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RNA interference screens discover proteases as synthetic lethal partners of PI3K inhibition in breast cancer cells
Rationale: PI3K/mTOR signaling is frequently upregulated in breast cancer making inhibitors of this pathway highly promising anticancer drugs. However, PI3K-inhibitors have a low therapeutic index. Therefore, finding novel combinatory treatment options represents an important step towards clinical i...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Ivyspring International Publisher
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9169373/ https://www.ncbi.nlm.nih.gov/pubmed/35673573 http://dx.doi.org/10.7150/thno.68299 |
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author | Hölzen, Lena Mitschke, Jan Schönichen, Claudia Hess, Maria Elena Ehrenfeld, Sophia Boerries, Melanie Miething, Cornelius Brummer, Tilman Reinheckel, Thomas |
author_facet | Hölzen, Lena Mitschke, Jan Schönichen, Claudia Hess, Maria Elena Ehrenfeld, Sophia Boerries, Melanie Miething, Cornelius Brummer, Tilman Reinheckel, Thomas |
author_sort | Hölzen, Lena |
collection | PubMed |
description | Rationale: PI3K/mTOR signaling is frequently upregulated in breast cancer making inhibitors of this pathway highly promising anticancer drugs. However, PI3K-inhibitors have a low therapeutic index. Therefore, finding novel combinatory treatment options represents an important step towards clinical implementation of PI3K pathway inhibition in breast cancer therapy. Here, we propose proteases as potential synergistic partners with simultaneous PI3K inhibition in breast cancer cells. Methods: We performed mRNA expression studies and unbiased functional genetic synthetic lethality screens by a miR-E based knockdown system targeting all genome-encoded proteases, i.e. the degradome of breast cancer cells. Importantly theses RNA interference screens were done in combination with two PI3K pathway inhibitors. Protease hits were validated in human and murine breast cancer cell lines as well as in non-cancerous cells by viability and growth assays. Results: The degradome-wide genetic screens identified 181 proteases that influenced susceptibility of murine breast cancer cells to low dose PI3K inhibition. Employing independently generated inducible knockdown cell lines we validated 12 protease hits in breast cancer cells. In line with the known tumor promoting function of these proteases we demonstrated Usp7 and Metap2 to be important for murine and human breast cancer cell growth and discovered a role for Metap1 in this context. Most importantly, we demonstrated that Usp7, Metap1 or Metap2 knockdown combined with simultaneous PI3K inhibition resulted in synergistic impairment of murine and human breast cancer cell growth Conclusion: We successfully established proteases as combinatory targets with PI3K inhibition in human and murine breast cancer cells. Usp7, Metap1 and Metap2 are synthetic lethal partners of simultaneous protease/PI3K inhibition, which may refine future breast cancer therapy. |
format | Online Article Text |
id | pubmed-9169373 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Ivyspring International Publisher |
record_format | MEDLINE/PubMed |
spelling | pubmed-91693732022-06-06 RNA interference screens discover proteases as synthetic lethal partners of PI3K inhibition in breast cancer cells Hölzen, Lena Mitschke, Jan Schönichen, Claudia Hess, Maria Elena Ehrenfeld, Sophia Boerries, Melanie Miething, Cornelius Brummer, Tilman Reinheckel, Thomas Theranostics Research Paper Rationale: PI3K/mTOR signaling is frequently upregulated in breast cancer making inhibitors of this pathway highly promising anticancer drugs. However, PI3K-inhibitors have a low therapeutic index. Therefore, finding novel combinatory treatment options represents an important step towards clinical implementation of PI3K pathway inhibition in breast cancer therapy. Here, we propose proteases as potential synergistic partners with simultaneous PI3K inhibition in breast cancer cells. Methods: We performed mRNA expression studies and unbiased functional genetic synthetic lethality screens by a miR-E based knockdown system targeting all genome-encoded proteases, i.e. the degradome of breast cancer cells. Importantly theses RNA interference screens were done in combination with two PI3K pathway inhibitors. Protease hits were validated in human and murine breast cancer cell lines as well as in non-cancerous cells by viability and growth assays. Results: The degradome-wide genetic screens identified 181 proteases that influenced susceptibility of murine breast cancer cells to low dose PI3K inhibition. Employing independently generated inducible knockdown cell lines we validated 12 protease hits in breast cancer cells. In line with the known tumor promoting function of these proteases we demonstrated Usp7 and Metap2 to be important for murine and human breast cancer cell growth and discovered a role for Metap1 in this context. Most importantly, we demonstrated that Usp7, Metap1 or Metap2 knockdown combined with simultaneous PI3K inhibition resulted in synergistic impairment of murine and human breast cancer cell growth Conclusion: We successfully established proteases as combinatory targets with PI3K inhibition in human and murine breast cancer cells. Usp7, Metap1 and Metap2 are synthetic lethal partners of simultaneous protease/PI3K inhibition, which may refine future breast cancer therapy. Ivyspring International Publisher 2022-05-16 /pmc/articles/PMC9169373/ /pubmed/35673573 http://dx.doi.org/10.7150/thno.68299 Text en © The author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions. |
spellingShingle | Research Paper Hölzen, Lena Mitschke, Jan Schönichen, Claudia Hess, Maria Elena Ehrenfeld, Sophia Boerries, Melanie Miething, Cornelius Brummer, Tilman Reinheckel, Thomas RNA interference screens discover proteases as synthetic lethal partners of PI3K inhibition in breast cancer cells |
title | RNA interference screens discover proteases as synthetic lethal partners of PI3K inhibition in breast cancer cells |
title_full | RNA interference screens discover proteases as synthetic lethal partners of PI3K inhibition in breast cancer cells |
title_fullStr | RNA interference screens discover proteases as synthetic lethal partners of PI3K inhibition in breast cancer cells |
title_full_unstemmed | RNA interference screens discover proteases as synthetic lethal partners of PI3K inhibition in breast cancer cells |
title_short | RNA interference screens discover proteases as synthetic lethal partners of PI3K inhibition in breast cancer cells |
title_sort | rna interference screens discover proteases as synthetic lethal partners of pi3k inhibition in breast cancer cells |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9169373/ https://www.ncbi.nlm.nih.gov/pubmed/35673573 http://dx.doi.org/10.7150/thno.68299 |
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