Mechanical phenotyping of cancer stemness in colorectal cancer
[eng] In this thesis work, I explored the link between cancer stemness and the mechanical properties of colorectal cancer (CRC) cells. In the last decades, the cancer stem cell (CSC) model underwent an important paradigm shift, from a static and intrinsic concept of CSCs to a more dynamic and plasti...
| Autor: | |
|---|---|
| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2023 |
| País: | España |
| Institución: | Universidad de Barcelona |
| Repositorio: | Dipòsit Digital de la UB |
| OAI Identifier: | oai:diposit.ub.edu:2445/223212 |
| Acceso en línea: | https://hdl.handle.net/2445/223212 http://hdl.handle.net/10803/695225 |
| Access Level: | acceso abierto |
| Palabra clave: | Càncer colorectal Cèl·lules mare Metàstasi Colorectal cancer Stem cells Metastasis |
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España |
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Mechanical phenotyping of cancer stemness in colorectal cancer |
| title |
Mechanical phenotyping of cancer stemness in colorectal cancer |
| spellingShingle |
Mechanical phenotyping of cancer stemness in colorectal cancer Conti, Sefora Càncer colorectal Cèl·lules mare Metàstasi Colorectal cancer Stem cells Metastasis |
| title_short |
Mechanical phenotyping of cancer stemness in colorectal cancer |
| title_full |
Mechanical phenotyping of cancer stemness in colorectal cancer |
| title_fullStr |
Mechanical phenotyping of cancer stemness in colorectal cancer |
| title_full_unstemmed |
Mechanical phenotyping of cancer stemness in colorectal cancer |
| title_sort |
Mechanical phenotyping of cancer stemness in colorectal cancer |
| dc.creator.none.fl_str_mv |
Conti, Sefora |
| author |
Conti, Sefora |
| author_facet |
Conti, Sefora |
| author_role |
author |
| dc.contributor.none.fl_str_mv |
Trepat Guixer, Xavier Labernadie, Anna Universitat de Barcelona. Facultat de Medicina i Ciències de la Salut |
| dc.subject.none.fl_str_mv |
Càncer colorectal Cèl·lules mare Metàstasi Colorectal cancer Stem cells Metastasis |
| topic |
Càncer colorectal Cèl·lules mare Metàstasi Colorectal cancer Stem cells Metastasis |
| description |
[eng] In this thesis work, I explored the link between cancer stemness and the mechanical properties of colorectal cancer (CRC) cells. In the last decades, the cancer stem cell (CSC) model underwent an important paradigm shift, from a static and intrinsic concept of CSCs to a more dynamic and plastic notion, integrating the influence of the microenvironment. Similarly, during the last five years, my experimental approach had to shift, from assuming that cancer stem cells are static and thus won’t change in response to different culture conditions, to the realization that their stemness is largely affected by the microenvironment, and thus I had to establish the experimental conditions that would maintain the desired cellular phenotypes. This effort is not reflected in the present work, but it constitutes its foundation. CRC tumors are composed of heterogeneous cell populations including a pool of cancer stem cells (CSCs) that express LGR5. The link between cancer cell differentiation states and their metastatic potential has been the focus of extensive investigation, with some studies pointing to microenvironmentally defined plasticity as a mechanism indispensable for metastasis formation. In this highly heterogeneous and plastic context, differences in mechanical phenotypes may favor or impair the ability of specific cell populations to progress through the metastatic cascade. However, whether distinct cell populations in CRC tumors display different mechanical properties, and how these properties might contribute to metastasis is unknown. In the present study I performed a broad biophysical characterization of CRC patient derived organoids (PDOs) engineered to fluorescently label cells expressing LGR5. I found that LGR5+ and LGR5- cells display distinct mechanical phenotypes. Compared to LGR5- cells, LGR5+ cells are stiffer, adhere better to the extracellular matrix (ECM), move slower both as single cells and clusters, display higher nuclear YAP, and show a higher survival rate in response to mechanical confinement. These differences are largely explained by the downregulation of the membrane to cortex attachment proteins Ezrin/Radixin/Moesin (ERMs) in the LGR5+ cells. By analyzing scRNA-seq expression patterns from a patient cohort, I show that this downregulation is a robust signature of colorectal tumors. Finally, I also show that LGR5+ cells adhere better to the endothelial surface and form transendothelial gaps with higher efficiency than LGR5- cells. Together, these results show that LGR5- cells display a mechanically dynamic phenotype that favors dissemination from the primary tumor whereas LGR5+ cells display a mechanically stable and resilient phenotype that promotes extravasation and metastatic growth. The observed coupling between mechanical states and cancer cell heterogeneity may be an indispensable adaptive mechanism for metastatic progression. |
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2023 |
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2023 |
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info:eu-repo/semantics/doctoralThesis info:eu-repo/semantics/publishedVersion |
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doctoralThesis |
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publishedVersion |
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https://hdl.handle.net/2445/223212 http://hdl.handle.net/10803/695225 |
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https://hdl.handle.net/2445/223212 http://hdl.handle.net/10803/695225 |
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Inglés |
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Inglés |
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(c) Conti, Sefora, 2023 info:eu-repo/semantics/openAccess |
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(c) Conti, Sefora, 2023 |
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openAccess |
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application/pdf |
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Universitat de Barcelona |
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Universitat de Barcelona |
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Tesis Doctorals - Facultat - Medicina i Ciències de la Salut reponame:Dipòsit Digital de la UB instname:Universidad de Barcelona |
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Universidad de Barcelona |
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Dipòsit Digital de la UB |
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Dipòsit Digital de la UB |
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1869413162492624896 |
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Mechanical phenotyping of cancer stemness in colorectal cancerConti, SeforaCàncer colorectalCèl·lules mareMetàstasiColorectal cancerStem cellsMetastasis[eng] In this thesis work, I explored the link between cancer stemness and the mechanical properties of colorectal cancer (CRC) cells. In the last decades, the cancer stem cell (CSC) model underwent an important paradigm shift, from a static and intrinsic concept of CSCs to a more dynamic and plastic notion, integrating the influence of the microenvironment. Similarly, during the last five years, my experimental approach had to shift, from assuming that cancer stem cells are static and thus won’t change in response to different culture conditions, to the realization that their stemness is largely affected by the microenvironment, and thus I had to establish the experimental conditions that would maintain the desired cellular phenotypes. This effort is not reflected in the present work, but it constitutes its foundation. CRC tumors are composed of heterogeneous cell populations including a pool of cancer stem cells (CSCs) that express LGR5. The link between cancer cell differentiation states and their metastatic potential has been the focus of extensive investigation, with some studies pointing to microenvironmentally defined plasticity as a mechanism indispensable for metastasis formation. In this highly heterogeneous and plastic context, differences in mechanical phenotypes may favor or impair the ability of specific cell populations to progress through the metastatic cascade. However, whether distinct cell populations in CRC tumors display different mechanical properties, and how these properties might contribute to metastasis is unknown. In the present study I performed a broad biophysical characterization of CRC patient derived organoids (PDOs) engineered to fluorescently label cells expressing LGR5. I found that LGR5+ and LGR5- cells display distinct mechanical phenotypes. Compared to LGR5- cells, LGR5+ cells are stiffer, adhere better to the extracellular matrix (ECM), move slower both as single cells and clusters, display higher nuclear YAP, and show a higher survival rate in response to mechanical confinement. These differences are largely explained by the downregulation of the membrane to cortex attachment proteins Ezrin/Radixin/Moesin (ERMs) in the LGR5+ cells. By analyzing scRNA-seq expression patterns from a patient cohort, I show that this downregulation is a robust signature of colorectal tumors. Finally, I also show that LGR5+ cells adhere better to the endothelial surface and form transendothelial gaps with higher efficiency than LGR5- cells. Together, these results show that LGR5- cells display a mechanically dynamic phenotype that favors dissemination from the primary tumor whereas LGR5+ cells display a mechanically stable and resilient phenotype that promotes extravasation and metastatic growth. The observed coupling between mechanical states and cancer cell heterogeneity may be an indispensable adaptive mechanism for metastatic progression.[spa] En esta tesis, he explorado la relación entre las células madre cancerosas y las propiedades mecánicas de las células del cáncer de colon. En las últimas décadas, el modelo de células madre de cancerosas ha experimentado un importante cambio de paradigma, pasando de un concepto estático e intrínseco de células madre cancerosas a una noción más dinámica y plástica, que integra la influencia del microambiente. Del mismo modo, durante los últimos cinco años, mi enfoque experimental tuvo que cambiar, pasando de suponer que las células madre cancerosas son estáticas y, por tanto, no cambiarán en respuesta a diferentes condiciones de cultivo, a la constatación de que su carácter de células madre se ve afectado en gran medida por el microambiente, por lo que tuve que establecer las condiciones experimentales que mantuvieran los fenotipos celulares deseados. Este esfuerzo no se refleja en el presente trabajo, pero constituye su fundamento. Los tumores de cáncer colorrectal están compuestos por poblaciones celulares heterogéneas que incluyen un conjunto de células madre cancerosas, marcadas por la expresión de LGR5. La relación entre los estados de diferenciación de las células cancerosas y su potencial metastásico ha sido objeto de una amplia investigación, y algunos estudios apuntan a la plasticidad definida por el microambiente como mecanismo indispensable para la formación de metástasis. En este contexto altamente heterogéneo y plástico, las diferencias en los fenotipos mecánicos pueden favorecer o perjudicar el potencial metastático de las células cancerosas. Sin embargo, se desconoce si las distintas poblaciones celulares de los tumores de colon presentan diferentes propiedades mecánicas y cómo estas propiedades podrían contribuir a las metástasis. En este estudio llevé a cabo una amplia caracterización biofísica de organoides derivados de pacientes de cáncer de colon diseñados para etiquetar con fluorescencia células que expresan LGR5. Descubrí que las células LGR5+ y LGR5- muestran fenotipos mecánicos distintos. En comparación con las células LGR5-, las células LGR5+ son más rígidas, se adhieren mejor a la matriz extracelular, se mueven más lentamente tanto como células individualmente que en grupo, muestran mas YAP nuclear y una mayor tasa de supervivencia en respuesta al confinamiento mecánico. Estas diferencias se explican en gran medida por la regulación a la baja de las proteínas de unión de la membrana a la corteza Ezrin/Radixina/Moesina en las células LGR5+. Mediante el análisis de los patrones de expresión scRNA-seq de una cohorte de pacientes, demuestro que esta regulación a la baja es una firma robusta de los tumores colorrectales. Por último, también demuestro que las células LGR5+ se adhieren mejor a la superficie endotelial y forman brechas transendoteliales con mayor eficacia que las células LGR5-. En conjunto, estos resultados muestran que las células LGR5- presentan un fenotipo mecánicamente dinámico que favorece la diseminación desde el tumor primario, mientras que las células LGR5+ presentan un fenotipo mecánicamente estable y resistente que favorece la extravasación y el crecimiento metastásico. El acoplamiento observado entre los estados mecánicos y la heterogeneidad de las células cancerosas puede ser un mecanismo adaptativo indispensable para la progresión metastásica.Universitat de BarcelonaTrepat Guixer, XavierLabernadie, AnnaUniversitat de Barcelona. Facultat de Medicina i Ciències de la Salut2023info:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/223212http://hdl.handle.net/10803/695225Tesis Doctorals - Facultat - Medicina i Ciències de la Salutreponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglés(c) Conti, Sefora, 2023info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/2232122026-05-27T06:46:51Z |
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15,811543 |