A phenotypic screening approach to target p60AmotL2-expressing invasive cancer cells
Abstract Background Tumor cells have the ability to invade and form small clusters that protrude into adjacent tissues, a phenomenon that is frequently observed at the periphery of a tumor as it expands into healthy tissues. The presence of these clusters is linked to poor prognosis and has proven c...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-04-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13046-024-03031-w |
| _version_ | 1797208981014315008 |
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| author | Pedro Fonseca Weiyingqi Cui Nona Struyf Le Tong Ayushi Chaurasiya Felipe Casagrande Honglei Zhao Dinura Fernando Xinsong Chen Nicholas P. Tobin Brinton Seashore-Ludlow Andreas Lundqvist Johan Hartman Anita Göndör Päivi Östling Lars Holmgren |
| author_facet | Pedro Fonseca Weiyingqi Cui Nona Struyf Le Tong Ayushi Chaurasiya Felipe Casagrande Honglei Zhao Dinura Fernando Xinsong Chen Nicholas P. Tobin Brinton Seashore-Ludlow Andreas Lundqvist Johan Hartman Anita Göndör Päivi Östling Lars Holmgren |
| author_sort | Pedro Fonseca |
| collection | DOAJ |
| description | Abstract Background Tumor cells have the ability to invade and form small clusters that protrude into adjacent tissues, a phenomenon that is frequently observed at the periphery of a tumor as it expands into healthy tissues. The presence of these clusters is linked to poor prognosis and has proven challenging to treat using conventional therapies. We previously reported that p60AmotL2 expression is localized to invasive colon and breast cancer cells. In vitro, p60AmotL2 promotes epithelial cell invasion by negatively impacting E-cadherin/AmotL2-related mechanotransduction. Methods Using epithelial cells transfected with inducible p60AmotL2, we employed a phenotypic drug screening approach to find compounds that specifically target invasive cells. The phenotypic screen was performed by treating cells for 72 h with a library of compounds with known antitumor activities in a dose-dependent manner. After assessing cell viability using CellTiter-Glo, drug sensitivity scores for each compound were calculated. Candidate hit compounds with a higher drug sensitivity score for p60AmotL2-expressing cells were then validated on lung and colon cell models, both in 2D and in 3D, and on colon cancer patient-derived organoids. Nascent RNA sequencing was performed after BET inhibition to analyse BET-dependent pathways in p60AmotL2-expressing cells. Results We identified 60 compounds that selectively targeted p60AmotL2-expressing cells. Intriguingly, these compounds were classified into two major categories: Epidermal Growth Factor Receptor (EGFR) inhibitors and Bromodomain and Extra-Terminal motif (BET) inhibitors. The latter consistently demonstrated antitumor activity in human cancer cell models, as well as in organoids derived from colon cancer patients. BET inhibition led to a shift towards the upregulation of pro-apoptotic pathways specifically in p60AmotL2-expressing cells. Conclusions BET inhibitors specifically target p60AmotL2-expressing invasive cancer cells, likely by exploiting differences in chromatin accessibility, leading to cell death. Additionally, our findings support the use of this phenotypic strategy to discover novel compounds that can exploit vulnerabilities and specifically target invasive cancer cells. |
| first_indexed | 2024-04-24T09:47:26Z |
| format | Article |
| id | doaj.art-33dfb61ce30b4942b583460a044384b6 |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-04-24T09:47:26Z |
| publishDate | 2024-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-33dfb61ce30b4942b583460a044384b62024-04-14T11:32:45ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662024-04-0143112310.1186/s13046-024-03031-wA phenotypic screening approach to target p60AmotL2-expressing invasive cancer cellsPedro Fonseca0Weiyingqi Cui1Nona Struyf2Le Tong3Ayushi Chaurasiya4Felipe Casagrande5Honglei Zhao6Dinura Fernando7Xinsong Chen8Nicholas P. Tobin9Brinton Seashore-Ludlow10Andreas Lundqvist11Johan Hartman12Anita Göndör13Päivi Östling14Lars Holmgren15Department of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetDepartment of Oncology and Pathology, Karolinska InstitutetAbstract Background Tumor cells have the ability to invade and form small clusters that protrude into adjacent tissues, a phenomenon that is frequently observed at the periphery of a tumor as it expands into healthy tissues. The presence of these clusters is linked to poor prognosis and has proven challenging to treat using conventional therapies. We previously reported that p60AmotL2 expression is localized to invasive colon and breast cancer cells. In vitro, p60AmotL2 promotes epithelial cell invasion by negatively impacting E-cadherin/AmotL2-related mechanotransduction. Methods Using epithelial cells transfected with inducible p60AmotL2, we employed a phenotypic drug screening approach to find compounds that specifically target invasive cells. The phenotypic screen was performed by treating cells for 72 h with a library of compounds with known antitumor activities in a dose-dependent manner. After assessing cell viability using CellTiter-Glo, drug sensitivity scores for each compound were calculated. Candidate hit compounds with a higher drug sensitivity score for p60AmotL2-expressing cells were then validated on lung and colon cell models, both in 2D and in 3D, and on colon cancer patient-derived organoids. Nascent RNA sequencing was performed after BET inhibition to analyse BET-dependent pathways in p60AmotL2-expressing cells. Results We identified 60 compounds that selectively targeted p60AmotL2-expressing cells. Intriguingly, these compounds were classified into two major categories: Epidermal Growth Factor Receptor (EGFR) inhibitors and Bromodomain and Extra-Terminal motif (BET) inhibitors. The latter consistently demonstrated antitumor activity in human cancer cell models, as well as in organoids derived from colon cancer patients. BET inhibition led to a shift towards the upregulation of pro-apoptotic pathways specifically in p60AmotL2-expressing cells. Conclusions BET inhibitors specifically target p60AmotL2-expressing invasive cancer cells, likely by exploiting differences in chromatin accessibility, leading to cell death. Additionally, our findings support the use of this phenotypic strategy to discover novel compounds that can exploit vulnerabilities and specifically target invasive cancer cells.https://doi.org/10.1186/s13046-024-03031-wCancerCancer invasionMetastasisMechanotransductionPhenotypic drug screeningPatient-derived organoids |
| spellingShingle | Pedro Fonseca Weiyingqi Cui Nona Struyf Le Tong Ayushi Chaurasiya Felipe Casagrande Honglei Zhao Dinura Fernando Xinsong Chen Nicholas P. Tobin Brinton Seashore-Ludlow Andreas Lundqvist Johan Hartman Anita Göndör Päivi Östling Lars Holmgren A phenotypic screening approach to target p60AmotL2-expressing invasive cancer cells Journal of Experimental & Clinical Cancer Research Cancer Cancer invasion Metastasis Mechanotransduction Phenotypic drug screening Patient-derived organoids |
| title | A phenotypic screening approach to target p60AmotL2-expressing invasive cancer cells |
| title_full | A phenotypic screening approach to target p60AmotL2-expressing invasive cancer cells |
| title_fullStr | A phenotypic screening approach to target p60AmotL2-expressing invasive cancer cells |
| title_full_unstemmed | A phenotypic screening approach to target p60AmotL2-expressing invasive cancer cells |
| title_short | A phenotypic screening approach to target p60AmotL2-expressing invasive cancer cells |
| title_sort | phenotypic screening approach to target p60amotl2 expressing invasive cancer cells |
| topic | Cancer Cancer invasion Metastasis Mechanotransduction Phenotypic drug screening Patient-derived organoids |
| url | https://doi.org/10.1186/s13046-024-03031-w |
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