Nuevas terapias en cáncer colorrectal metastásico
[EN] Introduction Colorectal cancer (CRC) remains one of the most commonly diagnosed cancers, ranking fourth in terms of incidence and third in terms of mortality. More than 90% of cancer-associated deaths are due to the development of metastasis, a process favored by the acquisition of stem cell-li...
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2022 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/352414 |
| Acceso en línea: | http://hdl.handle.net/10261/352414 |
| Access Level: | acceso abierto |
| Palabra clave: | Cáncer colorrectal Metástasis http://metadata.un.org/sdg/3 Ensure healthy lives and promote well-being for all at all ages |
| Sumario: | [EN] Introduction Colorectal cancer (CRC) remains one of the most commonly diagnosed cancers, ranking fourth in terms of incidence and third in terms of mortality. More than 90% of cancer-associated deaths are due to the development of metastasis, a process favored by the acquisition of stem cell-like properties and cellular plasticity, including epithelial- mesenchymal transition. The metastatic cascade involves different steps involving growth factors, receptors, proteases, chemokines and cell adhesion molecules, such as cadherins and integrins that could be used as therapeutic targets. Currently, there are no effective treatments for the long-term survival of patients with CRC metastasis. This is due to the scarcity of therapeutic targets in metastasis and to the generation of resistance to both chemotherapy and current immunotherapy. For this reason, it is necessary to search for, identify and develop new targets that could be used in the treatment of CRC metastasis. Recently, our group identified relevant proteins in CRC metastasis such as cadherin 17 (CDH17) and interleukin 13 receptor α2 (IL13Rα2). CDH17 contains an RGD motif capable of activating α2β1 integrin, increasing tumor adhesion and proliferation and playing a critical role in tumor growth during liver colonization. Moreover, CDH17 together with the α2β1 integrin appears to participate in the mechanisms of resistance to chemotherapy. On the other hand, IL-13 promotes the synthesis and activation of IL13Rα2 in CRC metastasis. FAM120A and PTP1B are key mediators in the activation of the IL-13/IL13Rα2 pathway and, thus, in FAK and PI3K/AKT/mTOR signaling, leading to increased cell invasion and liver metastasis in colon cancer. Objectives and results Based on the two identified targets, CDH17 and IL13Rα2, in this PhD Thesis we set as main objectives i) to develop new therapeutic tools, such as monoclonal antibodies (MAbs) and peptides, to control metastatic dissemination in different organs and ii) to improve our understanding of the role of CDH17 and IL13Rα2 in CRC metastasis. Among others, the following results were obtained: We obtained a panel of RGD cadherin-specific MAbs selected for their ability to block β1 integrin activation, which inhibited FAK, JNK and ERK signaling, cell adhesion and proliferation in CRC, melanoma, pancreatic and breast cancer. Two of the MAbs, 6.6.1 and 25.4.1, increased the survival of mice to liver and lung metastasis of CRC and melanoma, and delayed lung metastasis of breast cancer cells. Based on these results, MAb 6.6.1 was selected for the humanization process. From 25 humanized variants of MAb 6.6.1, H2L5 was selected, which exhibited the same antimetastatic properties: it inhibited β1 integrin activation, adhesion, cell invasion, migration and proliferation in CRC, melanoma and breast cancer cells. H2L5 also increased mouse survival and reduced liver metastasis of CRC and melanoma cells. To gain insight into the mechanisms of action of anti-RGD cadherin MAbs, quantitative proteomics experiments were performed on cells treated with MAb 6.6.1. Alterations in the proteome of CRC cells treated with MAb 6.6.1 indicated deregulation of proteins involved in the Wnt/catenin pathway, proteosome, actin cytoskeleton or vesicle transport. In addition, we demonstrated that VPS29, a component of the retromer complex, participates in the recycling of CDH17 to membrane. On the other hand, we studied the potential involvement of CDH17 in the mechanisms of irinotecan resistance. Treated and irinotecan-resistant cells showed an increase in CDH17 expression, demonstrating that CDH17 could serve as a predictive biomarker of irinotecan resistance and as a therapeutic target of anti-RGD MAbs in resistant cells. In the case of IL13Rα2, we designed and characterized the D1 peptide corresponding to the IL-13 binding site on IL13Rα2. This peptide contains a highly conserved sequence across mammalian species. The D1 peptide inhibited migration, invasion and proliferation in IL-13-treated metastatic CRC and glioblastoma cells. In addition, it blocked ligand-mediated receptor internalization and degradation, and substantially decreased the signaling ability of IL-13 through IL13Rα2 to activate FAK, PI3K/AKT and Src, as well as that of IL13Rα1 to activate STAT6. Furthermore, D1 treatment induced in vivo protection against CRC cells in liver metastases and arrest glioblastoma xenograft growth. Given the good results obtained with the D1 peptide, it was used to prepare specific MAbs. The anti-IL13Rα2 D1 MAbs were able to inhibit CRC cell adhesion, migration, invasion and proliferation. In addition, they inhibited IL-13-mediated FAK, Src and AKT signaling but not IL13Rα1-mediated STAT6 activation. Although AM 5.5.4 partially blocked IL-13-mediated receptor internalization from the tumor cell surface, it still promoted receptor degradation. MAb 5.5.4 was highly effective in inhibiting in vivo metastatic growth of liver metastases, including established metastases in CRC. Conclusions Both anti-RGD cadherin MAbs and D1 peptide and anti-IL13Rα2 MAbs inhibited the metastatic properties of tumor cells and significantly increased the survival of mice to liver and/or lung metastasis. The mechanism of action of MAb 6.6.1 included proteins involved in the Wnt/-catenin pathway and the VPS29 protein. Furthermore, CDH17 may serve as a biomarker and therapeutic target for irinotecan resistance. In summary, in this PhD Thesis, different therapeutic strategies based on MAbs and peptides have been established against two new targets: the RGD motifs of cadherins (CDH17, VE- cadherin) and the IL13Rα2 receptor, providing sufficient evidences that support their possible clinical application for the treatment of metastatic CRC and, likely, other solid tumors. |
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