Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical models
Abstract Background Autotaxin (ATX) is a secreted enzyme that converts lysophosphatidylcholine to lysophosphatidic acid (LPA). LPA stimulates cell proliferation and migration and promotes wound repair following tissue damage. ATX levels are directly correlated with stage and grade in several human c...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-08-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13046-023-02780-4 |
| _version_ | 1797555695698051072 |
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| author | Matteo Centonze Giusy Di Conza Michael Lahn Isabel Fabregat Francesco Dituri Isabella Gigante Grazia Serino Rosanna Scialpi Livianna Carrieri Roberto Negro Elena Pizzuto Gianluigi Giannelli |
| author_facet | Matteo Centonze Giusy Di Conza Michael Lahn Isabel Fabregat Francesco Dituri Isabella Gigante Grazia Serino Rosanna Scialpi Livianna Carrieri Roberto Negro Elena Pizzuto Gianluigi Giannelli |
| author_sort | Matteo Centonze |
| collection | DOAJ |
| description | Abstract Background Autotaxin (ATX) is a secreted enzyme that converts lysophosphatidylcholine to lysophosphatidic acid (LPA). LPA stimulates cell proliferation and migration and promotes wound repair following tissue damage. ATX levels are directly correlated with stage and grade in several human cancers. Several small molecule ATX inhibitors have been developed in recent years. IOA-289 is a potent ATX inhibitor, developed to treat cancers containing fibrosis. In this study, we tested IOA-289 treatment on different gastrointestinal tract tumor cell lines, in order to evaluate its effects on viability and motility. Methods To determine the effects on cell viability and proliferation of treatment with increasing concentrations of IOA-289, we used the crystal violet assay, a clonogenic assay in matrigel, and we evaluated the inhibitor’s effect on formation of 3D spheroids in an in vitro model. The effect of IOA-289 on cell cycle phases was analysed with a redox dye reagent. Cell migration capacity was evaluated by wound healing assay and transwell migration assay. To evaluate the pro-apoptotic effect of the inhibitor, cells were stained with Annexin V and immunofluorescence and flow cytometry analysis were performed. An antibody array was also used, to discriminate, in various samples, the differential expression of 43 proteins involved in the apoptosis pathway. Results We found that IOA-289 is able to inhibit both growth and migration of gastrointestinal tract tumor cell lines, both in 2D (crystal violet assay) and 3D in vitro models (spheroid formation and clonogenic assay in matrigel). This effect is dose-dependent, and the drug is most effective when administered in FBS-free culture medium. The inhibitory effect on cell growth is due to a pro-apoptotic effect of IOA-289. Staining with FITC-conjugated Annexin V showed that IOA-289 induced a dose-dependent increase in fluorescence following incubation for 24 h, and apoptotic cells were also distinguished in flow cytometry using Annexin/PI staining. The antibody array shows that treatment with IOA-289 causes the increased expression of several pro-apoptotic proteins in all tested cell lines. Conclusions These results indicate that IOA-289 may be an effective drug for the treatment of tumors of the gastrointestinal tract, particularly those characterized by a high degree of fibrosis. |
| first_indexed | 2024-03-10T16:51:08Z |
| format | Article |
| id | doaj.art-bc5b94d5541c4e2db10ab31f71fec14a |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-03-10T16:51:08Z |
| publishDate | 2023-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-bc5b94d5541c4e2db10ab31f71fec14a2023-11-20T11:18:05ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662023-08-0142111610.1186/s13046-023-02780-4Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical modelsMatteo Centonze0Giusy Di Conza1Michael Lahn2Isabel Fabregat3Francesco Dituri4Isabella Gigante5Grazia Serino6Rosanna Scialpi7Livianna Carrieri8Roberto Negro9Elena Pizzuto10Gianluigi Giannelli11National Institute of Gastroenterology - IRCCS “Saverio de Bellis”iOnctura SAiOnctura SATGF-β and Cancer Group, Oncobell Program, Bellvitge Biomedical Research Institute (IDIBELL) and CIBEREHD – ISCIIINational Institute of Gastroenterology - IRCCS “Saverio de Bellis”National Institute of Gastroenterology - IRCCS “Saverio de Bellis”National Institute of Gastroenterology - IRCCS “Saverio de Bellis”National Institute of Gastroenterology - IRCCS “Saverio de Bellis”National Institute of Gastroenterology - IRCCS “Saverio de Bellis”National Institute of Gastroenterology - IRCCS “Saverio de Bellis”National Institute of Gastroenterology - IRCCS “Saverio de Bellis”National Institute of Gastroenterology - IRCCS “Saverio de Bellis”Abstract Background Autotaxin (ATX) is a secreted enzyme that converts lysophosphatidylcholine to lysophosphatidic acid (LPA). LPA stimulates cell proliferation and migration and promotes wound repair following tissue damage. ATX levels are directly correlated with stage and grade in several human cancers. Several small molecule ATX inhibitors have been developed in recent years. IOA-289 is a potent ATX inhibitor, developed to treat cancers containing fibrosis. In this study, we tested IOA-289 treatment on different gastrointestinal tract tumor cell lines, in order to evaluate its effects on viability and motility. Methods To determine the effects on cell viability and proliferation of treatment with increasing concentrations of IOA-289, we used the crystal violet assay, a clonogenic assay in matrigel, and we evaluated the inhibitor’s effect on formation of 3D spheroids in an in vitro model. The effect of IOA-289 on cell cycle phases was analysed with a redox dye reagent. Cell migration capacity was evaluated by wound healing assay and transwell migration assay. To evaluate the pro-apoptotic effect of the inhibitor, cells were stained with Annexin V and immunofluorescence and flow cytometry analysis were performed. An antibody array was also used, to discriminate, in various samples, the differential expression of 43 proteins involved in the apoptosis pathway. Results We found that IOA-289 is able to inhibit both growth and migration of gastrointestinal tract tumor cell lines, both in 2D (crystal violet assay) and 3D in vitro models (spheroid formation and clonogenic assay in matrigel). This effect is dose-dependent, and the drug is most effective when administered in FBS-free culture medium. The inhibitory effect on cell growth is due to a pro-apoptotic effect of IOA-289. Staining with FITC-conjugated Annexin V showed that IOA-289 induced a dose-dependent increase in fluorescence following incubation for 24 h, and apoptotic cells were also distinguished in flow cytometry using Annexin/PI staining. The antibody array shows that treatment with IOA-289 causes the increased expression of several pro-apoptotic proteins in all tested cell lines. Conclusions These results indicate that IOA-289 may be an effective drug for the treatment of tumors of the gastrointestinal tract, particularly those characterized by a high degree of fibrosis.https://doi.org/10.1186/s13046-023-02780-4AutotaxinLysophosphatidic acidGastrointestinal cancers |
| spellingShingle | Matteo Centonze Giusy Di Conza Michael Lahn Isabel Fabregat Francesco Dituri Isabella Gigante Grazia Serino Rosanna Scialpi Livianna Carrieri Roberto Negro Elena Pizzuto Gianluigi Giannelli Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical models Journal of Experimental & Clinical Cancer Research Autotaxin Lysophosphatidic acid Gastrointestinal cancers |
| title | Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical models |
| title_full | Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical models |
| title_fullStr | Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical models |
| title_full_unstemmed | Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical models |
| title_short | Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical models |
| title_sort | autotaxin inhibitor ioa 289 reduces gastrointestinal cancer progression in preclinical models |
| topic | Autotaxin Lysophosphatidic acid Gastrointestinal cancers |
| url | https://doi.org/10.1186/s13046-023-02780-4 |
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