Search

The transcription factor Twist1 is involved in the epithelial–mesenchymal transition and contributes to cancer metastasis through mostly unknown mechanisms. In colorectal cancer, Twist1 expression is mainly restricted to the tumor stroma. We found that human fibroblast cell lines stably transfected with Twist1 acquired characteristics of activated cancer-associated fibroblasts (CAFs), such as hyperproliferation, an increased ability to migrate and an alignment of the actin cytoskeleton. Further, Twist1-activated fibroblasts promoted increased matrix stiffness. Using quantitative proteomics, we identified palladin and collagen α1(VI) as two major mediators of the Twist1 effects in fibroblast cell lines. Co-immunoprecipitation studies indicated that palladin and Twist1 interact within the nucleus, suggesting that palladin could act as a transcription regulator. Palladin was found to be more relevant for the cellular biomechanical properties, orientation and polarity, and collagen α1(VI) for the migration and invasion capacity, of Twist1-activated fibroblasts. Both palladin and collagen α1(VI) were observed to be overexpressed in colorectal CAFs and to be associated with poor colorectal cancer patient survival and relapse prediction. Our results demonstrate that Twist1-expressing fibroblasts mimic the properties of CAFs present at the tumor invasive front, which likely explains the prometastatic activities of Twist1. Twist1 appears to require both palladin and collagen α1(VI) as downstream effectors for its prometastatic effects, which could be future therapeutic targets in cancer metastasis., IG-P was supported by a contract S2010/BMD-2344/ Colomics2 from the Comunidad de Madrid. ST was a recipient of a Juan de la Cierva programme. RAB was supported by a grant to established research groups of the Asociación Española Contra el Cáncer (AECC). ML-L was a recipient of a ProteoRed contract. AP-G and BE-P were FPI fellows from the Ministry of Economy and Competitiveness (MINECO). This research was supported by grants to established research groups of the S2010/BMD-2344/Colomics2 from the Comunidad de Madrid, ‘Asociación Española Contra el Cancer (AECC)’, BIO2012-31023 from the MINECO, PRB2 (IPT13/0001-ISCIII-SGEFI/FEDER), RD12/0036/0041 and RD12/0036/0021 from the Instituto de Salud Carlos III-FEDER.

37 p.-6 fig., Pancreatic adenocarcinoma–upregulated factor (PAUF), also known as ZG16B, was previously found in the secretome of metastatic colorectal cancer cells. Here, we demonstrated the presence of PAUF at the intracellular level and its multiple effects on cancer progression. An initial decline of PAUF expression was observed at early stages of colorectal cancer followed by an increase at the metastastic site. PAUF was located at different cellular compartments: membrane-associated vesicles, endosomes, microtubule-associated vesicles, cell growth cones and the cell nucleus. PAUF loss in two colorectal cancer cell lines caused severe alterations in the cell phenotype and cell cycle, including tetraploidy, extensive genomic alterations, micronuclei, and increased apoptosis. An exhaustive analysis of the PAUF interactome using different proteomic approaches revealed the presence of multiple components of the cell cycle, mitotic checkpoint, Wnt pathway and intracellular transport. Among the interacting proteins we found ZW10, a moonlighting protein with a dual function in membrane trafficking and mitosis. In addition, PAUF silencing was associated to APC loss and increased β-catenin nuclear expression. Altogether, our results suggest that PAUF depletion increases aneuploidy, promotes apoptosis and activates the Wnt/β-catenin pathway in colorectal cancer cells facilitating cancer progression. In summary, PAUF behaves as a multifunctional protein, with different roles in cancer progression according to the extra- or intracellular expression, suggesting a therapeutic value for colorectal cancer., This research was supported by grants from the MINECO (BIO2015-66489-R), Fundación Areces and PRB3 (IPT17/0019 - ISCIII-SGEFI /ERDF).

Introduction :Colorectal cancer (CRC) is the second most common cancer in the Western world. About one million of new CRC cases are diagnosed per year. The survival rate of patients with CRC changes widely, even among patients with the same tumor histology. This is possibly due to the impact of environmental factors on tumor development. The diet is the most important among these factors. It is known that nutrition can modify the risk of CRC. However, the role that nutrition plays in the risk of recurrence or survival in patients with CRC is not known with accuracy. Objetive: The objective of this review was to try to clarify this fact. Material and methods: Collecting the data obtained as of today in the epidemiological studies of association among recurrence, survival or mortality risk of CRC; and vitamins and the body mass index (BMI), to develop a series of nutritional advices to patients. Conclusions: Thanks to the studies discussed in this work, we could conclude that: BMI, retinol and vitamin D, and in some cases folic acid, at the time of disease diagnosis, operate like recurrence and survival prognosis markers in CRC.Introducción: el cáncer colorrectal (CCR) es el segundo cáncer más frecuente en el mundo occidental, de tal manera que se diagnostican cerca de 1 millón de casos nuevos por año. La tasa de supervivencia de pacientes con CCR varía ampliamente, aun entre pacientes con el mismo tumor histológico. Esto posiblemente es debido al impacto de los factores ambientales sobre el desarrollo tumoral. Dentro de estos factores, destaca la dieta. Se conoce que la nutrición puede modificar el riesgo de padecer CCR. Sin embargo, no se conoce con precisión el papel que desempeña la nutrición en el riesgo de recurrencia o supervivencia en pacientes con CCR. Objetivo: el objetivo de esta revisión fue tratar de esclarecer el papel que desempeña la nutrición en el riesgo de recurrencia o supervivencia en pacientes con CCR. Material y métodos: recopilación de los datos obtenidos hasta el momento en los estudios epidemiológicos más recientes, de la asociación entre recurrencia, supervivencia o riesgo de mortalidad al CCR; y las vitaminas y el índice de masa corporal (IMC), con el fin de elaborar una serie de consejos nutricionales a estos pacientes. Conclusiones: gracias a los estudios comentados se puede concluir que el IMC y el nivel de vitamina D, retinol y en algunos casos el del ácido fólico, en el momento del diagnóstico de la enfermedad, funcionan como marcadores de pronóstico de recurrencia y supervivencia al CCR.

Colorectal cancer (CRC) is the second most common cancer in the Western world. About one million of new CRC cases are diagnosed per year. The survival rate of patients with CRC changes widely, even among patients with the same tumor histology. This is possibly due to the impact of environmental factors on tumor development. The diet is the most important among these factors. It is known that nutrition can modify the risk of CRC. However, the role that nutrition plays in the risk of recurrence or survival in patients with CRC is not known with accuracy. The objective of this review was to try to clarify this fact. Material and Methods: Collecting the data obtained as of today in the epidemiological studies of association among recurrence, survival or mortality risk of CRC; and vitamins and the body mass index (BMI), to develop a series of nutritional advices to patients. Thanks to the studies discussed in this work, we could conclude that: BMI, retinol and vitamin D, and in some cases folic acid, at the time of disease diagnosis, operate like recurrence and survival prognosis markers in CRC. El cáncer colorrectal (CCR) es el segundo cáncer más frecuente en el mundo occidental, de tal manera que se diagnostican cerca de un millón de casos nuevos por año. La tasa de supervivencia de pacientes con CCR varía ampliamente, aún entre pacientes con el mismo tumor histológico. Esto posiblemente es debido al impacto de los factores ambientales sobre el desarrollo tumoral. Dentro de estos factores, destaca la dieta. Se conoce que la nutrición puede modificar el riesgo de padecer CCR. Sin embargo, no se conoce con precisión el papel que desempeña la nutrición en el riesgo de recurrencia o supervivencia en pacientes con CCR. El objetivo de esta revisión fue tratar de esclarecer el papel que desempeña la nutrición en el riesgo de recurrencia o supervivencia en pacientes con CCR. Material y métodos: recopilación de los datos obtenidos hasta el momento en los estudios epidemiológicos más recientes, de la asociación entre recurrencia, supervivencia o riesgo de mortalidad en el CCR; y las vitaminas y el índice de masa corporal (IMC), con el fin de elaborar una serie de consejos nutricionales a estos pacientes. Gracias a los estudios comentados se puede concluir que el IMC y el nivel de vitamina D, retinol y en algunos casos el del ácido fólico, en el momento del diagnóstico de la enfermedad, funcionan como marcadores de pronóstico de recurrencia y supervivencia al CCR. El càncer colorectal (CCR) és el segon càncer més freqüent en el món occidental, de tal manera que es diagnostiquen prop d'un milió de casos nous per any. La taxa de supervivència de pacients amb CCR varia àmpliament, fins i tot entre pacients amb el mateix tumor histològic. Això possiblement és degut a l'impacte dels factors ambientals sobre el desenvolupament tumoral. Dins d'aquests factors destaca la dieta. Es coneix que la nutrició pot modificar el risc de patir CCR. No obstant això, no es coneix amb precisió el paper que exerceix la nutrició en el risc de recurrència o supervivència en pacients amb CCR. L'objectiu d'aquesta revisió va ser intentar aclarir el paper que exerceix la nutrició en el risc de recurrència o supervivència en pacients amb CCR. Material i mètodes: recopilació de les dades obtingudes fins al moment en els estudis epidemiològics més recents, de l'associació entre recurrència, supervivència o risc de mortalitat en el CCR; i les vitamines i l'índex de massa corporal (IMC), per tal d'elaborar una sèrie de consells nutricionals a aquests pacients. Gràcies als estudis comentats es pot concloure que l'IMC i el nivell de vitamina D, retinol i en alguns casos el de l'àcid fòlic, en el moment del diagnòstic de la malaltia, funcionen com a marcadors de pronòstic de recurrència i supervivència al CCR.

13 p.-6 fig., We have investigated the role of vascular-endothelial (VE)-cadherin in melanoma and breast cancer metastasis. We found that VE-cadherin is expressed in highly aggressive melanoma and breast cancer cell lines. Remarkably, inactivation of VEcadherin triggered a significant loss of malignant traits (proliferation, adhesion, invasion and transendothelial migration) in melanoma and breast cancer cells. These effects, except transendothelial migration, were induced by the VE-cadherin RGD motifs. Co-immunoprecipitation experiments demonstrated an interaction between VE-cadherin and α2β1 integrin, with the RGD motifs found to directly affect β1 integrin activation. VE-cadherin-mediated integrin signaling occurred through specific activation of SRC, ERK and JNK, including AKT in melanoma. Knocking down VEcadherin suppressed lung colonization capacity of melanoma or breast cancer cells inoculated in mice, while pre-incubation with VE-cadherin RGD peptides promoted lung metastasis for both cancer types. Finally, an in silico study revealed the association of high VE-cadherin expression with poor survival in a subset of melanoma patients and breast cancer patients showing low CD34 expression. These findings support a general role for VE-cadherin and other RGD cadherins as critical regulators of lung and liver metastasis in multiple solid tumours. These results pave the way for cadherin-specific RGD targeted therapies to control disseminated metastasis in multiple cancers., BEP was an FPI fellow from Ministry of Economy and Competitiveness (MINECO). This research was supported by grants BIO2012-31023 and BIO2015-66849 from MINECO and PRB2 (IPT13/0001-ISCIII-SGEFI/FEDER) to JIC.

15 p.-9 fig., Little is known about the mechanism of integrin activation by cadherin 17 (CDH17). Here we observed the presence of a tri-peptide motif, RGD, in domain 6 of the human CDH17 sequence and other cadherins such as cadherin 5 and cadherin 6. The use of CDH17 RAD mutants demonstrated a considerable decrease of proliferation and adhesion in RKO and KM12SM colon cancer cells. Furthermore, RGD peptides inhibited the adhesion of both cell lines to recombinant CDH17 domain 6. The RGD motif added exogenously to the cells provoked a change in β1 integrin to an active, high-affinity conformation and an increase in focal adhesion kinase and ERK1/2 activation. In vivo experiments with Swiss nude mice demonstrated that cancer cells expressing the CDH17 RAD mutant showed a considerable delay in tumor growth and liver homing. CDH17 RGD effects were also active in pancreatic cancer cells. Our results suggest that α2β1 integrin interacts with two different ligands, collagen IV and CDH17, using two different binding sites. In summary, the RGD binding motif constitutes a switch for integrin pathway activation and shows a novel capacity of CDH17 as an integrin ligand. This motif could be targeted to avoid metastatic dissemination in tumors overexpressing CDH17 and other RGD-containing cadherins., This research was supported by grant BIO2012-31023 from the Spanish Ministry of Economy and Competitiveness, by a grant to established research groups (AECC), and by grant S2011/BMD-2344/ (Colomics2) from Comunidad de Madrid and ProteoRed-ISCIII.

Most colorectal cancer deaths occur as a consequence of late diagnosis and the appearance of metastasis. We demonstrated that a RGD motif present in liver-intestine cadherin, also known as cadherin 17 (CDH17), plays a critical role in the liver metastasis of colorectal cancer by interacting with α2β1 integrin. CDH17 is able to induce a conformational change in β1 integrin to promote a high affinity conformation. This change is dependent of the RGD motif present in domain 6 of CDH17. The same activation was observed after cell exposition to 9 amino acid peptides containing the RGD motif and the flanking sequences from CDH17. Commercial antibodies anti-CDH17 did not cause any effect on β1 integrin activation. Here, we have used peptides containing the RGD motif and their flanking sequences in CDH17 to retrieve four highly selective monoclonal antibodies with anti-metastatic activity. Only CDH17 RGD-specific monoclonal antibodies inhibited β1 integrin activation in colorectal cancer cells. This inhibition provoked a significant reduction in cell adhesion and proliferation of metastatic cells. In addition, the treatment with these four anti-cadherin RGD antibodies impaired the activation of FAK, JNK and ERK kinases, which are critical for cell adhesion and proliferation. Moreover, RGD-specific monoclonal antibodies were able to cause a significant increase in mouse survival after intra-splenic injection together with highly metastatic cells, as determined by Kaplan-Meier survival analysis and log-rank curves. However, this effect on survival was different for each monoclonal antibody, suggesting a relevant impact of the RGD binding orientation in antibody recognition. As a conclusion, we propose that blocking the interaction between RGD cadherins and α2β1 integrin with highly selective monoclonal antibodies constitutes a promising therapy against colon cancer progression and liver metastasis. Citation Format: Rubén Bartolomé, Sofia Torres, Beatriz Escudero-Paniagua, Carmen Aizpurua, Eva Calviño, Juan I. Imbaud, Ignacio Casal. Anti-metastatic activity of RGD-specific cadherin-17-targeting monoclonal antibodies in colorectal cancer. [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer: From Initiation to Outcomes; 2016 Sep 17-20; Tampa, FL. Philadelphia (PA): AACR; Cancer Res 2017;77(3 Suppl):Abstract nr B15.