FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis
Abstract Background Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that is overexpressed or activated in several advanced-stage solid cancers. It is known to play both kinase-dependent and -independent roles in promoting tumor progression and metastasis. Numerous inhibitors, targeting...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-01-01
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| Series: | Cell Communication and Signaling |
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| Online Access: | https://doi.org/10.1186/s12964-020-00671-1 |
| _version_ | 1818951221354954752 |
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| author | Ioanna Antoniades Maria Kyriakou Anna Charalambous Katerina Kalalidou Andri Christodoulou Maria Christoforou Paris A. Skourides |
| author_facet | Ioanna Antoniades Maria Kyriakou Anna Charalambous Katerina Kalalidou Andri Christodoulou Maria Christoforou Paris A. Skourides |
| author_sort | Ioanna Antoniades |
| collection | DOAJ |
| description | Abstract Background Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that is overexpressed or activated in several advanced-stage solid cancers. It is known to play both kinase-dependent and -independent roles in promoting tumor progression and metastasis. Numerous inhibitors, targeting either the enzymatic or scaffolding activities of FAK have been generated, with varying degree of success. Here, we describe a novel approach to site-specifically target both kinase-dependent and -independent FAK functions at focal adhesions (FAs), the primary sites at which the kinase exerts its activity. Methods We took advantage of the well-characterized interactions between the paxillin LD motifs and the FAK FAT domain and generated a polypeptide (LD2-LD3-LD4) expected to compete with interactions with paxillin. Co-immunoprecipitation experiments were performed to examine the interaction between the LD2-LD3-LD4 polypeptide and FAK. The effects of LD2-LD3-LD4 in the localization and functions of FAK, as well as FA composition, were evaluated using quantitative immunofluorescence, cell fractionation, FA isolation and Western Blot analysis. Live cell imaging, as well as 2-D migration and cell invasion assays were used to examine the effects on FA turnover and tumor cell migration and invasion. Results Expression of the LD2-LD3-LD4 polypeptide prevents FAK localization at FAs, in a controlled and dose-dependent manner, by competing with endogenous paxillin for FAK binding. Importantly, the LD2-LD3-LD4 peptide did not otherwise affect FA composition or integrin activation. LD2-LD3-LD4 inhibited FAK-dependent downstream integrin signaling and, unlike existing inhibitors, also blocked FAK’s scaffolding functions. We further show that LD2-LD3-LD4 expression markedly reduces FA turnover and inhibits tumor cell migration and invasion. Finally, we show that dimers of a single motif, linked through a flexible linker of the proper size, are sufficient for the displacement of FAK from FAs and for inhibition of tumor cell migration. This work raises the possibility of using a synthetic peptide as an antimetastatic agent, given that effective displacement of FAK from FAs only requires dimers of a single LD motif linked by a short flexible linker. Conclusion In conclusion, these results suggest that FAK displacement from FAs is a promising new strategy to target critical processes implicated in cancer progression and metastasis. Video abstract. Graphical abstract |
| first_indexed | 2024-12-20T09:31:02Z |
| format | Article |
| id | doaj.art-50f8f3c833674dccaaf12b8c6d633226 |
| institution | Directory Open Access Journal |
| issn | 1478-811X |
| language | English |
| last_indexed | 2024-12-20T09:31:02Z |
| publishDate | 2021-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj.art-50f8f3c833674dccaaf12b8c6d6332262022-12-21T19:45:05ZengBMCCell Communication and Signaling1478-811X2021-01-0119112210.1186/s12964-020-00671-1FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasisIoanna Antoniades0Maria Kyriakou1Anna Charalambous2Katerina Kalalidou3Andri Christodoulou4Maria Christoforou5Paris A. Skourides6Department of Biological Sciences, University of CyprusDepartment of Biological Sciences, University of CyprusDepartment of Biological Sciences, University of CyprusDepartment of Biological Sciences, University of CyprusDepartment of Biological Sciences, University of CyprusDepartment of Biological Sciences, University of CyprusDepartment of Biological Sciences, University of CyprusAbstract Background Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that is overexpressed or activated in several advanced-stage solid cancers. It is known to play both kinase-dependent and -independent roles in promoting tumor progression and metastasis. Numerous inhibitors, targeting either the enzymatic or scaffolding activities of FAK have been generated, with varying degree of success. Here, we describe a novel approach to site-specifically target both kinase-dependent and -independent FAK functions at focal adhesions (FAs), the primary sites at which the kinase exerts its activity. Methods We took advantage of the well-characterized interactions between the paxillin LD motifs and the FAK FAT domain and generated a polypeptide (LD2-LD3-LD4) expected to compete with interactions with paxillin. Co-immunoprecipitation experiments were performed to examine the interaction between the LD2-LD3-LD4 polypeptide and FAK. The effects of LD2-LD3-LD4 in the localization and functions of FAK, as well as FA composition, were evaluated using quantitative immunofluorescence, cell fractionation, FA isolation and Western Blot analysis. Live cell imaging, as well as 2-D migration and cell invasion assays were used to examine the effects on FA turnover and tumor cell migration and invasion. Results Expression of the LD2-LD3-LD4 polypeptide prevents FAK localization at FAs, in a controlled and dose-dependent manner, by competing with endogenous paxillin for FAK binding. Importantly, the LD2-LD3-LD4 peptide did not otherwise affect FA composition or integrin activation. LD2-LD3-LD4 inhibited FAK-dependent downstream integrin signaling and, unlike existing inhibitors, also blocked FAK’s scaffolding functions. We further show that LD2-LD3-LD4 expression markedly reduces FA turnover and inhibits tumor cell migration and invasion. Finally, we show that dimers of a single motif, linked through a flexible linker of the proper size, are sufficient for the displacement of FAK from FAs and for inhibition of tumor cell migration. This work raises the possibility of using a synthetic peptide as an antimetastatic agent, given that effective displacement of FAK from FAs only requires dimers of a single LD motif linked by a short flexible linker. Conclusion In conclusion, these results suggest that FAK displacement from FAs is a promising new strategy to target critical processes implicated in cancer progression and metastasis. Video abstract. Graphical abstracthttps://doi.org/10.1186/s12964-020-00671-1Focal adhesion kinase (FAK)PaxillinFocal adhesionsCell migrationCell invasionCancer |
| spellingShingle | Ioanna Antoniades Maria Kyriakou Anna Charalambous Katerina Kalalidou Andri Christodoulou Maria Christoforou Paris A. Skourides FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis Cell Communication and Signaling Focal adhesion kinase (FAK) Paxillin Focal adhesions Cell migration Cell invasion Cancer |
| title | FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis |
| title_full | FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis |
| title_fullStr | FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis |
| title_full_unstemmed | FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis |
| title_short | FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis |
| title_sort | fak displacement from focal adhesions a promising strategy to target processes implicated in cancer progression and metastasis |
| topic | Focal adhesion kinase (FAK) Paxillin Focal adhesions Cell migration Cell invasion Cancer |
| url | https://doi.org/10.1186/s12964-020-00671-1 |
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