Adria Canellas-Socias

All Publications

• Mechanisms of metastatic colorectal cancer. Nature reviews. Gastroenterology & hepatology Cañellas-Socias, A., Sancho, E., Batlle, E. 2024

  Abstract

  Despite extensive research and improvements in understanding colorectal cancer (CRC), its metastatic form continues to pose a substantial challenge, primarily owing to limited therapeutic options and a poor prognosis. This Review addresses the emerging focus on metastatic CRC (mCRC), which has historically been under-studied compared with primary CRC despite its lethality. We delve into two crucial aspects: the molecular and cellular determinants facilitating CRC metastasis and the principles guiding the evolution of metastatic disease. Initially, we examine the genetic alterations integral to CRC metastasis, connecting them to clinically marked characteristics of advanced CRC. Subsequently, we scrutinize the role of cellular heterogeneity and plasticity in metastatic spread and therapy resistance. Finally, we explore how the tumour microenvironment influences metastatic disease, emphasizing the effect of stromal gene programmes and the immune context. The ongoing research in these fields holds immense importance, as its future implications are projected to revolutionize the treatment of patients with mCRC, hopefully offering a promising outlook for their survival.

  View details for DOI 10.1038/s41575-024-00934-z

  View details for PubMedID 38806657

  View details for PubMedCentralID 3563263

• Metastatic recurrence in colorectal cancer arises from residual EMP1(+) cells NATURE Canellas-Socias, A., Cortina, C., Hernando-Momblona, X., Palomo-Ponce, S., Mulholland, E. J., Turon, G., Mateo, L., Conti, S., Roman, O., Sevillano, M., Slebe, F., Stork, D., Caballe-Mestres, A., Berenguer-Llergo, A., Alvarez-Varela, A., Fenderico, N., Novellasdemunt, L., Jimenez-Gracia, L., Sipka, T., Bardia, L., Lorden, P., Colombelli, J., Heyn, H., Trepat, X., Tejpar, S., Sancho, E., Tauriello, D. F., Leedham, S., Attolini, C., Batlle, E. 2022; 611 (7936): 603-+

  Abstract

  Around 30-40% of patients with colorectal cancer (CRC) undergoing curative resection of the primary tumour will develop metastases in the subsequent years1. Therapies to prevent disease relapse remain an unmet medical need. Here we uncover the identity and features of the residual tumour cells responsible for CRC relapse. An analysis of single-cell transcriptomes of samples from patients with CRC revealed that the majority of genes associated with a poor prognosis are expressed by a unique tumour cell population that we named high-relapse cells (HRCs). We established a human-like mouse model of microsatellite-stable CRC that undergoes metastatic relapse after surgical resection of the primary tumour. Residual HRCs occult in mouse livers after primary CRC surgery gave rise to multiple cell types over time, including LGR5+ stem-like tumour cells2-4, and caused overt metastatic disease. Using Emp1 (encoding epithelial membrane protein 1) as a marker gene for HRCs, we tracked and selectively eliminated this cell population. Genetic ablation of EMP1high cells prevented metastatic recurrence and mice remained disease-free after surgery. We also found that HRC-rich micrometastases were infiltrated with T cells, yet became progressively immune-excluded during outgrowth. Treatment with neoadjuvant immunotherapy eliminated residual metastatic cells and prevented mice from relapsing after surgery. Together, our findings reveal the cell-state dynamics of residual disease in CRC and anticipate that therapies targeting HRCs may help to avoid metastatic relapse.

  View details for DOI 10.1038/s41586-022-05402-9

  View details for Web of Science ID 000882576600001

  View details for PubMedID 36352230

  View details for PubMedCentralID 5494503

• TGF beta drives immune evasion in genetically reconstituted colon cancer metastasis NATURE Tauriello, D. F., Palomo-Ponce, S., Stork, D., Berenguer-Llergo, A., Badia-Ramentol, J., Iglesias, M., Sevillano, M., Ibiza, S., Canellas, A., Hernando Momblona, X., Byrom, D., Matarin, J. A., Calon, A., Rivas, E. I., Nebreda, A. R., Riera, A., Stephan-Otto Attolini, C., Batlle, E. 2018; 554 (7693): 538-+

  Abstract

  Most patients with colorectal cancer die as a result of the disease spreading to other organs. However, no prevalent mutations have been associated with metastatic colorectal cancers. Instead, particular features of the tumour microenvironment, such as lack of T-cell infiltration, low type 1 T-helper cell (TH1) activity and reduced immune cytotoxicity or increased TGFβ levels predict adverse outcomes in patients with colorectal cancer. Here we analyse the interplay between genetic alterations and the tumour microenvironment by crossing mice bearing conditional alleles of four main colorectal cancer mutations in intestinal stem cells. Quadruple-mutant mice developed metastatic intestinal tumours that display key hallmarks of human microsatellite-stable colorectal cancers, including low mutational burden, T-cell exclusion and TGFβ-activated stroma. Inhibition of the PD-1-PD-L1 immune checkpoint provoked a limited response in this model system. By contrast, inhibition of TGFβ unleashed a potent and enduring cytotoxic T-cell response against tumour cells that prevented metastasis. In mice with progressive liver metastatic disease, blockade of TGFβ signalling rendered tumours susceptible to anti-PD-1-PD-L1 therapy. Our data show that increased TGFβ in the tumour microenvironment represents a primary mechanism of immune evasion that promotes T-cell exclusion and blocks acquisition of the TH1-effector phenotype. Immunotherapies directed against TGFβ signalling may therefore have broad applications in treating patients with advanced colorectal cancer.

  View details for DOI 10.1038/nature25492

  View details for Web of Science ID 000425597400049

  View details for PubMedID 29443964

• Membrane to cortex attachment determines different mechanical phenotypes in LGR5+ and LGR5- colorectal cancer cells. Nature communications Conti, S., Venturini, V., Canellas-Socias, A., Cortina, C., Abenza, J. F., Stephan-Otto Attolini, C., Middendorp Guerra, E., Xu, C. K., Li, J. H., Rossetti, L., Stassi, G., Roca-Cusachs, P., Diz-Munoz, A., Ruprecht, V., Guck, J., Batlle, E., Labernadie, A., Trepat, X. 2024; 15 (1): 3363

  Abstract

  Colorectal cancer (CRC) tumors are composed of heterogeneous and plastic cell populations, including a pool of cancer stem cells that express LGR5. Whether these distinct cell populations display different mechanical properties, and how these properties might contribute to metastasis is poorly understood. Using CRC patient derived organoids (PDOs), we find that compared to LGR5- cells, LGR5+ cancer stem cells are stiffer, adhere better to the extracellular matrix (ECM), move slower both as single cells and clusters, display higher nuclear YAP, show a higher survival rate in response to mechanical confinement, and form larger transendothelial gaps. 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 single cell RNA-sequencing (scRNA-seq) expression patterns from a patient cohort, we show that this downregulation is a robust signature of colorectal tumors. Our results show that LGR5- cells display a mechanically dynamic phenotype suitable for dissemination from the primary tumor whereas LGR5+ cells display a mechanically stable and resilient phenotype suitable for extravasation and metastatic growth.

  View details for DOI 10.1038/s41467-024-47227-2

  View details for PubMedID 38637494

• CRISPR Knock-Ins in Organoids to Track Tumor Cell Subpopulations. Methods in molecular biology (Clifton, N.J.) Cortina, C., Canellas-Socias, A. 2024; 2811: 137-154

  Abstract

  The integration of CRISPR/Cas9 genome editing techniques with organoid technology has revolutionized the field of tumor modeling, enabling the creation of diverse tumor models with distinct mutational profiles. This protocol details the application of CRISPR knock-ins to engineer tumor organoids with reporter cassettes, which are regulated by endogenous promoters of specific genes of interest. This approach facilitates the precise fluorescent labeling, isolation, and subsequent manipulation of targeted tumor cell subpopulations. The utilization of these knock-in reporter cassettes not only allows the visualization and purification of specific tumor cell subsets but also enables conditional cell ablation and lineage tracing studies. In this chapter, we provide a comprehensive guide for the design, construction, delivery, and validation of CRISPR/Cas9 tools tailored for knock-in reporter cassette integration into specific marker genes of interest. By following this protocol, researchers can harness the potential of engineered tumor organoids to decipher intricate tumor heterogeneity, track metastatic trajectories, and unveil novel therapeutic vulnerabilities linked to specific tumor cell subpopulations.

  View details for DOI 10.1007/978-1-0716-3882-8_10

  View details for PubMedID 39037655

• Antitumor T-cell function requires CPEB4-mediated adaptation to chronic endoplasmic reticulum stress. The EMBO journal Fernandez-Alfara, M., Sibilio, A., Martin, J., Tusquets Uxo, E., Malumbres, M., Alcalde, V., Chanes, V., Canellas-Socias, A., Palomo-Ponce, S., Batlle, E., Mendez, R. 2023: e111494

  Abstract

  Tumor growth is influenced by a complex network of interactions between multiple cell types in the tumor microenvironment (TME). These constrained conditions trigger the endoplasmic reticulum (ER) stress response, which extensively reprograms mRNA translation. When uncontrolled over time, chronic ER stress impairs the antitumor effector function of CD8 T lymphocytes. How cells promote adaptation to chronic stress in the TME without the detrimental effects of the terminal unfolded protein response (UPR) is unknown. Here, we find that, in effector CD8 T lymphocytes, RNA-binding protein CPEB4 constitutes a new branch of the UPR that allows cells to adapt to sustained ER stress, yet remains decoupled from the terminal UPR. ER stress, induced during CD8 T-cell activation and effector function, triggers CPEB4 expression. CPEB4 then mediates chronic stress adaptation to maintain cellular fitness, allowing effector molecule production and cytotoxic activity. Accordingly, this branch of the UPR is required for the antitumor effector function of T lymphocytes, and its disruption in these cells exacerbates tumor growth.

  View details for DOI 10.15252/embj.2022111494

  View details for PubMedID 36919984

• Long-term platinum-based drug accumulation in cancer-associated fibroblasts promotes colorectal cancer progression and resistance to therapy. Nature communications Linares, J., Sallent-Aragay, A., Badia-Ramentol, J., Recort-Bascuas, A., Méndez, A., Manero-Rupérez, N., Re, D. L., Rivas, E. I., Guiu, M., Zwick, M., Iglesias, M., Martinez-Ciarpaglini, C., Tarazona, N., Varese, M., Hernando-Momblona, X., Cañellas-Socias, A., Orrillo, M., Garrido, M., Saoudi, N., Elez, E., Navarro, P., Tabernero, J., Gomis, R. R., Batlle, E., Pisonero, J., Cervantes, A., Montagut, C., Calon, A. 2023; 14 (1): 746

  Abstract

  A substantial proportion of cancer patients do not benefit from platinum-based chemotherapy (CT) due to the emergence of drug resistance. Here, we apply elemental imaging to the mapping of CT biodistribution after therapy in residual colorectal cancer and achieve a comprehensive analysis of the genetic program induced by oxaliplatin-based CT in the tumor microenvironment. We show that oxaliplatin is largely retained by cancer-associated fibroblasts (CAFs) long time after the treatment ceased. We determine that CT accumulation in CAFs intensifies TGF-beta activity, leading to the production of multiple factors enhancing cancer aggressiveness. We establish periostin as a stromal marker of chemotherapeutic activity intrinsically upregulated in consensus molecular subtype 4 (CMS4) tumors and highly expressed before and/or after treatment in patients unresponsive to therapy. Collectively, our study underscores the ability of CT-retaining CAFs to support cancer progression and resistance to treatment.

  View details for DOI 10.1038/s41467-023-36334-1

  View details for PubMedID 36765091

  View details for PubMedCentralID PMC9918738

• Cohesin couples transcriptional bursting probabilities of inducible enhancers and promoters NATURE COMMUNICATIONS Robles-Rebollo, I., Cuartero, S., Canellas-Socias, A., Wells, S., Karimi, M. M., Mereu, E., Chivu, A. G., Heyn, H., Whilding, C., Dormann, D., Marguerat, S., Rioja, I., Prinjha, R. K., Stumpf, M. H., Fisher, A. G., Merkenschlager, M. 2022; 13 (1): 4342

  Abstract

  Innate immune responses rely on inducible gene expression programmes which, in contrast to steady-state transcription, are highly dependent on cohesin. Here we address transcriptional parameters underlying this cohesin-dependence by single-molecule RNA-FISH and single-cell RNA-sequencing. We show that inducible innate immune genes are regulated predominantly by an increase in the probability of active transcription, and that probabilities of enhancer and promoter transcription are coordinated. Cohesin has no major impact on the fraction of transcribed inducible enhancers, or the number of mature mRNAs produced per transcribing cell. Cohesin is, however, required for coupling the probabilities of enhancer and promoter transcription. Enhancer-promoter coupling may not be explained by spatial proximity alone, and at the model locus Il12b can be disrupted by selective inhibition of the cohesinopathy-associated BET bromodomain BD2. Our data identify discrete steps in enhancer-mediated inducible gene expression that differ in cohesin-dependence, and suggest that cohesin and BD2 may act on shared pathways.

  View details for DOI 10.1038/s41467-022-31192-9

  View details for Web of Science ID 000831732000010

  View details for PubMedID 35896525

  View details for PubMedCentralID PMC9329429

• Mex3a marks drug-tolerant persister colorectal cancer cells that mediate relapse after chemotherapy NATURE CANCER Alvarez-Varela, A., Novellasdemunt, L., Barriga, F. M., Hernando-Momblona, X., Canellas-Socias, A., Cano-Crespo, S., Sevillano, M., Cortina, C., Stork, D., Morral, C., Turon, G., Slebe, F., Jimenez-Gracia, L., Caratu, G., Jung, P., Stassi, G., Heyn, H., Tauriello, D. F., Mateo, L., Tejpar, S., Sancho, E., Stephan-Otto Attolini, C., Batlle, E. 2022; 3 (9): 1052-+

  Abstract

  Colorectal cancer (CRC) patient-derived organoids predict responses to chemotherapy. Here we used them to investigate relapse after treatment. Patient-derived organoids expand from highly proliferative LGR5+ tumor cells; however, we discovered that lack of optimal growth conditions specifies a latent LGR5+ cell state. This cell population expressed the gene MEX3A, is chemoresistant and regenerated the organoid culture after treatment. In CRC mouse models, Mex3a+ cells contributed marginally to metastatic outgrowth; however, after chemotherapy, Mex3a+ cells produced large cell clones that regenerated the disease. Lineage-tracing analysis showed that persister Mex3a+ cells downregulate the WNT/stem cell gene program immediately after chemotherapy and adopt a transient state reminiscent to that of YAP+ fetal intestinal progenitors. In contrast, Mex3a-deficient cells differentiated toward a goblet cell-like phenotype and were unable to resist chemotherapy. Our findings reveal that adaptation of cancer stem cells to suboptimal niche environments protects them from chemotherapy and identify a candidate cell of origin of relapse after treatment in CRC.

  View details for DOI 10.1038/s43018-022-00402-0

  View details for Web of Science ID 000819258100001

  View details for PubMedID 35773527

  View details for PubMedCentralID 8670073

• Functional patient-derived organoid screenings identify MCLA-158 as a therapeutic EGFR x LGR5 bispecific antibody with efficacy in epithelial tumors NATURE CANCER Herpers, B., Eppink, B., James, M., Cortina, C., Canellas-Socias, A., Boj, S. F., Hernando-Momblona, X., Glodzik, D., Roovers, R. C., van de Wetering, M., Bartelink-Clements, C., Zondag-van der Zande, V., Mateos, J., Yan, K., Salinaro, L., Basmeleh, A., Fatrai, S., Maussang, D., van Bueren, J., Chicote, I., Serna, G., Cabellos, L., Ramirez, L., Nuciforo, P., Salazar, R., Santos, C., Villanueva, A., Attolini, C., Sancho, E., Palmer, H. G., Tabernero, J., Stratton, M. R., de Kruif, J., Logtenberg, T., Clevers, H., Price, L. S., Vries, R. J., Batlle, E., Throsby, M. 2022; 3 (4): 418-+

  Abstract

  Patient-derived organoids (PDOs) recapitulate tumor architecture, contain cancer stem cells and have predictive value supporting personalized medicine. Here we describe a large-scale functional screen of dual-targeting bispecific antibodies (bAbs) on a heterogeneous colorectal cancer PDO biobank and paired healthy colonic mucosa samples. More than 500 therapeutic bAbs generated against Wingless-related integration site (WNT) and receptor tyrosine kinase (RTK) targets were functionally evaluated by high-content imaging to capture the complexity of PDO responses. Our drug discovery strategy resulted in the generation of MCLA-158, a bAb that specifically triggers epidermal growth factor receptor degradation in leucine-rich repeat-containing G-protein-coupled receptor 5-positive (LGR5+) cancer stem cells but shows minimal toxicity toward healthy LGR5+ colon stem cells. MCLA-158 exhibits therapeutic properties such as growth inhibition of KRAS-mutant colorectal cancers, blockade of metastasis initiation and suppression of tumor outgrowth in preclinical models for several epithelial cancer types.

  View details for DOI 10.1038/s43018-022-00359-0

  View details for Web of Science ID 000787138500001

  View details for PubMedID 35469014

  View details for PubMedCentralID 2533206

• ERK1/2 Signaling Induces Upregulation of ANGPT2 and CXCR4 to Mediate Liver Metastasis in Colon Cancer CANCER RESEARCH Urosevic, J., Teresa Blasco, M., Llorente, A., Bellmunt, A., Berenguer-Llergo, A., Guiu, M., Canellas, A., Fernandez, E., Burkov, I., Clapes, M., Cartana, M., Figueras-Puig, C., Batlle, E., Nebreda, A. R., Gomis, R. R. 2020; 80 (21): 4668-4680

  Abstract

  Carcinoma development in colorectal cancer is driven by genetic alterations in numerous signaling pathways. Alterations in the RAS-ERK1/2 pathway are associated with the shortest overall survival for patients after diagnosis of colorectal cancer metastatic disease, yet how RAS-ERK signaling regulates colorectal cancer metastasis remains unknown. In this study, we used an unbiased screening approach based on selection of highly liver metastatic colorectal cancer cells in vivo to determine genes associated with metastasis. From this, an ERK1/2-controlled metastatic gene set (EMGS) was defined. EMGS was associated with increased recurrence and reduced survival in patients with colorectal cancer tumors. Higher levels of EMGS expression were detected in the colorectal cancer subsets consensus molecular subtype (CMS)1 and CMS4. ANGPT2 and CXCR4, two genes within the EMGS, were subjected to gain-of-function and loss-of-function studies in several colorectal cancer cell lines and then tested in clinical samples. The RAS-ERK1/2 axis controlled expression of the cytokine ANGPT2 and the cytokine receptor CXCR4 in colorectal cancer cells, which facilitated development of liver but not lung metastases, suggesting that ANGPT2 and CXCR4 are important for metastatic outgrowth in the liver. CXCR4 controlled the expression of cytokines IL10 and CXCL1, providing evidence for a causal role of IL10 in supporting liver colonization. In summary, these studies demonstrate that amplification of ERK1/2 signaling in KRAS-mutated colorectal cancer cells affects the cytokine milieu of the tumors, possibly affecting tumor-stroma interactions and favoring liver metastasis formation. SIGNIFICANCE: These findings identify amplified ERK1/2 signaling in KRAS-mutated colorectal cancer cells as a driver of tumor-stroma interactions that favor formation of metastases in the liver.

  View details for DOI 10.1158/0008-5472.CAN-19-4028

  View details for Web of Science ID 000587912700009

  View details for PubMedID 32816905