE: The MDM2 inhibitor Navtemadlin arrests mouse tumor growth and potentiates radiotherapy. Our results support a threshold model for apoptosis induction that requires a high, prolonged p53 signaling for cancer cells to become apoptotic. American Association for Cancer Research 2022-09-28 /pmc/articles/PMC10010373/ /pubmed/36922937 http://dx.doi.org/10.1158/2767-9764.CRC-22-0053 Text en © 2022 The Authors; Published by the American Association for Cancer Research https://creativecommons.org/licenses/by/4.0/This open access article is distributed under the Creative Commons Attribution 4.0 International (CC BY 4.0) license. |
spellingShingle | Research Article Ingelshed, Katrine Spiegelberg, Diana Kannan, Pavitra Påvénius, Linnéa Hacheney, Jessica Jiang, Long Eisinger, Silke Lianoudaki, Danai Lama, Dilraj Castillo, Francisca Bosdotter, Cecilia Kretzschmar, Warren W. Al-Radi, Omayma Fritz, Nicolas Villablanca, Eduardo J. Karlsson, Mikael C. I. Wermeling, Fredrik Nestor, Marika Lane, David P. Sedimbi, Saikiran K. The MDM2 Inhibitor Navtemadlin Arrests Mouse Melanoma Growth In Vivo and Potentiates Radiotherapy |
title | The MDM2 Inhibitor Navtemadlin Arrests Mouse Melanoma Growth In Vivo and Potentiates Radiotherapy |
title_full | The MDM2 Inhibitor Navtemadlin Arrests Mouse Melanoma Growth In Vivo and Potentiates Radiotherapy |
title_fullStr | The MDM2 Inhibitor Navtemadlin Arrests Mouse Melanoma Growth In Vivo and Potentiates Radiotherapy |
title_full_unstemmed | The MDM2 Inhibitor Navtemadlin Arrests Mouse Melanoma Growth In Vivo and Potentiates Radiotherapy |
title_short | The MDM2 Inhibitor Navtemadlin Arrests Mouse Melanoma Growth In Vivo and Potentiates Radiotherapy |
title_sort | mdm2 inhibitor navtemadlin arrests mouse melanoma growth in vivo and potentiates radiotherapy |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10010373/ https://www.ncbi.nlm.nih.gov/pubmed/36922937 http://dx.doi.org/10.1158/2767-9764.CRC-22-0053 |
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