Subtype-specific kinase dependency regulates growth and metastasis of poor-prognosis mesenchymal colorectal cancer
Abstract Background Colorectal cancer (CRC) can be divided into four consensus molecular subtypes (CMS), each with distinct biological features. CMS4 is associated with epithelial-mesenchymal transition and stromal infiltration (Guinney et al., Nat Med 21:1350–6, 2015; Linnekamp et al., Cell Death D...
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| Format: | Article |
| Language: | English |
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BMC
2023-03-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
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| Online Access: | https://doi.org/10.1186/s13046-023-02600-9 |
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| author | Joyce Y. Buikhuisen Patricia M. Gomez Barila Kate Cameron Saskia J. E. Suijkerbuijk Cor Lieftink Simone di Franco Ana Krotenberg Garcia Rebeca Uceda Castro Kristiaan J. Lenos Lisanne E. Nijman Arezo Torang Ciro Longobardi Joan H. de Jong Daniëlle Dekker Giorgio Stassi Louis Vermeulen Roderick L. Beijersbergen Jacco van Rheenen Stephan Huveneers Jan Paul Medema |
| author_facet | Joyce Y. Buikhuisen Patricia M. Gomez Barila Kate Cameron Saskia J. E. Suijkerbuijk Cor Lieftink Simone di Franco Ana Krotenberg Garcia Rebeca Uceda Castro Kristiaan J. Lenos Lisanne E. Nijman Arezo Torang Ciro Longobardi Joan H. de Jong Daniëlle Dekker Giorgio Stassi Louis Vermeulen Roderick L. Beijersbergen Jacco van Rheenen Stephan Huveneers Jan Paul Medema |
| author_sort | Joyce Y. Buikhuisen |
| collection | DOAJ |
| description | Abstract Background Colorectal cancer (CRC) can be divided into four consensus molecular subtypes (CMS), each with distinct biological features. CMS4 is associated with epithelial-mesenchymal transition and stromal infiltration (Guinney et al., Nat Med 21:1350–6, 2015; Linnekamp et al., Cell Death Differ 25:616–33, 2018), whereas clinically it is characterized by lower responses to adjuvant therapy, higher incidence of metastatic spreading and hence dismal prognosis (Buikhuisen et al., Oncogenesis 9:66, 2020). Methods To understand the biology of the mesenchymal subtype and unveil specific vulnerabilities, a large CRISPR-Cas9 drop-out screen was performed on 14 subtyped CRC cell lines to uncover essential kinases in all CMSs. Dependency of CMS4 cells on p21-activated kinase 2 (PAK2) was validated in independent 2D and 3D in vitro cultures and in vivo models assessing primary and metastatic outgrowth in liver and peritoneum. TIRF microscopy was used to uncover actin cytoskeleton dynamics and focal adhesion localization upon PAK2 loss. Subsequent functional assays were performed to determine altered growth and invasion patterns. Results PAK2 was identified as a key kinase uniquely required for growth of the mesenchymal subtype CMS4, both in vitro and in vivo. PAK2 plays an important role in cellular attachment and cytoskeletal rearrangements (Coniglio et al., Mol Cell Biol 28:4162–72, 2008; Grebenova et al., Sci Rep 9:17171, 2019). In agreement, deletion or inhibition of PAK2 impaired actin cytoskeleton dynamics in CMS4 cells and, as a consequence, significantly reduced invasive capacity, while it was dispensable for CMS2 cells. Clinical relevance of these findings was supported by the observation that deletion of PAK2 from CMS4 cells prevented metastatic spreading in vivo. Moreover, growth in a model for peritoneal metastasis was hampered when CMS4 tumor cells were deficient for PAK2. Conclusion Our data reveal a unique dependency of mesenchymal CRC and provide a rationale for PAK2 inhibition to target this aggressive subgroup of colorectal cancer. |
| first_indexed | 2024-04-09T22:33:25Z |
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| id | doaj.art-f5507e6ca1b2488d88586c36a3b6eee2 |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-04-09T22:33:25Z |
| publishDate | 2023-03-01 |
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| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-f5507e6ca1b2488d88586c36a3b6eee22023-03-22T12:41:32ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662023-03-0142111810.1186/s13046-023-02600-9Subtype-specific kinase dependency regulates growth and metastasis of poor-prognosis mesenchymal colorectal cancerJoyce Y. Buikhuisen0Patricia M. Gomez Barila1Kate Cameron2Saskia J. E. Suijkerbuijk3Cor Lieftink4Simone di Franco5Ana Krotenberg Garcia6Rebeca Uceda Castro7Kristiaan J. Lenos8Lisanne E. Nijman9Arezo Torang10Ciro Longobardi11Joan H. de Jong12Daniëlle Dekker13Giorgio Stassi14Louis Vermeulen15Roderick L. Beijersbergen16Jacco van Rheenen17Stephan Huveneers18Jan Paul Medema19Laboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCOncode InstituteOncode InstituteDepartment of Surgical Oncological and Stomatological Sciences (DICHIRONS), University of PalermoOncode InstituteOncode InstituteLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCDepartment of Surgical Oncological and Stomatological Sciences (DICHIRONS), University of PalermoLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCOncode InstituteOncode InstituteDepartment of Medical Biochemistry, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of AmsterdamLaboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, location AMCAbstract Background Colorectal cancer (CRC) can be divided into four consensus molecular subtypes (CMS), each with distinct biological features. CMS4 is associated with epithelial-mesenchymal transition and stromal infiltration (Guinney et al., Nat Med 21:1350–6, 2015; Linnekamp et al., Cell Death Differ 25:616–33, 2018), whereas clinically it is characterized by lower responses to adjuvant therapy, higher incidence of metastatic spreading and hence dismal prognosis (Buikhuisen et al., Oncogenesis 9:66, 2020). Methods To understand the biology of the mesenchymal subtype and unveil specific vulnerabilities, a large CRISPR-Cas9 drop-out screen was performed on 14 subtyped CRC cell lines to uncover essential kinases in all CMSs. Dependency of CMS4 cells on p21-activated kinase 2 (PAK2) was validated in independent 2D and 3D in vitro cultures and in vivo models assessing primary and metastatic outgrowth in liver and peritoneum. TIRF microscopy was used to uncover actin cytoskeleton dynamics and focal adhesion localization upon PAK2 loss. Subsequent functional assays were performed to determine altered growth and invasion patterns. Results PAK2 was identified as a key kinase uniquely required for growth of the mesenchymal subtype CMS4, both in vitro and in vivo. PAK2 plays an important role in cellular attachment and cytoskeletal rearrangements (Coniglio et al., Mol Cell Biol 28:4162–72, 2008; Grebenova et al., Sci Rep 9:17171, 2019). In agreement, deletion or inhibition of PAK2 impaired actin cytoskeleton dynamics in CMS4 cells and, as a consequence, significantly reduced invasive capacity, while it was dispensable for CMS2 cells. Clinical relevance of these findings was supported by the observation that deletion of PAK2 from CMS4 cells prevented metastatic spreading in vivo. Moreover, growth in a model for peritoneal metastasis was hampered when CMS4 tumor cells were deficient for PAK2. Conclusion Our data reveal a unique dependency of mesenchymal CRC and provide a rationale for PAK2 inhibition to target this aggressive subgroup of colorectal cancer.https://doi.org/10.1186/s13046-023-02600-9Colorectal cancerEpithelial-mesenchymal transitionMetastasisCellular attachmentPAK family |
| spellingShingle | Joyce Y. Buikhuisen Patricia M. Gomez Barila Kate Cameron Saskia J. E. Suijkerbuijk Cor Lieftink Simone di Franco Ana Krotenberg Garcia Rebeca Uceda Castro Kristiaan J. Lenos Lisanne E. Nijman Arezo Torang Ciro Longobardi Joan H. de Jong Daniëlle Dekker Giorgio Stassi Louis Vermeulen Roderick L. Beijersbergen Jacco van Rheenen Stephan Huveneers Jan Paul Medema Subtype-specific kinase dependency regulates growth and metastasis of poor-prognosis mesenchymal colorectal cancer Journal of Experimental & Clinical Cancer Research Colorectal cancer Epithelial-mesenchymal transition Metastasis Cellular attachment PAK family |
| title | Subtype-specific kinase dependency regulates growth and metastasis of poor-prognosis mesenchymal colorectal cancer |
| title_full | Subtype-specific kinase dependency regulates growth and metastasis of poor-prognosis mesenchymal colorectal cancer |
| title_fullStr | Subtype-specific kinase dependency regulates growth and metastasis of poor-prognosis mesenchymal colorectal cancer |
| title_full_unstemmed | Subtype-specific kinase dependency regulates growth and metastasis of poor-prognosis mesenchymal colorectal cancer |
| title_short | Subtype-specific kinase dependency regulates growth and metastasis of poor-prognosis mesenchymal colorectal cancer |
| title_sort | subtype specific kinase dependency regulates growth and metastasis of poor prognosis mesenchymal colorectal cancer |
| topic | Colorectal cancer Epithelial-mesenchymal transition Metastasis Cellular attachment PAK family |
| url | https://doi.org/10.1186/s13046-023-02600-9 |
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