Identification of Src as a Therapeutic Target in Oesophageal Adenocarcinoma through Functional Genomic and High-Throughput Drug Screening Approaches
Drug resistance limits the effectiveness of oesophageal adenocarcinoma (OAC) chemotherapies, leading to a poor prognosis for this disease. Elucidation of the underlying resistance mechanisms is key to enabling the identification of more effective treatments. This study, therefore, aims to identify n...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-07-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/14/15/3726 |
| _version_ | 1797442562112356352 |
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| author | Niamh H. McCabe Leanne Stevenson Enya Scanlon Rosalie Douglas Susanna Kennedy Oliver Keminer Björn Windshügel Daniela Zisterer Richard D. Kennedy Jaine K. Blayney Richard C. Turkington |
| author_facet | Niamh H. McCabe Leanne Stevenson Enya Scanlon Rosalie Douglas Susanna Kennedy Oliver Keminer Björn Windshügel Daniela Zisterer Richard D. Kennedy Jaine K. Blayney Richard C. Turkington |
| author_sort | Niamh H. McCabe |
| collection | DOAJ |
| description | Drug resistance limits the effectiveness of oesophageal adenocarcinoma (OAC) chemotherapies, leading to a poor prognosis for this disease. Elucidation of the underlying resistance mechanisms is key to enabling the identification of more effective treatments. This study, therefore, aims to identify novel therapeutic and/or chemotherapy sensitising drug targets in OAC. Transcriptional data from a cohort of 273 pre-treatment OAC biopsies, from patients who received neoadjuvant chemotherapy followed by surgical resection, were analysed using gene set enrichment analysis (GSEA) to determine differential gene expression between responding and non-responding OAC tumours. From this, 80 genes were selected for high-throughput siRNA screening in OAC cell lines with or without standard chemotherapy treatment. In parallel, cell viability assays were performed using a panel of FDA-approved drugs and combination index (CI) values were calculated to evaluate drug synergy with standard chemotherapy. Mechanisms of synergy were investigated using western blot, propidium iodide flow cytometry, and proliferation assays. Taken together, the screens identified that targeting Src, using either siRNA or the small molecule inhibitor dasatinib, enhanced the efficacy of chemotherapy in OAC cells. Further in vitro functional analysis confirmed Src inhibition to be synergistic with standard OAC chemotherapies, 5-fluorouracil (5-FU), and cisplatin (CDDP). In conclusion, a compound screen together with a functional genomic approach identified Src as a potential chemosensitising target in OAC, which could be assessed in a clinical study for poor prognosis OAC patients. |
| first_indexed | 2024-03-09T12:43:40Z |
| format | Article |
| id | doaj.art-4e02f1d7736c4cf7a2810b4cd68bc148 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-09T12:43:40Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-4e02f1d7736c4cf7a2810b4cd68bc1482023-11-30T22:14:26ZengMDPI AGCancers2072-66942022-07-011415372610.3390/cancers14153726Identification of Src as a Therapeutic Target in Oesophageal Adenocarcinoma through Functional Genomic and High-Throughput Drug Screening ApproachesNiamh H. McCabe0Leanne Stevenson1Enya Scanlon2Rosalie Douglas3Susanna Kennedy4Oliver Keminer5Björn Windshügel6Daniela Zisterer7Richard D. Kennedy8Jaine K. Blayney9Richard C. Turkington10Patrick G Johnston Centre for Cancer Research, Queen’s University Belfast, Belfast BT9 7AE, UKPatrick G Johnston Centre for Cancer Research, Queen’s University Belfast, Belfast BT9 7AE, UKPatrick G Johnston Centre for Cancer Research, Queen’s University Belfast, Belfast BT9 7AE, UKPatrick G Johnston Centre for Cancer Research, Queen’s University Belfast, Belfast BT9 7AE, UKPatrick G Johnston Centre for Cancer Research, Queen’s University Belfast, Belfast BT9 7AE, UKFraunhofer Institute for Translational Medicine and Pharmacology ITMP, Discovery Research ScreeningPort, 22525 Hamburg, GermanyFraunhofer Institute for Translational Medicine and Pharmacology ITMP, Discovery Research ScreeningPort, 22525 Hamburg, GermanyTrinity Biomedical Sciences Institute, School of Biochemistry and Immunology, Trinity College Dublin, D08 XW7X Dublin, IrelandAlmac Diagnostics Ltd., Craigavon BT63 5QD, UKPatrick G Johnston Centre for Cancer Research, Queen’s University Belfast, Belfast BT9 7AE, UKPatrick G Johnston Centre for Cancer Research, Queen’s University Belfast, Belfast BT9 7AE, UKDrug resistance limits the effectiveness of oesophageal adenocarcinoma (OAC) chemotherapies, leading to a poor prognosis for this disease. Elucidation of the underlying resistance mechanisms is key to enabling the identification of more effective treatments. This study, therefore, aims to identify novel therapeutic and/or chemotherapy sensitising drug targets in OAC. Transcriptional data from a cohort of 273 pre-treatment OAC biopsies, from patients who received neoadjuvant chemotherapy followed by surgical resection, were analysed using gene set enrichment analysis (GSEA) to determine differential gene expression between responding and non-responding OAC tumours. From this, 80 genes were selected for high-throughput siRNA screening in OAC cell lines with or without standard chemotherapy treatment. In parallel, cell viability assays were performed using a panel of FDA-approved drugs and combination index (CI) values were calculated to evaluate drug synergy with standard chemotherapy. Mechanisms of synergy were investigated using western blot, propidium iodide flow cytometry, and proliferation assays. Taken together, the screens identified that targeting Src, using either siRNA or the small molecule inhibitor dasatinib, enhanced the efficacy of chemotherapy in OAC cells. Further in vitro functional analysis confirmed Src inhibition to be synergistic with standard OAC chemotherapies, 5-fluorouracil (5-FU), and cisplatin (CDDP). In conclusion, a compound screen together with a functional genomic approach identified Src as a potential chemosensitising target in OAC, which could be assessed in a clinical study for poor prognosis OAC patients.https://www.mdpi.com/2072-6694/14/15/3726oesophageal cancerchemotherapydrug resistancesiRNA screen |
| spellingShingle | Niamh H. McCabe Leanne Stevenson Enya Scanlon Rosalie Douglas Susanna Kennedy Oliver Keminer Björn Windshügel Daniela Zisterer Richard D. Kennedy Jaine K. Blayney Richard C. Turkington Identification of Src as a Therapeutic Target in Oesophageal Adenocarcinoma through Functional Genomic and High-Throughput Drug Screening Approaches Cancers oesophageal cancer chemotherapy drug resistance siRNA screen |
| title | Identification of Src as a Therapeutic Target in Oesophageal Adenocarcinoma through Functional Genomic and High-Throughput Drug Screening Approaches |
| title_full | Identification of Src as a Therapeutic Target in Oesophageal Adenocarcinoma through Functional Genomic and High-Throughput Drug Screening Approaches |
| title_fullStr | Identification of Src as a Therapeutic Target in Oesophageal Adenocarcinoma through Functional Genomic and High-Throughput Drug Screening Approaches |
| title_full_unstemmed | Identification of Src as a Therapeutic Target in Oesophageal Adenocarcinoma through Functional Genomic and High-Throughput Drug Screening Approaches |
| title_short | Identification of Src as a Therapeutic Target in Oesophageal Adenocarcinoma through Functional Genomic and High-Throughput Drug Screening Approaches |
| title_sort | identification of src as a therapeutic target in oesophageal adenocarcinoma through functional genomic and high throughput drug screening approaches |
| topic | oesophageal cancer chemotherapy drug resistance siRNA screen |
| url | https://www.mdpi.com/2072-6694/14/15/3726 |
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