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Management of hepatoblastoma (HB), the most frequent pediatric liver cancer, is based on surgical resection and perioperative chemotherapy regimens. In this study, we aimed to identify actionable targets in HB and assess the efficacy of molecular therapies in preclinical models of HB. Paired tumor and adjacent tissues from 31 HBs and a validation set of 50 HBs were analyzed using RNA-seq, SNP, and methylation arrays. IGF2 overexpression was identified as the top targetable HB driver, present in 71% of HBs (22/31). IGF2high tumors displayed progenitor cell features and shorter recurrence-free survival. IGF2 overexpression was associated in 91% of cases with fetal promoter hypomethylation, ICR1 deregulation, 11p15.5 loss of heterozygosity or miR483-5p overexpression. The antitumor effect of xentuzumab (a monoclonal antibody targeting IGF1/2) alone or in combination with the conventional therapeutic agent cisplatin was assessed in HB cell lines, in PDX-derived HB organoids and in a xenograft HB murine model. The combination of xentuzumab with cisplatin showed strong synergistic antitumor effects in organoids and in IGF2high cell lines. In mice (n = 55), the combination induced a significant decrease in tumor volume and improved survival compared with cisplatin alone. These results suggest that IGF2 is an HB actionable driver and that, in preclinical models of HB, the combination of IGF1/2 inhibition with cisplatin induces superior antitumor effects than cisplatin monotherapy. Overall, our study provides a rationale for testing IGF2 inhibitors in combination with cisplatin in HB patients with IGF2 overexpression.

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Management of hepatoblastoma (HB), the most frequent pediatric liver cancer, is based on surgical resection and perioperative chemotherapy regimens. In this study, we aimed to identify actionable targets in HB and assess the efficacy of molecular therapies in preclinical models of HB. Paired tumor and adjacent tissues from 31 HBs and a validation set of 50 HBs were analyzed using RNA-seq, SNP, and methylation arrays. IGF2 overexpression was identified as the top targetable HB driver, present in 71% of HBs (22/31). IGF2high tumors displayed progenitor cell features and shorter recurrence-free survival. IGF2 overexpression was associated in 91% of cases with fetal promoter hypomethylation, ICR1 deregulation, 11p15.5 loss of heterozygosity or miR483-5p overexpression. The antitumor effect of xentuzumab (a monoclonal antibody targeting IGF1/2) alone or in combination with the conventional therapeutic agent cisplatin was assessed in HB cell lines, in PDX-derived HB organoids and in a xenograft HB murine model. The combination of xentuzumab with cisplatin showed strong synergistic antitumor effects in organoids and in IGF2high cell lines. In mice (n = 55), the combination induced a significant decrease in tumor volume and improved survival compared with cisplatin alone. These results suggest that IGF2 is an HB actionable driver and that, in preclinical models of HB, the combination of IGF1/2 inhibition with cisplatin induces superior antitumor effects than cisplatin monotherapy. Overall, our study provides a rationale for testing IGF2 inhibitors in combination with cisplatin in HB patients with IGF2 overexpression.

At present, we can understand maps as a result of multiple information layers about places. Today, the new media are capable of dimensioning, registering and locating practically every piece of ground of our planet. However, the need to map emotional territories about our subjective experiences in the landscape is motivating the creation of new cartographic proposals though art. Walter Benjamin pointed out that art is capable of “combing history against the grain”. In our historical context, characterized by the use of topographic measurement and identification tools, it becomes essential to allow discontinuous things to appear and claiming transversal tools with which to understand our world, question it, complement it and even escape from it.; Podemos entender los mapas como la suma de múltiples capas de información precisa y objetiva que presentan las características de un lugar y que han ido ampliándose y perfeccionándose a lo largo de la historia. A día de hoy, las nuevas tecnologías han conseguido medir, registrar y localizar prácticamente cada parcela de nuestro planeta. No obstante, la necesidad de cartografiar territorios emocionales a partir de la subjetividad de nuestra experiencia en el paisaje está generando la creación de nuevas propuestas cartográficas por parte de los artistas. Walter Benjamin apuntaba que el arte es capaz de peinar la historia a contrapelo. En el contexto histórico que nos ocupa, en el que la localización territorial y la identificación topográfica, en cierto sentido, pierden importancia en favor de la interacción, cobra importancia el acto de “dejar aparecer lo discontinuo” y reivindicar herramientas transversales con las que entender nuestro mundo, cuestionarlo, complementarlo e incluso escapar de él.

El recurso se enmarca dentro del Proyecto de Innovación Docente de la Facultad de Bellas Artes “Modelos didácticos audiovisuales como soporte fundamental a la actividad docente, a través de la Plataforma de Enseñanza Virtual en los Grados de Bellas Artes y Conservación y Restauración de Bienes Culturales”, el cual está destinado a la creación de material para el apoyo a la formación dentro del Tercer Plan Propio de Docencia de la Universidad de Sevilla. En este caso, el trabajo realizado desde el Dpto. de Pintura recoge en un video breve dos procesos completos de elaboración de una obra pictórica realizados mediante dos metodologías distintas. Este video dirigido al alumnado de la Facultad de Bellas Artes, está especialmente ideado para alumnado de primer curso de los grados en Bellas Artes y Conservación y restauración de bienes culturales. Pero al ser un recurso de carácter procedimental y abordar conceptos que se usan a lo largo de todo el periodo de formación, resulta de gran utilidad como material de consulta durante cualquiera de los cursos, pues es una temática de índole práctica que se imparte solo a comienzos de ambos grados. El recurso producido aborda diferentes maneras de comenzar a trabajar con la materia pictórica, concretamente los métodos de “pintura directa “ y “pintura tonal”. Dividido en dos partes, el primer proceso de intervención realizado por el Pr. García Perera nos muestra cómo partiendo de una valoración general de la composición, la obra va trabajándose de lo general a lo particular mediante una paleta de color diferenciada. Mientras que, en la segunda parte, nuestro antiguo compañero el Pr. Antonio Agudo hace una colaboración mostrando cómo el método de pintura tonal parte de un núcleo cromático neutro que posteriormente se matiza y valora hasta llegar a la conclusión final de la obra.

BACKGROUND: Polymerized allergens conjugated to non-oxidized mannan (PM-allergoids) are novel vaccines targeting dendritic cells (DCs). Previous experimental data indicate that PM-allergoids are readily taken up by DCs and induce Treg cells. This first-in-human study was aimed to evaluate safety and to find the optimal dose of house dust mite PM-allergoid (PM-HDM) administered subcutaneously (SC) or sublingually (SL). METHODS: In a randomized, double-blind, double-dummy, placebo-controlled trial, 196 subjects received placebo or PM-HDM at 500, 1000, 3000, or 5000 mannan-conjugated therapeutic units (mTU)/mL in 9-arm groups for 4 months. All subjects received 5 SC doses (0.5 ml each) every 30 days plus 0.2 ml SL daily. The primary efficacy outcome was the improvement of titrated nasal provocation tests (NPT) with D. pteronyssinus at baseline and at the end of the study. All adverse events and reactions were recorded and assessed. Secondary outcomes were the combination of symptom and medication scores (CSMS) and serological markers. RESULTS: No moderate or severe adverse reactions were reported. Subjects improving the NPT after treatment ranged from 45% to 62% in active SC, 44% to 61% in active SL and 16% in placebo groups. Statistical differences between placebo and active groups were all significant above 500 mTU, being the highest with 3000 mTU SL (p = 0.004) and 5000 mTU SC (p = 0.011). CSMS improvement over placebo reached 70% (p < 0.001) in active 3000 mTU SC and 40% (p = 0.015) in 5000 mTU SL groups. CONCLUSIONS: PM-HDM immunotherapy was safe and successful in achieving primary and secondary clinical outcomes in SC and SL at either 3000 or 5000 mTU/ml.

Background & aims: single-agent anti-PD1 checkpoint inhibitors convey outstanding clinical benefits in a small fraction (∼20%) of patients with advanced hepatocellular carcinoma (aHCC) but the molecular mechanisms determining response are unknown. To fill this gap, we herein analyze the molecular and immune traits of aHCC in patients treated with anti-PD1. Methods: overall, 111 tumor samples from patients with aHCC were obtained from 13 centers before systemic therapies. We performed molecular analysis and immune deconvolution using whole-genome expression data (n = 83), mutational analysis (n = 72), and histologic evaluation with an endpoint of objective response. Results: among 83 patients with transcriptomic data, 28 were treated in frontline, whereas 55 patients were treated after tyrosine kinase inhibitors (TKI) either in second or third line. Responders treated in frontline showed upregulated interferon-γ signaling and major histocompatibility complex II-related antigen presentation. We generated an 11-gene signature (IFNAP), capturing these molecular features, which predicts response and survival in patients treated with anti-PD1 in frontline. The signature was validated in a separate cohort of aHCC and >240 patients with other solid cancer types where it also predicted response and survival. Of note, the same signature was unable to predict response in archival tissue of patients treated with frontline TKIs, highlighting the need for fresh biopsies before immunotherapy. Conclusion: interferon signaling and major histocompatibility complex-related genes are key molecular features of HCCs responding to anti-PD1. A novel 11-gene signature predicts response in frontline aHCC, but not in patients pretreated with TKIs. These results must be confirmed in prospective studies and highlights the need for biopsies before immunotherapy to identify biomarkers of response.

Background: Transforming growth factor β (TGFβ) has a dual role in cancer, including hepatocellular carcinoma (HCC). It acts as a tumor suppressor in early stages of hepatocarcinogenesis, but promotes epithelial-to-mesenchymal transition, angiogenesis and immunosuppression in advanced stages. PMEPA1 (prostate transmembrane protein androgen induced 1) negatively regulates TGFβ signaling by interacting with SMAD proteins, and has been shown to promote TGFβ oncogenic effects in other cancers. Thus, we aimed to explore the role of PMEPA1 in HCC pathogenesis. Methods: We analyzed transcriptomic, genomic, epigenomic and clinicopathological data of a discovery cohort comprising 228 HCCs and a validation cohort of 361 HCCs. PMEPA1 levels were quantified by qPCR in the discovery cohort. PMEPA1 overexpression was validated in 6 independent publicly available cohorts (n = 916) with either microarray or RNAseq data. We also evaluated PMEPA1 expression at the single-cell level using a publicly available single-cell RNA sequencing (scRNAseq) dataset comprising 22 HCC samples. Genetically engineered mouse models were generated by performing hydrodynamic tail-vein injection of equal amounts of plasmids overexpressing MYC and MYC+PMEPA1. Molecular and histopathological analyses of the murine tumors are currently ongoing. Results: PMEPA1 was overexpressed in 18% of human HCCs [FC > 2; n = 203/1144], a feature associated with TGFβ signaling (p < 0.05). Overexpression of PMEPA1 was associated with gene body hypermethylation (p < 0.0001). HCCs displaying both TGFβ signaling and high PMEPA1 levels (11% of cases) were significantly enriched in signatures of immune exhaustion, late TGFβ activation, tumor microenvironment (TME) response to TGFβ and active stroma (p < 0.05) when compared with HCCs with TGFβ signalling alone (8% of cases) or PMEPA1 overexpression alone (11% of cases). Analyses of PMEPA1 expression at the single-cell level revealed that this gene is not only expressed by tumoral cells but also by the TME cell populations, including endothelial cells, pericytes, fibroblasts, and dendritic cells. In vivo, overexpression of MYC+PMEPA1 led to HCC development in 9/16 mice and was associated with a significantly reduced survival when compared to mice overexpressing MYC alone (p = 0.014), which did not develop tumors. Compared to healthy liver controls, MYC+PMEPA1 tumors presented upregulation of AKT. Conclusion: PMEPA1 upregulation is linked to TGF-β activation and an aggressive phenotype in human HCC. Overexpression of PMEPA1 in combination with MYC in vivo led to HCC development, showing for the first time the oncogenic role of PMEPA1 in this cancer type. Citation Format: Marta Piqué-Gili, Carmen Andreu-Oller, Roser Pinyol, Roger Esteban, Marina Bárcena-Varela, Judit Peix, Jordi Abril-Fornaguera, Katherine E. Lindblad, Miguel Torres-Martin, Júlia Huguet-Pradell, Daniela Sia, Amaia Lujambio, Josep M Llovet. PMEPA1 has an oncogenic role in the context of TGF-β signaling in hepatocellular carcinoma [abstract]. In: Proceedings of the AACR Special Conference: Advances in the Pathogenesis and Molecular Therapies of Liver Cancer; 2022 May 5-8; Boston, MA. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(17_Suppl):Abstract nr PR02.

Background: Transforming growth factor β (TGFβ) has a dual role in cancer, including hepatocellular carcinoma (HCC). It acts as a tumor suppressor in early stages of hepatocarcinogenesis, but promotes epithelial-to-mesenchymal transition, angiogenesis and immunosuppression in advanced stages. PMEPA1 (prostate transmembrane protein androgen induced 1), a direct target gene of the TGFβ pathway that negatively regulates TGFβ signaling by interacting with SMAD proteins, has been shown to promote TGFβ oncogenic effects in other cancers and we decided to explore its role in HCC pathogenesis. Methods: We analyzed transcriptomic, genomic, epigenomic and clinicopathological data of a discovery cohort of 228 HCCs and a validation cohort of 361 HCCs. PMEPA1 levels were quantified by qPCR in the discovery cohort. PMEPA1 overexpression was validated in 6 independent cohorts (n = 916) using microarray and RNAseq data. Genetically engineered mouse models were generated through hydrodynamic tail-vein injection. Equal amounts of plasmids overexpressing PMEPA1, TGFB1 and MYC were injected in mice to evaluate survival. Molecular and histopathological analyses of the murine tumors are currently ongoing. Results: PMEPA1 was overexpressed in 18% of human HCCs [FC > 2; n = 203/1144], a feature associated with TGFβ signaling (p < 0.05). Overexpression of PMEPA1 was associated with gene body hypermethylation (p < 0.0001). HCCs displaying both TGFβ signaling and high PMEPA1 levels (11% of cases) were significantly enriched in signatures of immune exhaustion, late TGFβ activation, TME response to TGFβ and active stroma (p < 0.05) compared to HCCs with TGFβ signalling alone (8% of cases) or PMEPA1 high levels alone (11% of cases). In vivo, overexpression of MYC+PMEPA1 led to HCC development in 9/16 mice and showed a lower survival when compared to the control arm, overexpression of MYC alone (p = 0.014). On the contrary, only 1/12 mice in the MYC+PMEPA1+TGFβ1 condition developed HCC and none of the MYC+TGFβ1 mice. Conclusion: PMEPA1 upregulation is linked to TGF-β activation and an aggressive phenotype in human HCC. Overexpression PMEPA1 in combination with MYC in vivo led to HCC development, showing for the first time the oncogenic role of PMEPA1 in this cancer. Citation Format: Marta Piqué-Gili, Carmen Andreu-Oller, Roser Pinyol, Marina Bárcena-Varela, Roger Esteban-Fabró, Judit Peix, Jordi Abril-Fornaguera, Katherine E. Lindblad, Miguel Torres-Martin, Daniela Sia, Amaia Lujambio, Josep M. Llovet. PMEPA1 is an oncogene linked to TGF-β signaling in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 840.

BACKGROUND AND AIMS Non-alcoholic steatohepatitis (NASH)-related hepatocellular carcinoma (HCC) is increasing globally, but its molecular features are not well defined. We aimed to identify unique molecular traits characterising NASH-HCC compared to other HCC aetiologies. METHODS We collected 80 NASH-HCC and 125 NASH samples from 5 institutions. Expression array (n = 53 NASH-HCC; n = 74 NASH) and whole exome sequencing (n = 52 NASH-HCC) data were compared to HCCs of other aetiologies (n = 184). Three NASH-HCC mouse models were analysed by RNA-seq/expression-array (n = 20). Activin A receptor type 2A (ACVR2A) was silenced in HCC cells and proliferation assessed by colorimetric and colony formation assays. RESULTS Mutational profiling of NASH-HCC tumours revealed TERT promoter (56%), CTNNB1 (28%), TP53 (18%) and ACVR2A (10%) as the most frequently mutated genes. ACVR2A mutation rates were higher in NASH-HCC than in other HCC aetiologies (10% vs. 3%, p

Aim: To evaluate tolerability and efficacy of Parietaria judaica subcutaneous immunotherapy on patients with allergic rhinoconjunctivitis. Patients & methods: 51 patients were assigned to build-up scheme (six increasing doses) of P. judaica depot native extract, plus three maintenance monthly administrations. Results: Out of 470 administered doses, only 3.8% elicited systemic reactions (1.5% nonspecific and 2.3% grade I). Concerning the exploratory efficacy parameters: cutaneous reactivity at the final visit versus baseline was significantly decreased; specific titers of IgG and IgG4 increased significantly and patients showed a significant decrease in the rhinitis symptoms score. Conclusion: P. judaica subcutaneous immunotherapy (Allergovac® depot ROXALL Medicina España S.A., Zamudio, Spain) with an abbreviated up-dosing scheme showed an adequate safety and tolerability profile and induced preliminary efficacy changes.

El olivar es el cultivo mediterráneo por excelencia, donde destaca tanto por su extensión como por su importancia milenaria. La cultura que se le asocia hunde sus raíces en los primeros procesos de domesticación agraria y ha tenido un papel fundamental en la ecología, la economía, la historia y la belleza del Mediterráneo y sus paisajes. Este artículo tiene como objetivo fundamental abordar desde una perspectiva artística, interdisciplinar y transdiciplinar los valores del paisaje del olivar andaluz, analizando algunas de las formas objetivas que ha adoptado en distintas áreas del territorio y profundizando en las percepciones y las emociones que estas suscitan. Partimos del estudio interdisciplinar de los valores de los paisajes de olivar para destacar su riqueza antropológica y estética, como patrimonio natural y cultural, sin olvidar algunos de los principales problemas ambientales que está suponiendo para estos las exigencias crecientes del mercado que lleva en numerosas ocasiones a la adopción de insumos y manejos insostenibles en el tratamiento del olivo en menoscabo de la riqueza de estos paisajes. Damos un paso metodológico más desarrollando trabajos de carácter transdisciplinar que abordan el paisaje del olivar creando relatos más allá del contexto disciplinar de los participantes, The olive grove is the pre-eminent crop in the Mediterranean Sea, where it stands out due the extension of the crops and its millenary importance. The olive culture is rooted in the very first processes of agrarian domestication, and it played a major role in the ecology, economy, history and beauty of the Mediterranean sea and its landscapes. The main goal of this article is to address the value of the Andalusian, olive-growing landscapes from an artistic, interdisciplinar and transactional perspective by analysing some of the objective shapes that they have adopted in various areas of the territory and delving into the perceptions and emotions that they arouses. We start with the interdisciplinar study of the values of the olive-growing landscapes to emphasise their anthropologic and aesthetic wealth as a natural and cultural heritage, acknowledging some of the major environmental problems that are taking place, consequence of the increase in market demand that leads to the extraction of supplies and the use of unsustainable control treatments at the expense of the wealth of these landscapes. We go further on the methodology by developing interdisciplinary works that would address the olive-growing landscapes, creating stories beyond the disciplinar context of the participants

El 27 de enero de 2017, la UNESCO ha incluido al paisaje del olivar andaluz en una lista tentativa de futuros patrimonios mundiales, lo que presenta oficialmente en el mundo estos paisajes y exige mantener sus valores patrimoniales. Aquí se ofrece una lectura científica e interdisciplinar de las claves geográficas, históricas y estéticas que justifican tal consideración y también se presenta la problemática ambiental y social que está suponiendo para aquellos seculares paisajes la adopción de insumos y manejos insostenibles del olivo para adaptar sus producciones a un exigente mercado.

Background Allergy to olive pollen is one of the primary causes of allergic asthma in Spain. Even though allergen immunotherapy (AIT) has shown clinical benefits in patients sensitized to different allergens, studies in asthmatic patients sensitized to olive pollen are insufficient. Objective To assess the effectiveness and safety of an ultra-short course of AIT with an L-tyrosine-adsorbed and monophosphoryl lipid A-adjuvanted olive pollen and olive/grass pollen extract (Pollinex Quattro®) in patients with allergic asthma in the real-world setting. Methods Retrospective, controlled study including patients with asthma, with and without allergic rhinitis, caused by sensitization to olive pollen from 11 centers in Spain. Patients received out-of-season (October–March) treatment with AIT in addition to their pharmacological treatment (active group) or pharmacological treatment (control group). Effectiveness variables, including unscheduled visits to the healthcare center, emergency room admissions, symptoms of asthma and rhinitis (following GEMA and ARIA classifications, respectively), and use of medication to treat asthma and rhinitis during the subsequent pollen season were compared between treatment groups. Results Of 131 study patients, 42 were treated with their usual asthma medication (control group) and 89 were treated with AIT (active group), either Pollinex Quattro® 100% olive pollen (n = 43, 48.3%) or 50% olive pollen/50% grass pollen (n = 46, 51.7%). Patients’ demographic and clinical characteristics were similar between groups. The mean (SD) number of unscheduled visits to a healthcare center and emergency room admissions due to allergy symptoms was 2.19 (1.40) and 0.43 (0.63) in the control group, and 1.09 (1.25) and 0.11 (0.51) in the active group (P = 0.001 and P = 0.006, respectively). Severity and control of asthma symptoms remained unchanged (P = 0.347 and P = 0.179, respectively), rhinitis type improved (P = 0.025), and severity remained unchanged in the active compared to the control group. The use of short-acting beta-agonists and inhaled corticosteroids to treat asthma symptoms decreased in the active vs. the control group (P = 0.001 and P = 0.031, respectively). Twelve (13.5%) and two (2.2%) patients in the active group experienced local adverse reactions (edema, swelling, erythema, hives, pruritus, and heat), and systemic adverse reactions (hypertensive crisis and low-grade fever) to AIT, respectively; none was serious. Conclusion AIT with Pollinex Quattro® specific for olive pollen and olive/grass pollens resulted in reduced visits to the healthcare center and emergency room and the use of asthma medication during the pollen season, indicating that this treatment was safe and effective in treating asthma in patients sensitized to these pollens.

Background Tropomyosin is the most studied shellfish allergen and has been involved in cross-reactivity among different invertebrates (crustacean, mollusks, mites, insects, and nematodes). Objective To determine the relevance of tropomyosin in mite- and shellfish-sensitized patients using tropomyosin skin testing. Methods Patients were divided into 3 groups: group M included mite allergic patients (ie, individuals with respiratory symptoms and a positive result on skin prick testing [SPT] to house dust mites), group S included shellfish allergic patients (ie, individuals who reported symptoms with shellfish), and group MS included mite- and shellfish allergic patients (ie, individuals who simultaneously fulfilled the inclusion criteria for groups M and S). Tropomyosin was purified from shrimp, characterized, and used in SPT for diagnosis in the patient population. Results Eight hundred fifty patients were included in the study: 790 (92.9%) in group M, 21 (2.5%) in group S, and 39 (4.6%) in group MS. Tropomyosin was purified from shrimp with a purity higher than 95%. Forty-two individuals tested positive to tropomyosin: the prevalence was 2.7% in group M, 28.6% in group S, and 38.5% in patients of group MS. Twenty-one (50%) of the tropomyosin-positive individuals had symptoms with shellfish, and 3 (14.3%) reported anaphylaxis. Conclusion The prevalence of tropomyosin was low in mite-sensitized patients (2.7 %) and high in shellfish allergic patients (28.6%). The higher prevalence of tropomyosin was found in patients sensitized to both mite and shellfish (38.5%). The selection of tropomyosin-sensitized patients by SPT might help in the choice of appropriate treatments or management for these patients.

Background and aim: Non-alcoholic steatohepatitis (NASH) is emerging as one of the leading risk factors for hepatocellular carcinoma (HCC), but its molecular pathogenesis is still ill-defined. This study aims to identify unique molecular traits that differentiate NASH-HCC from other aetiologies through an integrative molecular characterization. Methods: A total of 225 tissue samples were collected, including samples from 125 biopsied/transplanted NASH patients; and 100 resected/transplanted NASH-HCC patients. Molecular characterization of FFPE samples, comprised expression array (n=53 NASH-HCC; n=74 NASH), whole exome sequencing (n=50 NASH-HCC), and SNP array (n=44 NASH-HCC). Publicly available data from HCV/HBV/alcohol-related HCCs were used to identify NASH-HCC distinctive traits. Results: NASH-HCC patients compared to NASH non-HCC patients were prevalently males (82% vs 42%, p Conclusions: Non-tumour liver tissue of NASH-HCC patients is characterized by a cancer-field enriched in inflammatory, epithelial-to-mesenchymal transition and proliferation signalling pathways. NASH-HCC tumours showed a high frequency of ACVR2A mutations (10%), and a novel cancer mutational signature #3 (15%), suggesting genotoxic factors specifically associated to this entity. Citation Format: Sara Torrecilla, Roser Pinyol, Huan Wang, Carla Montironi, Carmen Andreu-Oller, Wei Qiang Leow, Agrin Moeini, Claudia Oliveira, Venancio Avancini Ferreira Alves, Anja Lachenmayer, Stephanie Roessler, Beatriz Minguez, Peter Schirmacher, Paolo Boffetta, Jean-François Dufour, Swan N Thung, Andrew Uzilov, Flair Jose Carrilho, Charissa Chang, Daniela Sia, Josep M Llovet. Distinctive molecular traits of hepatocellular carcinoma in patients with non-alcoholic steatohepatitis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 461.

Background: Although allergy to Cannabis sativa was first reported over 40 years ago, the allergenicity has scarcely been studied. The objectives of this study were to investigate the frequency of sensitization to this plant, to analyze the clinical characteristics and allergenic profile of sensitized individuals and to identify the allergens involved. Methods: Five hundred and forty-five individuals in Spain attending allergy clinics with respiratory or cutaneous symptoms underwent a skin-prick test (SPT) with C. sativa leaf extract. The extract was characterized by SDS-PAGE and 2-dimensional electrophoresis. Specific IgE to C. sativa was measured in positive SPT individuals. The clinical and allergenic profiles of sensitized individuals were investigated and the most-recognized allergens sequenced and characterized by liquid chromatography-mass spectrometry/mass spectrometry. Results: Of this preselected population, 44 individuals had positive SPT to C. sativa (prevalence 8.1%). Prevalence was higher in individuals who were C. sativa smokers (14.6%). Two individuals reported mild symptoms with C. sativa. Twenty-one individuals from 32 available sera (65.6%) had positive specific IgE to C. sativa. Twelve sera recognized at least 6 different bands in a molecular-weight range of between 10 and 60 kDa. Six of them recognized a 10-kDa band, identified as a lipid transfer protein (LTP) and 8 recognized a 38-kDa band, identified as a thaumatin-like protein. Conclusions: There is a high prevalence of sensitization to C. sativa leaves. The clinical symptoms directly attributed to C. sativa were uncommon and mild. The sensitization profile observed suggests that C. sativa sensitization may be mediated by two mechanisms, i.e. cross-reactivity, mainly with LTP and thaumatin-like protein, and exposure-related ‘de novo' sensitization.

Background Cross-reactivity among fruits and different pollen and fruit species has been extensively reported. Objectives To investigate the in vitro cross-reactivity between tomato and pollen, fruit, and latex extracts and to identify the proteins involved. Methods A serum pool was prepared from 18 individuals residing on the Spanish Mediterranean coast (9 men and 9 women; mean [SD] age, 27.4 [10.1] years) who had positive skin prick test reactions to tomato peel. Extracts from 10 pollens, 12 fruits, and latex were tested. Levels of specific IgE to each extract were measured. The allergenic profile was evaluated by means of immunoblot. The percentage of inhibition between extracts and tomato peel extract was analyzed by means of CAP inhibition, and the allergens implicated were elucidated by immunoblot inhibition. Results For pollens, the highest specific IgE values were obtained for grasses. Most pollen extracts showed a capacity of inhibition similar to that of tomato peel extract; high percentages were obtained with Artemisia vulgaris and Poa pratensis . The most strongly inhibited allergens in tomato corresponded to bands of 32 and 45 kDa. For fruits, the highest value of specific IgE was detected for peach. High percentages of inhibition were obtained with peach and hazelnut. No inhibition was detected with latex. Peach, chestnut, and melon inhibited high molecular weight bands (32 and 45 kDa) and a band of approximately 10 kDa. Conclusions Cross-reactivity between tomato and pollen and fruit extracts has been demonstrated. Allergens with a high molecular weight range seem to be responsible in pollen extracts. A 10-kDa band seems to be responsible in Platanus acerifolia, Salsola kali , peach, chestnut, and melon.

Background: Sensitivity to Chenopodiaceae is a frequent cause of allergic respiratory diseases in geographic areas where sensitization to Salsola kali and Chenopodium album has been reported. The objective of this study was to evaluate the pattern of sensitization to 3 Salsola species in patients residing on the Mediterranean coast of south-eastern Spain. Methods:S. kali, S. vermiculata and S. oppositifolia pollen extracts were prepared. Patients reporting respiratory and/or cutaneous symptoms were skin prick tested with the 3 Salsola extracts. Individuals with positive skin prick tests to at least 1 of the 3 Salsola species were included. Specific IgE was determined by direct ELISA. SDS-PAGE and 2-D analysis were conducted to elucidate the protein profile. The allergenic profile was investigated by immunoblot. Inhibition experiments were conducted to establish cross-reactivity between different species. Results: 246 patients were included. 237 patients (96.3%) tested positive to S. oppositifolia, 189 (76.8%) to S. kali and 185 (75.2%) to S. vermiculata. Protein profile and immunoblot demonstrated similar patterns in all extracts, except in low-molecular-weight allergens of S. oppositifolia. Immunoblot inhibition experiments demonstrated that most high-molecular-weight allergens of S. oppositifolia were inhibited by S. kali whereas low-molecular-weight allergens were totally inhibited only by C. album. Conclusions: This study confirms the allergenic importance of other Salsola species, especially S. oppositifolia. We have demonstrated that the 3 species show a high degree of cross-reactivity, but S. oppositifolia shares more allergenic similarities with C. album than S. kali.

Summary Background Specifically designed recombinant allergens with reduced IgE reactivity are promising candidates for a more defined, effective, and safer specific immunotherapy (SIT). Objective We sought to obtain hypoallergenic hybrid molecules which could potentially be applied to house dust mite (HDM) allergy treatment. Methods Two hybrid molecules (QM1 and QM2) derived from the two major Dermatophagoides pteronyssinus allergens, Der p 1 and Der p 2, were engineered by PCR, produced in Escherichia coli, and purified. The overall IgE-binding capacity of the hybrids was compared with their single components by Western blot, specific IgE, skin prick test (SPT), and IgE-inhibition assays. T cell proliferation assay were performed to confirm their retention of T cell reactivity. Immune responses to the hybrid molecules were studied in BALB/c mice. Results The IgE reactivity of both hybrid proteins was strongly reduced as evaluated by in vitro methods. Furthermore, in vivo SPTs performed on 106 HDM-allergic patients showed that the hybrid proteins had a significantly lower potency to induce cutaneous reactions than the individual components. Hybrid molecules induced higher T cell proliferation responses than those produced by an equimolecular mixture of Der p 1 and Der p 2. Immunization of mice with the hybrid proteins induced Der p 1- and Der p 2-specific IgG, which inhibited the binding of allergic patients' IgE to these natural allergens. Conclusion QM1 and QM2 hybrids exhibited less IgE-binding activity but preserved immunogenicity and fulfilled the basic requirements for hypoallergenic molecules suitable for a future SIT of HDM allergy.

Background Despite the use of conventional allergen-specific immunotherapy in clinical practice, more defined, efficient, and safer allergy vaccines are required. Objective The aim of the study was to obtain hypoallergenic molecules by deleting B-cell epitopes, which could potentially be applied to Parietaria judaica pollen allergy treatment. Methods Three hybrid molecules (Q1, Q2, and Q3) derived from fragments of the 2 major P judaica pollen allergens, Par j 1 and Par j 2, were engineered by means of PCR. Hybrid structures were compared with their natural components by means of circular dichroism, and their biologic activities were compared by using T-cell proliferation assays. Their IgE-binding activity was determined with Western blotting, skin prick tests, and enzyme allergosorbent and ELISA inhibition tests. Results The hybrid proteins, especially Q2 and Q3, revealed significantly reduced IgE reactivity compared with the natural allergens, as well as with the whole P judaica extract. Furthermore, in vivo skin prick tests showed that the hybrid proteins had a significantly lower potency to induce cutaneous reactions than the whole P judaica extract. Two (Q1 and Q2) of the 3 hybrid proteins induced a comparable T-cell proliferation response as that produced by the whole extract and natural allergens. Conclusion Considering its reduced anaphylactogenic potential, together with its conserved T-cell reactivity, the engineered Q2 protein could be used in safe and shortened schedules of allergen-specific immunotherapy against P judaica pollen allergy. Clinical implications Recombinant hybrid Q2 is able to induce T-cell proliferation, thus evidencing a potential therapeutic effect. Its reduced IgE-binding capacity envisages an excellent safety profile.

Background Seafood plays an important role in human nutrition and is responsible for severe hypersensitivity reactions. Objective To evaluate how the cooking process may alter the in vivo and in vitro allergenicity of these extracts. Methods Raw and boiled extracts of shrimps and 2 types of lobsters were manufactured. Boiled extracts were prepared after the raw material was boiled for 15 minutes in phosphate-buffered saline. Raw and boiled extracts were homogenized and extracted for 4 hours. Afterward, the extracts were centrifuged, dialyzed, filtered, and freeze-dried. Seventy-eight patients were skin prick tested with these raw and boiled extracts. Specific IgE against the 6 extracts was measured by enzyme-linked immunosorbent assay (ELISA). Immunoblots and ELISA inhibition studies were performed with a pool of sera. Results In vivo results showed that boiled extracts induced statistically significant larger wheals than raw extracts. More patients with positive results were also detected with boiled extracts. In vitro experiments by direct ELISA confirmed the in vivo results. The protein content in the boiled extracts decreased, and important differences were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Crossreactivity experiments showed that both types of extracts retained similar allergenic characteristics, even if the immunoblots revealed some differences in IgE binding. Conclusions More patients were identified using boiled extracts of shrimp and American and spiny lobsters than with raw extracts. The wheal sizes of the skin test reactions and specific IgE levels were also significantly greater using boiled extracts. The use of boiled extracts seems to be more effective in diagnosing seafood allergy.

Background The development of sensitization and symptoms after the inhalation of epithelial allergens is common. Objectives To investigate the allergic response (in vivo and in vitro) of 3 individuals clinically sensitive and occupationally exposed to cow and goat and to evaluate the allergenic cross-reactivity between cow and goat epithelium extracts. Methods Three patients—a butcher and 2 doctors in veterinary medicine—were evaluated. These patients reported allergic respiratory symptoms after occupational exposure to goats and cows. Extracts were prepared from epithelia of both animals. All the patients underwent organ-specific allergen challenges with cow and goat extracts. Four nasal and 2 bronchial challenges were conducted following standardized procedures. Results All 3 patients had positive challenge results with the offending allergen extract. Specific IgE to goat allergens was detected in all patients. Several IgE-binding bands were detected. Inhibition assays (enzyme allergosorbent test and immunoblots) confirmed moderate-to-high cross-reactivity between goat and cow extracts. Conclusions We confirm clinical sensitivity and specific IgE binding to goat and cow allergens in occupationally exposed individuals. There was good correlation among the clinical history, exposure, and the laboratory findings.

Three tomato allergens (Lyc e 1, Lyc e 2 and Lyc e 3) have been characterised until now. Previous studies have demonstrated the existence of other proteins with capacity to bind IgE. The objective of this study was to identify new tomato allergens by molecular biology techniques. A tomato peel cDNA library was screened using a specific pool of sera from sensitised individuals. A clone was obtained and its sequence compared to other described allergens. This clone contained 34 aminoacids from the C-terminal region of an acidic ribosomal protein 60S. This protein showed more than 85% of homology with fungal allergens and 93.8% of homology with the almond allergen Pru du 5. The clone also contains 169 base pairs without homology with other known genes and could correspond to non-coding sequences. The IgE detection with a pool of sera was positive which confirms that is a new tomato allergen.

Although allergy to Cannabis sativa was first reported over 40 years ago, the allergenicity has scarcely been studied. The objectives of this study were to investigate the frequency of sensitization to this plant, to analyze the clinical characteristics and allergenic profile of sensitized individuals and to identify the allergens involved.Five hundred and forty-five individuals in Spain attending allergy clinics with respiratory or cutaneous symptoms underwent a skin-prick test (SPT) with C. sativa leaf extract. The extract was characterized by SDS-PAGE and 2-dimensional electrophoresis. Specific IgE to C. sativa was measured in positive SPT individuals. The clinical and allergenic profiles of sensitized individuals were investigated and the most-recognized allergens sequenced and characterized by liquid chromatography-mass spectrometry/mass spectrometry.Of this preselected population, 44 individuals had positive SPT to C. sativa (prevalence 8.1%). Prevalence was higher in individuals who were C. sativa smokers (14.6%). Two individuals reported mild symptoms with C. sativa. Twenty-one individuals from 32 available sera (65.6%) had positive specific IgE to C. sativa. Twelve sera recognized at least 6 different bands in a molecular-weight range of between 10 and 60 kDa. Six of them recognized a 10-kDa band, identified as a lipid transfer protein (LTP) and 8 recognized a 38-kDa band, identified as a thaumatin-like protein.There is a high prevalence of sensitization to C. sativa leaves. The clinical symptoms directly attributed to C. sativa were uncommon and mild. The sensitization profile observed suggests that C. sativa sensitization may be mediated by two mechanisms, i.e. cross-reactivity, mainly with LTP and thaumatin-like protein, and exposure-related 'de novo' sensitization.

Background The prevalence of sensitization to tomato has been estimated in 6.5% in a Mediterranean population. Objectives To characterize the allergenic profile of 6 commonly ingested varieties of tomato and to identify and quantify the most representative allergens. Methods Rama, Rambo, Canario, Kumato, Pera and Raf tomato varieties were carefully peeled and extracts from peel and pulp prepared. The extracts were characterized by SDS-PAGE and immunoblot. Lyc e 3 and osmotin-like protein (OLP) were identified by LC MS/MS and PG2A by immunoblot with a specific polyclonal antibody. Protein content and allergen concentration (Lyc e 3 and PG2A) were measured by scanning densitometry after identification. One hundred and ninety-one patients were enrolled in the study. Specific IgE to all tomato extracts was evaluated by direct ELISA. Results Eighty (41.7%) individuals reported symptoms after contact/ingestion with tomato, most of them oral symptoms. Protein profile and allergen quantification of the 6 varieties showed differences in the allergen composition. Lyc e 3 was visualized in all extracts, being the most abundant in all cases. PG2A and OLP were identified in all peel extracts and in some pulp extracts. Higher specific IgE values were obtained for the Kumato variety. Immunoblot showed the existence of several bands with IgE-binding capacity, especially at 12 and 45-50 kDa. Conclusions The study demonstrates differences in the antigenic and allergenic profiles of the analyzed varieties and variations in their allergenic composition, affecting the diagnostic capacity. LTP and PG2A are important tomato allergens in our population but other IgE-binding capacity proteins are also involved.

Summary Background Tomatoes (Lycopersicon esculentum) are consumed world-wide. The prevalence of sensitization to tomatoes remains unknown. Objective To determine the prevalence of skin test reactivity to tomato and to describe the characteristics of tomato-sensitized subjects. Methods Individuals attending for the first time during the period of the study to six Allergy centres, located along the Mediterranean coast of Spain, reporting respiratory and/or cutaneous symptoms, were included. All patients were skin prick tested with a battery of inhalant allergens and with peel and pulp of Canary tomato extracts. Results The study included 1734 individuals (757 males, 977 females; 31.9±17.8 years old). The prevalence of sensitization to tomato was 6.52% (113 patients; 65 males, 48 females; 29.5±13 years old). The peel extract was positive in 110 patients and the pulp extract in 47 patients; three patients were positive exclusively to pulp. Only 1.8% of individuals reported symptoms with tomato; 44% of them had skin test negative to both extracts. Among tomato-sensitized subjects, 16% reported symptoms with tomato, 97% were sensitized to inhalant aeroallergens, including 84% to pollens (mainly Artemisia vulgaris and Platanus hybrida), with differences between Northern and Southern centres. Conclusions The prevalence found of skin test sensitivity to tomato is high. Peel extracts detected most of the sensitized subjects. Most of the sensitized subjects were asymptomatic and some patients reported symptoms without skin test sensitivity. Positive subjects were very frequently sensitized to pollens, suggesting allergen cross-reactivity. Regional differences may exist, possibly related to the pattern of sensitization to cross-reacting pollens.

Background Diagnosis of Alternaria alternata sensitization is hampered by the variability and complexity of fungal extracts, and thus simplification of the diagnostic procedures with purified allergens should be pursued. Objective We sought to compare A alternata extract and purified natural Alt a 1 (nAlt a 1) and recombinant Alt a 1 (rAlt a 1) allergens for their diagnostic value. Methods Forty-two patients allergic to A alternata , 10 atopic patients with negative skin prick test responses to A alternata extract, and 10 healthy subjects were investigated. Skin prick tests and determination of specific IgE levels were performed with nAlt a 1 and 2 different types of rAlt a 1: rbAlt a 1, expressed in Escherichia coli , and ryAlt a 1, expressed in the yeast Yarrowia lipolytica . Results Prevalence for Alt a 1, Alt a 2, and Alt a 11 by IgE dot-blot testing was 98%, 0%, and 15%, respectively, and therefore Alt a 1 was used as a marker for A alternata sensitization. Immunoblotting and inhibition analysis showed no IgE-binding differences between nAlt a 1 and rAlt a 1. The whole group of patients with allergy to A alternata had positive skin test reactions to purified allergens at 100 μg/mL, whereas no false-positive reactions were detected. Natural or ryAlt a 1 elicited a similar response in skin tests compared with A alternata extract, although a reduced reactivity was observed with rbAlt a 1. Specific IgE levels to nAlt a 1 or rAlt a 1 showed significant correlation and similar sensitivity and specificity. Conclusions Alt a 1, either in its natural or recombinant form, is sufficient for a reliable diagnosis of A alternata sensitization and induces skin prick reactivity comparable with that produced by A alternata extract.

Hypersensitivity to carmine (E120) has been identified as a cause of food intolerance and occupational asthma. We present a case of occupational asthma following exposure to carmine in a manufacturer of sausages and review the literature. Case Report: A 42-year-old non-atopic male presented with a 5-year history of rhinoconjunctivitis and asthma on occupational exposure to food additive dusts. Symptoms increased after work. The patient had been exposed for more than 20 years. Methods: Skin prick tests were performed with a battery of common inhalant allergens and spices. Cochineal, carmine lake and additive mixes used by the patient were extracted and subsequently used for skin prick test, bronchial provocation and in vitro measurements (specific IgE, Western blot and chromatographic fractionation). Results: Prick tests were positive to carmine and carmine-containing additives; carmine-specific IgE and bronchial challenge tests were also positive (PC20 = 0.0004 mg/ml and 1.6 kU/l). Western blot showed IgE binding to bands of about 30 kDa on cochineal extract and a diffuse pattern at 40–97 kDa on carmine. This result was confirmed by gel filtration chromatography and dot blot. Carmine completely inhibited IgE binding to cochineal extract. Discussion: Carmine is a potential sensitizer in an occupational setting: 18 cases of occupational asthma have been described to date. Carmine allergens are poorly defined; in general, proteins from cochineal not removed by the extraction process are considered as the main allergens in carmine. Our results are consistent with this, but show that these proteins may be subject to chemical modification.

Rationale Cypress spp. pollen induced pollinosis is increasing worldwide. The objective of this study was to characterize 4 Cupressus arizonica extracts using different solvents and in vivo and in vitro techniques. Methods Commercially available Cupressus arizonica pollen was used to prepare 4 types of extracts extracts. Extract A was extracted in PBS (0.01M); extract B in ammonium bicarbonate (40 mM); extract C, in ammonium bicarbonate (40 mM), followed by precipitation with ammonium sulfate; and extract D consisted of the supernatant obtained after the centrifugation of extract C. All extracts were characterized by SDS-PAGE and immunoblot. Cup a 1 content was measured by scanning densitometry. Thirty individuals were skin tested with the 4 extracts at 2 mg/ml of freeze-dried material. Specific IgE was evaluated by direct ELISA and immunoblots. Polysaccharide content was evaluated by the Anthrone method. Results The diagnosis capacity of extract C was better than of extracts A, B and D. All skin prick tests were negative with extract D. Extract C had the highest protein and major allergen content and induced greater wheal sizes than the other extracts (p Conclusions The increase in the protein and major allergen content and the removal of irrelevant material produces a better extract for the in vivo and in vitro diagnosis of C. arizonica induced respiratory allergies.

Rationale The Cucurbitaceae family includes melons, cucumbers, pumpkins and squash along with utilitarian, decorative and edible gourds. Minute soft hairs are present on the surface of the rind. Before melons are shipped to the market, they are brushed, cleaned and waxed to remove dirt and hairs. The objective was to investigate the prevalence of sensitization to melon (rind, pulp and hairs) in occupationally exposed workers. Methods A questionnaire, including a history of sensitization to aeroallergens and fruits and time of exposure, was administered to 47 workers (38 females, 9 males; mean age 32 years). All individuals were skin tested with extracts of melon hairs, 20 mg/ml, and rind and pulp at 10 mg/ml. Results Mean time of exposure was 1.6 years, although they were only directly exposed during the 3 month period of the harvest. The rest of the year they worked with other vegetables, including peaches, green peppers and tomatoes. A total of 6 individuals had positive skin tests to melon extracts (12.7%); 2 of these individuals had a positive skin test with the rind extract and a clinical history of oral allergy syndrome (4.3%); 4 individuals (8.5%) had a positive skin test with melon hair extract and had a clinical history of contact dermatitis. Exposure time in these individuals ranged from 5 to 8 years. Conclusions We have detected a prevalence of allergic contact dermatitis and sensitization to melon hairs of 8.5% among occupationally exposed individuals. Workers exposed to melons may become sensitized to melon allergens.

Rationale The objective of this study was to perform nasal provocation tests with standardized extracts of Dermatophagoides pteronyssinus , D. farinae , Lepidoglyphus destructor and Tyrophagus putrescentiae in a group of patients sensitized to Dermatophagoides spp. Methods A total of 25 patients with allergic rhinitis with or without asthma were entered in the study. All patients had positive skin tests to Dermatophagoides spp. Three groups of patients were studied. Group A consisted of 10 patients, who underwent nasal provocation tests with standardized extracts of D. pteronyssinus and D. farinae (100 HEP/ml). Group B consisted of 10 patients, 9 of whom were skin prick test positive to L. destructor . All 10 patients underwent a nasal provocation test with L. destructor and 5 with D. pteronyssinus . Group C, consisted of 5 patients, 2 of whom were skin test positive to T. putrescentiae . All individuals underwent nasal provocation tests with T. putrescentiae and Dermatophagoides spp. Specific IgE was measured by the CAP system. Results Group A: 7 patients had a positive provocation test with D. pteronyssinus and 8 with D. farinae . Group B: 6 patients had a positive provocation test with L. destructor . Group C: 3 patients had a positive provocation test with T. putrescentiae . Conclusions We confirm a high prevalence of positive nasal challenges to L. destructor and T. putrescentiae in patients sensitized to these mites and to Dermatophagoides spp. The clinical relevance of these mites species is suggested. In general, there was a good correlation between specific IgE levels and positive provocation tests.

El Greco y su interesante técnica pictórica me llevó a visitar en el verano de 1987 la Academia de Venecia. El objetivo de este estudio consistía en analizar la relación que este pintor había mantenido desde el punto de vista técnico con los pintores que trabajaron en la ciudad en la primera mitad del Siglo XV. Recorrer las salas de la Academia con una intención tan concreta que me hacía observar las obras con extrema atención permitió que me enfrentara con verdadero asombro a las grandes telas, obras de los Bellini, Mansueti, Carpaccio y otros artistas que protagonizaron la actividad artística veneciana del siglo XV, junto a unas pinturas de factura bien distinta realizadas a partir de mediados del mismo siglo por Giorgone, Tiziano, Tintoretto y otros pintores del momento. La magnitud de los cambios que podía apreciar entre unas obras y otras me llevó casi irremediablemente al trabajo de investigación que a continuación se expone. La búsqueda de factores técnicos que se relacionaron con un cambio plástico tan marcado desvelaba de alguna forma la incidencia que el soporte, en este caso la tela, había tenido en la expresión plástica de estos artistas. La nueva concepción de la obra pictórica que se estaba gestando se relacionaba con un notable cambio técnico que no se justifica únicamente por la utilización de la pintura al óleo que sustituye poco a poco a los temples como aglutinantes de sus pinturas. La repercusión de los tejidos empleados como soportes en la superficie plástica era evidente. De esta forma llegamos a concebir lo que sería el objetivo general del presente trabajo, el análisis de las posibilidades técnicas y expresivas de la tela como soporte de la pintura.

Altres ajuts: Robert Montal is supported by a FSEOM-Boehringer Ingelheim Grant. Carmen Andreu-Oller has received financial support through the "la Caixa" INPhINIT Fellowship Grant for Doctoral studies at Spanish Research Centres of Excellence, from "la Caixa" Banking Foundation (ID 100010434), fellowship code (LCF/BQ/IN17/ 11620024). Carla Montironi is a recipient of Josep Font grant from Hospital Clinic de Barcelona. Augusto Villanueva is supported by U.S. Department of Defense (CA150272P3) and Tisch Cancer Institute (Cancer Center Grant P30-CA196521). Manel Esteller is supported by CIBER 2016 CB16/12/00312 (CIBERONC); the Cellex Foundation; "la Caixa" Banking Foundation (LCF/PR/PR15/11100003). Josep M. Llovet is supported by National Cancer Institute (P30-CA196521). ; Generalitat de Catalunya, Departament de Salut, Projecte PERIS SLT-002-16-00374 The clinical utility of serum alpha-fetoprotein (AFP) in patients with hepatocellular carcinoma (HCC) is widely recognised. However, a clear understanding of the mechanisms of AFP overexpression and the molecular traits of patients with AFP-high tumours are not known. We assessed transcriptome data, whole-exome sequencing data and DNA methylome profiling of 520 HCC patients from two independent cohorts to identify distinct molecular traits of patients with AFP-high tumours (serum concentration > 400 ng/ml), which represents an accepted prognostic cut-off and a predictor of response to ramucirumab. Those AFP-high tumours (18% of resected cases) were characterised by significantly lower AFP promoter methylation (p < 0.001), significant enrichment of progenitor-cell features (CK19, EPCAM), higher incidence of BAP1 oncogene mutations (8.5% vs 1.6%) and lower mutational rates of CTNNB1 (14% vs 30%). Specifically, AFP-high tumours displayed significant activation of VEGF signalling (p < 0.001), which might provide the rationale for the reported benefit of ramucirumab in this subgroup of patients.