SUMOylation inhibition mediated by disruption of SUMO E1-E2 interactions confers plant susceptibility to necrotrophic fungal pathogens

Protein modification by SUMO modulates essential biological processes in eukaryotes. SUMOylation is facilitated by sequential action of the E1-activating, E2-conjugating, and E3-ligase enzymes. In plants, SUMO regulates plant development and stress responses, which are key determinants in agricultural productivity. To generate additional tools for advancing our knowledge about the SUMO biology, we have developed a strategy for inhibiting in vivo SUMO conjugation based on disruption of SUMO E1-E2 interactions through expression of E1 SAE2UFDCt domain. Targeted mutagenesis and phylogenetic analyses revealed that this inhibition involves a short motif in SAE2UFDCt highly divergent across kingdoms. Transgenic plants expressing the SAE2UFDCt domain displayed dose-dependent inhibition of SUMO conjugation, and have revealed the existence of a post-transcriptional mechanism that regulates SUMO E2 conjugating enzyme levels. Interestingly, these transgenic plants displayed increased susceptibility to necrotrophic fungal infections by Botrytis cinerea and Plectosphaerella cucumerina. Early after fungal inoculation, host SUMO conjugation was post-transcriptionally downregulated, suggesting that targeting SUMOylation machinery could constitute a novel mechanism for fungal pathogenicity. These findings support the role of SUMOylation as a mechanism involved in plant protection from environmental stresses. In addition, the strategy for inhibiting SUMO conjugation in vivo described in this study might be applicable in important crop plants and other non-plant organisms regardless of their genetic complexity

Thrombectomy within 8 hours after symptom onset in ischemic stroke

BACKGROUND: We aimed to assess the safety and efficacy of thrombectomy for the treatment of stroke in a trial embedded within a population-based stroke reperfusion registry. METHODS: During a 2-year period at four centers in Catalonia, Spain, we randomly assigned 206 patients who could be treated within 8 hours after the onset of symptoms of acute ischemic stroke to receive either medical therapy (including intravenous alteplase when eligible) and endovascular therapy with the Solitaire stent retriever (thrombectomy group) or medical therapy alone (control group). All patients had confirmed proximal anterior circulation occlusion and the absence of a large infarct on neuroimaging. In all study patients, the use of alteplase either did not achieve revascularization or was contraindicated. The primary outcome was the severity of global disability at 90 days, as measured on the modified Rankin scale (ranging from 0 [no symptoms] to 6 [death]). Although the maximum planned sample size was 690, enrollment was halted early because of loss of equipoise after positive results for thrombectomy were reported from other similar trials. RESULTS Thrombectomy reduced the severity of disability over the range of the modified Rankin scale (adjusted odds ratio for improvement of 1 point, 1.7; 95% confidence interval [CI], 1.05 to 2.8) and led to higher rates of functional independence (a score of 0 to 2) at 90 days (43.7% vs. 28.2%; adjusted odds ratio, 2.1; 95% CI, 1.1 to 4.0). At 90 days, the rates of symptomatic intracranial hemorrhage were 1.9% in both the thrombectomy group and the control group (P = 1.00), and rates of death were 18.4% and 15.5%, respectively (P = 0.60). Registry data indicated that only eight patients who met the eligibility criteria were treated outside the trial at participating hospitals. CONCLUSIONS: Among patients with anterior circulation stroke who could be treated within 8 hours after symptom onset, stent retriever thrombectomy reduced the severity of post-stroke disability and increased the rate of functional independence

The avoidance of G-CSF and the addition of prophylactic corticosteroids after autologous stem cell transplantation for multiple myeloma patients appeal for the at-home setting to reduce readmission for neutropenic fever

Autologous stem cell transplantation (ASCT) remains the standard of care for young multiple myeloma (MM) patients; indeed, at-home ASCT has been positioned as an appropriate therapeutic strategy. However, despite the use of prophylactic antibiotics, neutropenic fever (NF) and hospital readmissions continue to pose as the most important limitations in the outpatient setting. It is possible that the febrile episodes may have a non-infectious etiology, and engraftment syndrome could play a more significant role. The aim of this study was to analyze the impact of both G-CSF withdrawal and the addition of primary prophylaxis with corticosteroids after ASCT. Between January 2002 and August 2018, 111 MM patients conditioned with melphalan were managed at-home beginning +1 day after ASCT. Three groups were established: Group A (n = 33) received standard G-CSF post-ASCT; group B (n = 32) avoided G-CSF post-ASCT; group C (n = 46) avoided G-CSF yet added corticosteroid prophylaxis post-ASCT. The incidence of NF among the groups was reduced (64%, 44%, and 24%; P2 (OR 6.1; P = 0.002) and G-CSF avoidance plus corticosteroids (OR 0.1; P<0.001); and for hospital readmission: age �60 years (OR 14.6; P = 0.04) and G-CSF avoidance plus corticosteroids (OR 0.07; P = 0.05. G-CSF avoidance and corticosteroid prophylaxis post ASCT minimize the incidence of NF in MM patients undergoing at-home ASCT. This approach should be explored in a prospective randomized clinical trial

Population-based multicase-control study in common tumors in Spain (MCC-Spain): rationale and study design

Introduction: We present the protocol of a large population-based case-control study of 5 common tumors in Spain (MCC-Spain) that evaluates environmental exposures and genetic factors. Methods: Between 2008-2013, 10,183 persons aged 20-85 years were enrolled in 23 hospitals and primary care centres in 12 Spanish provinces including 1,115 cases of a new diagnosis of prostate cancer, 1,750 of breast cancer, 2,171 of colorectal cancer, 492 of gastro-oesophageal cancer, 554 cases of chronic lymphocytic leukaemia (CLL) and 4,101 population-based controls matched by frequency to cases by age, sex and region of residence. Participation rates ranged from 57% (stomach cancer) to 87% (CLL cases) and from 30% to 77% in controls. Participants completed a face-to-face computerized interview on sociodemographic factors, environmental exposures, occupation, medication, lifestyle, and personal and family medical history. In addition, participants completed a self-administered food-frequency questionnaire and telephone interviews. Blood samples were collected from 76% of participants while saliva samples were collected in CLL cases and participants refusing blood extractions. Clinical information was recorded for cases and paraffin blocks and/or fresh tumor samples are available in most collaborating hospitals. Genotyping was done through an exome array enriched with genetic markers in specific pathways. Multiple analyses are planned to assess the association of environmental, personal and genetic risk factors for each tumor and to identify pleiotropic effects. Discussion: This study, conducted within the Spanish Consortium for Biomedical Research in Epidemiology & Public Health (CIBERESP), is a unique initiative to evaluate etiological factors for common cancers and will promote cancer research and prevention in Spain.The study was partially funded by the “Accion Transversal del Cancer”, approved on the Spanish Ministry Council on the 11th October 2007, by the Instituto de Salud Carlos III-FEDER (PI08/1770, PI08/0533, PI08/1359, PS09/00773, PS09/01286, PS09/01903, PS09/02078, PS09/01662, PI11/01403, PI11/01889, PI11/00226, PI11/01810, PI11/02213, PI12/00488, PI12/00265, PI12/01270, PI12/00715, PI12/00150), by the Fundación Marqués de Valdecilla (API 10/09), by the ICGC International Cancer Genome Consortium CLL, by the Junta de Castilla y León (LE22A10-2), by the Consejería de Salud of the Junta de Andalucía (PI-0571), by the Conselleria de Sanitat of the Generalitat Valenciana (AP 061/10), by the Recercaixa (2010ACUP 00310), by the Regional Government of the Basque Country by European Commission grants FOOD-CT- 2006-036224-HIWATE, by the Spanish Association Against Cancer (AECC) Scientific Foundation, by the The Catalan Government DURSI grant 2009SGR1489

Common variants in Alzheimer’s disease and risk stratification by polygenic risk scores

Funder: Funder: Fundación bancaria ‘La Caixa’ Number: LCF/PR/PR16/51110003 Funder: Grifols SA Number: LCF/PR/PR16/51110003 Funder: European Union/EFPIA Innovative Medicines Initiative Joint Number: 115975 Funder: JPco-fuND FP-829-029 Number: 733051061Genetic discoveries of Alzheimer's disease are the drivers of our understanding, and together with polygenetic risk stratification can contribute towards planning of feasible and efficient preventive and curative clinical trials. We first perform a large genetic association study by merging all available case-control datasets and by-proxy study results (discovery n = 409,435 and validation size n = 58,190). Here, we add six variants associated with Alzheimer's disease risk (near APP, CHRNE, PRKD3/NDUFAF7, PLCG2 and two exonic variants in the SHARPIN gene). Assessment of the polygenic risk score and stratifying by APOE reveal a 4 to 5.5 years difference in median age at onset of Alzheimer's disease patients in APOE ɛ4 carriers. Because of this study, the underlying mechanisms of APP can be studied to refine the amyloid cascade and the polygenic risk score provides a tool to select individuals at high risk of Alzheimer's disease

TFG 2013/2014

Amb aquesta publicació, EINA, Centre universitari de Disseny i Art adscrit a la Universitat Autònoma de Barcelona, dóna a conèixer el recull dels Treballs de Fi de Grau presentats durant el curs 2013-2014. Voldríem que un recull com aquest donés una idea més precisa de la tasca que es realitza a EINA per tal de formar nous dissenyadors amb capacitat de respondre professionalment i intel·lectualment a les necessitats i exigències de la nostra societat. El treball formatiu s’orienta a oferir resultats que responguin tant a paràmetres de rigor acadèmic i capacitat d’anàlisi del context com a l’experimentació i la creació de nous llenguatges, tot fomentant el potencial innovador del disseny.Con esta publicación, EINA, Centro universitario de diseño y arte adscrito a la Universidad Autónoma de Barcelona, da a conocer la recopilación de los Trabajos de Fin de Grado presentados durante el curso 2013-2014. Querríamos que una recopilación como ésta diera una idea más precisa del trabajo que se realiza en EINA para formar nuevos diseñadores con capacidad de responder profesional e intelectualmente a las necesidades y exigencias de nuestra sociedad. El trabajo formativo se orienta a ofrecer resultados que respondan tanto a parámetros de rigor académico y capacidad de análisis, como a la experimentación y la creación de nuevos lenguajes, al tiempo que se fomenta el potencial innovador del diseño.With this publication, EINA, University School of Design and Art, affiliated to the Autonomous University of Barcelona, brings to the public eye the Final Degree Projects presented during the 2013-2014 academic year. Our hope is that this volume might offer a more precise idea of the task performed by EINA in training new designers, able to speak both professionally and intellectually to the needs and demands of our society. The educational task is oriented towards results that might respond to the parameters of academic rigour and the capacity for contextual analysis, as well as to considerations of experimentation and the creation of new languages, all the while reinforcing design’s innovative potential

Multiancestry analysis of the HLA locus in Alzheimer’s and Parkinson’s diseases uncovers a shared adaptive immune response mediated by HLA-DRB1*04 subtypes

Across multiancestry groups, we analyzed Human Leukocyte Antigen (HLA) associations in over 176,000 individuals with Parkinson’s disease (PD) and Alzheimer’s disease (AD) versus controls. We demonstrate that the two diseases share the same protective association at the HLA locus. HLA-specific fine-mapping showed that hierarchical protective effects of HLA-DRB1*04 subtypes best accounted for the association, strongest with HLA-DRB1*04:04 and HLA-DRB1*04:07, and intermediary with HLA-DRB1*04:01 and HLA-DRB1*04:03. The same signal was associated with decreased neurofibrillary tangles in postmortem brains and was associated with reduced tau levels in cerebrospinal fluid and to a lower extent with increased Aβ42. Protective HLA-DRB1*04 subtypes strongly bound the aggregation-prone tau PHF6 sequence, however only when acetylated at a lysine (K311), a common posttranslational modification central to tau aggregation. An HLA-DRB1*04-mediated adaptive immune response decreases PD and AD risks, potentially by acting against tau, offering the possibility of therapeutic avenues

Caracterización molecular y funcional de Small-Ubiquitin-related MOdifier en Arabidopsis thaliana

[spa] SUMO, Small Ubiquitin-related MOdifier, es un modificador post-traduccional de la familia de la Ubiquitina, ya que presentan estructuras y mecanismo de conjugación conservados. La unión de SUMO al sustrato puede modificar la actividad, localización y estabilidad de la proteína diana. En plantas, la conjugación de SUMO es un proceso esencial durante los primeros estadíos del desarrollo embrionario y que regula las respuestas de las plantas a hormonas y su adaptación frente a diferentes estreses abióticos y bióticos. La conjugación de SUMO al sustrato es el resultado de la acción secuencial de tres reacciones enzimáticas: la activación de SUMO, catalizada por el heterodímero SAE2/SAE1, la conjugación a la enzima conjugadora SCE1 y su transferencia al sustrato en una reacción facilitada por E3 ligasas. En Arabidopsis, existen cuatro isoformas de SUMO, SUMO1, 2, 3 y 4, y dos isoformas de la subunidad pequeña de la enzima activadora, SAE1a y b. Con el objetivo de analizar las bases moleculares de la función de SUMO in vivo, se ha procedido a la caracterización molecular de la maquinaria de SUMOylación. El análisis bioquímico de los parálogos de SUMO de Arabidopsis ha puesto de manifiesto la existencia de un mecanismo de regulación que permite la conjugación preferencial de las isoformas esenciales AtSUMO1/2. Este mecanismo consiste en una mayor afinidad de la enzima activadora E1 por SUMO1/2, de manera que la primera enzima de la vía seleccionaría la isoforma de SUMO que entra en la cascada de conjugación. Además, en Arabidopsis, existen dos isoformas de la enzima activadora E1 que difieren en la composición de la subunidad pequeña, SAE1a y SAE1b. Plantas mutantes de Arabidopsis que carecen de la subunidad SAE1a presentan deficiencias en las respuestas frente a estrés térmico e hídrico, sugiriendo que la subunidad pequeña podría tener un papel regulador en el mantenimiento de la homeostasis de SUMO in vivo. En resumen, los resultados indican que la enzima E1 activadora regularía la conjugación de SUMO mediante la selección de las isoformas de SUMO que entran en la cascada de conjugación y la cinética de conjugación mediante la expresión diferencial de las isoformas SAE1a y SAE1b. Resultados adicionales indican la existencia de modificaciones post-traduccionales que podrían regular la actividad de la enzima activadora in vivo. Por un lado, se ha identificado un procesamiento del extremo C-terminal de la enzima activadora que participa en la regulación de la localización subcelular de esta enzima. Por otro lado, se ha identificado la SUMOilación de diferentes componentes de la maquinaria de SUMO in vitro, entre los que se encuentran SAE2, SAE1, SUMO y SCE1. Estas modificaciones podrían estar regulando la actividad, estabilidad y localización de estas enzimas in vivo. Los resultados obtenidos han sentado las bases para el desarrollo de una herramienta molecular que permite la inhibición moderada de la SUMOylación in vivo. Esta herramienta consiste en la generación de una forma de SAE2 que actúa como dominante negativo, bloqueando la transferencia de SUMO desde la enzima activadora a la conjugadora. La expresión de este dominante negativo mimetiza los defectos de desarrollo característicos de plantas mutantes que presentan una SUMOilación disminuida, validando esta estrategia para el estudio de la función de SUMO en plantas in vivo. Mediante el uso de estas plantas, se ha establecido que un sistema de SUMOilación funcional es necesario para la activación de las respuestas de defensa de las plantas frente a patógenos. Además, como abordaje adicional para estudiar la función biológica de SUMO, se han realizado aproximaciones proteómicas en semilla y se han identificado proteínas potenciales sustrato de SUMO que podrían regular transiciones de desarrollo en semilla.[eng] SUMO, Small Ubiquitin-related Modifier, is a post-translational modifier belonging to the ubiquitin family, as they present preserved structures and mechanism of conjugation. In plants, SUMO conjugation is essential in the early stages of embryonic development and regulates plant responses to hormones and abiotic and biotic stresses. SUMO conjugation is the result of the three sequential enzymatic reactions: SUMO activation catalyzed heterodimer SAE2 / SAE1, conjugation to SCE1 conjugating enzyme and its transfer to the substrate in a reaction facilitated by E3 ligases. In Arabidopsis, there are four SUMO isoforms, SUMO1, 2, 3 and 4, and two isoforms of the small subunit of the activating-enzyme, SAE1a and b. In order to analyze the molecular basis of the role of SUMO in vivo, we performed the molecular characterization of SUMOylation machinery. Biochemical analysis of the Arabidopsis SUMO paralogs revealed the existence of a regulatory mechanism that allows the preferential conjugation of the essential isoforms AtSUMO1 / 2, which is mediated by the E1 activating enzyme. Also, there are two isoforms of the activating enzyme E1 which differ in the composition of the small subunit, SAE1a and SAE1b. We have shown that the SAE1a subunit is necessary for maintaining SUMOylation homeostasis in vivo. In summary, the results indicate that the E1 regulates SUMO conjugation by selecting SUMO isoforms entering the conjugation pathway and the conjugation rate by the differential expression of SAE1a/b isoforms. Additional results indicate that post-translational modifications may regulate the activity of the E1-activating enzyme. We have identified a processing of the E1 C-terminus involved in the regulation of its subcellular localization. Furthermore, we have identified the SUMOylation of different components of SUMO machinery, including SAE2, SAE1 SUMO and SCE1. We have advanced our knowledge of the SUMO biology by using a molecular tool that allows moderate inhibition of in vivo SUMOylation. We have engineered an E1 dominant negative which mimics plant responses typical of SUMOylation mutants. By using these plants, we have uncovered a new role of SUMOylation in defense responses against pathogens. Furthermore, proteomic approaches have been identified potential SUMO protein substrates that could regulate seed developmental transitions

Kinetic Analysis of Plant SUMO Conjugation Machinery

Plant SUMO conjugation is an essential posttranslational modification involved in plant development and responses to environmental stress. Most likely, this biological diversification is supported by a functional specialization of the different isoforms of the SUMO conjugation machinery. For instance, the two essential Arabidopsis SUMO isoforms, SUMO1/2, display higher conjugation rate than SUMO3 and 5, which are not essential, linking their specific biochemical properties to their biological role. To study the biochemical properties of plant SUMO conjugation systems, quantitative biochemical assays must be performed. We will present a detailed protocol for reconstituting an in vitro SUMO conjugation assay covering all steps from protein preparation to assay development.This work was supported by the European Research Council (grant ERC-2007-StG-205927) and Departament d’Innovació, Universitats i Empresa from the Generalitat de Catalunya (Xarxa de Referència en Biotecnologia and 2014 SGR 447). L.C.M was supported by research contract through the CRAG. This article is based upon work from COST Action (PROTEOSTASIS BM1307), supported by COST (European Cooperation in Science and Technology).Peer reviewe