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Degradome-focused RNA interference screens to identify proteases important for breast cancer cell growth

Proteases are known to promote or impair breast cancer progression and metastasis. However, while a small number of the 588 human and 672 murine protease genes have been extensively studied, others were neglected. For an unbiased functional analysis of all genome-encoded proteases, i.e., the degrado...

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Autores principales: Hölzen, Lena, Syré, Kerstin, Mitschke, Jan, Brummer, Tilman, Miething, Cornelius, Reinheckel, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9598039/
https://www.ncbi.nlm.nih.gov/pubmed/36313646
http://dx.doi.org/10.3389/fonc.2022.960109
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author Hölzen, Lena
Syré, Kerstin
Mitschke, Jan
Brummer, Tilman
Miething, Cornelius
Reinheckel, Thomas
author_facet Hölzen, Lena
Syré, Kerstin
Mitschke, Jan
Brummer, Tilman
Miething, Cornelius
Reinheckel, Thomas
author_sort Hölzen, Lena
collection PubMed
description Proteases are known to promote or impair breast cancer progression and metastasis. However, while a small number of the 588 human and 672 murine protease genes have been extensively studied, others were neglected. For an unbiased functional analysis of all genome-encoded proteases, i.e., the degradome, in breast cancer cell growth, we applied an inducible RNA interference library for protease-focused genetic screens. Importantly, these functional screens were performed in two phenotypically different murine breast cancer cell lines, including one stem cell-like cell line that showed phenotypic plasticity under changed nutrient and oxygen availability. Our unbiased genetic screens identified 252 protease genes involved in breast cancer cell growth that were further restricted to 100 hits by a selection process. Many of those hits were supported by literature, but some proteases were novel in their functional link to breast cancer. Interestingly, we discovered that the environmental conditions influence the degree of breast cancer cell dependency on certain proteases. For example, breast cancer stem cell-like cells were less susceptible to depletion of several mitochondrial proteases in hypoxic conditions. From the 100 hits, nine proteases were functionally validated in murine breast cancer cell lines using individual knockdown constructs, highlighting the high reliability of our screens. Specifically, we focused on mitochondrial processing peptidase (MPP) subunits alpha (Pmpca) and beta (Pmpcb) and discovered that MPP depletion led to a disadvantage in cell growth, which was linked to mitochondrial dysfunction.
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spelling pubmed-95980392022-10-27 Degradome-focused RNA interference screens to identify proteases important for breast cancer cell growth Hölzen, Lena Syré, Kerstin Mitschke, Jan Brummer, Tilman Miething, Cornelius Reinheckel, Thomas Front Oncol Oncology Proteases are known to promote or impair breast cancer progression and metastasis. However, while a small number of the 588 human and 672 murine protease genes have been extensively studied, others were neglected. For an unbiased functional analysis of all genome-encoded proteases, i.e., the degradome, in breast cancer cell growth, we applied an inducible RNA interference library for protease-focused genetic screens. Importantly, these functional screens were performed in two phenotypically different murine breast cancer cell lines, including one stem cell-like cell line that showed phenotypic plasticity under changed nutrient and oxygen availability. Our unbiased genetic screens identified 252 protease genes involved in breast cancer cell growth that were further restricted to 100 hits by a selection process. Many of those hits were supported by literature, but some proteases were novel in their functional link to breast cancer. Interestingly, we discovered that the environmental conditions influence the degree of breast cancer cell dependency on certain proteases. For example, breast cancer stem cell-like cells were less susceptible to depletion of several mitochondrial proteases in hypoxic conditions. From the 100 hits, nine proteases were functionally validated in murine breast cancer cell lines using individual knockdown constructs, highlighting the high reliability of our screens. Specifically, we focused on mitochondrial processing peptidase (MPP) subunits alpha (Pmpca) and beta (Pmpcb) and discovered that MPP depletion led to a disadvantage in cell growth, which was linked to mitochondrial dysfunction. Frontiers Media S.A. 2022-10-12 /pmc/articles/PMC9598039/ /pubmed/36313646 http://dx.doi.org/10.3389/fonc.2022.960109 Text en Copyright © 2022 Hölzen, Syré, Mitschke, Brummer, Miething and Reinheckel https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Oncology
Hölzen, Lena
Syré, Kerstin
Mitschke, Jan
Brummer, Tilman
Miething, Cornelius
Reinheckel, Thomas
Degradome-focused RNA interference screens to identify proteases important for breast cancer cell growth
title Degradome-focused RNA interference screens to identify proteases important for breast cancer cell growth
title_full Degradome-focused RNA interference screens to identify proteases important for breast cancer cell growth
title_fullStr Degradome-focused RNA interference screens to identify proteases important for breast cancer cell growth
title_full_unstemmed Degradome-focused RNA interference screens to identify proteases important for breast cancer cell growth
title_short Degradome-focused RNA interference screens to identify proteases important for breast cancer cell growth
title_sort degradome-focused rna interference screens to identify proteases important for breast cancer cell growth
topic Oncology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9598039/
https://www.ncbi.nlm.nih.gov/pubmed/36313646
http://dx.doi.org/10.3389/fonc.2022.960109
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