MECHANISM OF SPLICING REGULATION BY THE MEIOSIS ENHANCER FACTOR Mer1p IN YEAST Saccharomyces cerevisiae

In eukaryotes, genes are presented in a series of coding and non-coding DNA regions (exons/introns) that are transcribed into a premature RNA (pre-mRNA). Introns can be removed from the mature premRNA, before its translation into proteins, in a process called splicing. The splicing reaction occurs in two highly regulated transesterification reactions inside of the cell nucleus, and it is catalyzed by the Spliceosome, involving the binding and release of five small nuclear ribonucleoprotein particles (snRNPs). While some introns are constitutively spliced, others can be alternatively spliced, giving different exon combinations and therefore different proteins, increasing the protein diversity of the species. In humans, misregulation of alternative splicing can result in the production of aberrant proteins, some of which may produce cancer or other severe diseases. In yeast, alternative splicing is regulated by different splicing factors, such as Mer1p. Mer1p is expressed during meiosis in the yeast Saccharomyces cerevisiae and activates the splicing in at least three different genes (AMA1, MER2, and MER3), which contain a conserved intronic splicing enhancer sequence. Previous results have shown that Mer1p is able to interact with the pre-mRNA and with specific proteins of the U1 and U2 snRNPs. However, the specific molecular mechanisms by which Mer1p activates splicing remained unknown. The objective of this work is to determine how Mer1p regulates the splicing of its targets, and how different splicing factors modulate Mer1p activity. Using biochemistry and genetics, the data presented in this work indicate that Mer1p recruits the snRNPs U1, U2 and U6, to pre-mRNA. This recruitment of the snRNPs is dependent of the U1 snRNP protein Nam8p and the U2 snRNP protein Snu17p, but independent on the branchpoint region or ATP. Furthermore, Mer1p accelerates and stabilizes the formation of the early complexes of the spliceosome. Finally, U1 and U2 are recruited to the pre-mRNA at the same time, emerging a new alternative hypothesis of splicing regulation that can be applied to other enhancer regulators and that differs from the classical model of stepwise assembly of the snRNP

La estructura resistente en la arquitectura actual (continuación)

The article addresses the relationship between a building.s support structure and the building itself in the context of the latest architectural tendencies. Today.s civil engineering has developed strength capabilities in terms of materials, structural layouts and construction processes so highly that there is a structural solution for any architectural approach, no matter how novel . or preposterous. The outcome may be the disconnection between the spatial and the structural conceptions of a building. The more novel a proposal, the more intensely architect and engineer must work together . Structure is not just something that any building should have, such as air conditioning, but in the larger picture is unessential. Most of today.s architecture: minimalist, deconstructionist, or high-tech, is fundamentally structural. Polytechnic architects should be distinguished from fine arts formalizers, who ultimately formalize better and more freely. A number of recent examples of buildings are reviewed in this light, grouped under the following headings: de-construction; high-rises; polyhedrons, cubes and so forth; Shigeru Ban and orthodox theories of strength; and finally, soft forms.En el artículo se plantea la relación entre la estructura resistente de un edificio y el edificio mismo según las últimas tendencias arquitectónicas. Actualmente la ingeniería civil ha desarrollado de tal forma las posibilidades resistentes, en cuanto a materiales, disposiciones estructurales y procesos constructivos que cualquier planteamiento arquitectónico, por novedoso -o disparatado- que sea, tiene solución estructural. Esto nos puede llevar a que la concepción espacial del edificio se separe de la estructural. Arquitecto e ingeniero deben trabajar en común, más intensamente cuanto más novedosa sea la propuesta. La estructura no es algo que todo edificio debe tener, como el aire acondicionado, pero que no importa demasiado. Los caminos de la arquitectura actual: minimal, deconstrucción, high-tech son esencialmente estructurales. El arquitecto politécnico debe distinguirse de un formalizador de bellas artes, que acabará formalizando mejor y más libremente. A la luz de estas ideas se revisan múltiples ejemplos recientes de edificios agrupados según distintos apartados: deconstrucción; edificios en altura; poliedros, cubos, etc.; Shigeru Ban y la ortodoxia resistente; y, finalmente, formas blandas

Developmental characterization of the microRNA-specific C. elegans Argonautes alg-1 and alg-2.

The genes alg-1 and alg-2 (referred to as "alg-1/2") encode the Argonaute proteins affiliated to the microRNA (miRNA) pathway in C. elegans. Bound to miRNAs they form the effector complex that effects post-transcriptional gene silencing. In order to define biological features important to understand the mode of action of these Argonautes, we characterize aspects of these genes during development. We establish that alg-1/2 display an overlapping spatio-temporal expression profile and shared association to a miRNAs set, but with gene-specific predominant expression in various cells and increased relative association to defined miRNAs. Congruent with their spatio-temporal coincidence and regardless of alg-1/2 drastic post-embryonic differences, only loss of both genes leads to embryonic lethality. Embryos without zygotic alg-1/2 predominantly arrest during the morphogenetic process of elongation with defects in the epidermal-muscle attachment structures. Altogether our results highlight similarities and specificities of the alg-1/2 likely to be explained at different cellular and molecular levels

The target antigen determines the mechanism of acquired resistance to T cell-based therapies

Cancer; Antigen; ResistanceCáncer; Antígeno; ResistenciaCàncer; Antigen; ResistènciaDespite the revolution of immunotherapy in cancer treatment, patients eventually progress due to the emergence of resistance. In this scenario, the selection of the tumor antigen can be decisive in the success of the clinical response. T cell bispecific antibodies (TCBs) are engineered molecules that include binding sites to the T cell receptor and to a tumor antigen. Using gastric CEA+/HER2+ MKN45 cells and TCBs directed against CEA or HER2, we show that the mechanism of resistance to a TCB is dependent on the tumor antigen. Acquired resistant models to a high-affinity-CEA-targeted TCB exhibit a reduction of CEA levels due to transcriptional silencing, which is reversible upon 5-AZA treatment. In contrast, a HER2-TCB resistant model maintains HER2 levels and exhibit a disruption of the interferon-gamma signaling. These results will help in the design of combinatorial strategies to increase the efficacy of cancer immunotherapies and to anticipate and overcome resistances.This work was supported by Asociación Española Contra el Cancer (AECC), Breast Cancer Research Foundation (BCRF-21-008), and Instituto de Salud Carlos III (PI19/01181). A.M.S. was funded by the Spanish Government (PFIS FI20/00188). B.M. was funded by a fellowship from PERIS (Departament de Salut, Generalitat de Catalunya). M.R.A. was funded by Agency for Management of University and Research Grants (AGAUR, 2022 FI_B2 00080). P.O.R. was funded by the BBVA. E.J.A. was funded by the AECC (POSTD211413AREN). VHIO acknowledges the Cellex Foundation for providing research facilities and equipment, the Centro de Investigación Biomédica en Red de Cáncer (CIBERONC) from the Institute of Health Carlos III (ISCIII), and the Department of Health (Generalitat de Catalunya, SLT008/18/00198 SLT008/18/00205) for their support on this research. The authors acknowledge financial support from the State Agency for Research (Agencia Estatal de Investigación) (CEX2020-001024-S/AEI/10.13039/501100011033) and for the Cancer Immunology and Immunotherapy (CAIMI-2) program funded by BBVA Foundation. We would like to remark the funding from B.M PERIS (Spain). The authors thank Dr. Anne Freimoser-Grundschober and Roche for helping provide the TCBs. The graphical abstract was created with BioRender.com

Therapy-Induced Senescence Enhances the Efficacy of HER2-Targeted Antibody–Drug Conjugates in Breast Cancer

Eficàcia; Conjugats de fàrmacs; Càncer de mamaEficacia; Conjugados de medicamentos; Cáncer de mamaEfficacy; Drug conjugates; Breast cancerAntibody–drug conjugates (ADC) are antineoplastic agents recently introduced into the antitumor arsenal. T-DM1, a trastuzumab-based ADC that relies on lysosomal processing to release the payload, is approved for HER2-positive breast cancer. Next-generation ADCs targeting HER2, such as [vic-]trastuzumab duocarmazine (SYD985), bear linkers cleavable by lysosomal proteases and membrane-permeable drugs, mediating a bystander effect by which neighboring antigen-negative cells are eliminated. Many antitumor therapies, like DNA-damaging agents or CDK4/6 inhibitors, can induce senescence, a cellular state characterized by stable cell-cycle arrest. Another hallmark of cellular senescence is the enlargement of the lysosomal compartment. Given the relevance of the lysosome to the mechanism of action of ADCs, we hypothesized that therapies that induce senescence would potentiate the efficacy of HER2-targeting ADCs. Treatment with the DNA-damaging agent doxorubicin and CDK4/6 inhibitor induced lysosomal enlargement and senescence in several breast cancer cell lines. While senescence-inducing drugs did not increase the cytotoxic effect of ADCs on target cells, the bystander effect was enhanced when HER2-negative cells were cocultured with HER2-low cells. Knockdown experiments demonstrated the importance of cathepsin B in the enhanced bystander effect, suggesting that cathepsin B mediates linker cleavage. In breast cancer patient-derived xenografts, a combination treatment of CDK4/6 inhibitor and SYD985 showed improved antitumor effects over either treatment alone. These data support the strategy of combining next-generation ADCs targeting HER2 with senescence-inducing therapies for tumors with heterogenous and low HER2 expression. Significance: Combining ADCs against HER2-positive breast cancers with therapies that induce cellular senescence may improve their therapeutic efficacy by facilitating a bystander effect against antigen-negative tumor cells.This work was supported by Breast Cancer Research Foundation (BCRF-20-008), Instituto de Salud Carlos III (project reference numbers AC15/00062, CB16/12/00449 and PI19/01181), the EC under the framework of the ERA-NET TRANSCAN-2 initiative co-financed by FEDER, Fundación Mutua Madrileña and Asociación Española Contra el Cáncer. S. Duro-Sánchez is supported by the Spanish Ministerio de Universidades by the grant Formación de Profesorado Universitario (FPU20/05388). A. Esteve-Codina is funded by ISCIII /MINECO (PT17/0009/0019) and co-funded by FEDER. The authors acknowledge Alyson MacInnes for reviewing and editing the article

RICORS2040 : The need for collaborative research in chronic kidney disease

Chronic kidney disease (CKD) is a silent and poorly known killer. The current concept of CKD is relatively young and uptake by the public, physicians and health authorities is not widespread. Physicians still confuse CKD with chronic kidney insufficiency or failure. For the wider public and health authorities, CKD evokes kidney replacement therapy (KRT). In Spain, the prevalence of KRT is 0.13%. Thus health authorities may consider CKD a non-issue: very few persons eventually need KRT and, for those in whom kidneys fail, the problem is 'solved' by dialysis or kidney transplantation. However, KRT is the tip of the iceberg in the burden of CKD. The main burden of CKD is accelerated ageing and premature death. The cut-off points for kidney function and kidney damage indexes that define CKD also mark an increased risk for all-cause premature death. CKD is the most prevalent risk factor for lethal coronavirus disease 2019 (COVID-19) and the factor that most increases the risk of death in COVID-19, after old age. Men and women undergoing KRT still have an annual mortality that is 10- to 100-fold higher than similar-age peers, and life expectancy is shortened by ~40 years for young persons on dialysis and by 15 years for young persons with a functioning kidney graft. CKD is expected to become the fifth greatest global cause of death by 2040 and the second greatest cause of death in Spain before the end of the century, a time when one in four Spaniards will have CKD. However, by 2022, CKD will become the only top-15 global predicted cause of death that is not supported by a dedicated well-funded Centres for Biomedical Research (CIBER) network structure in Spain. Realizing the underestimation of the CKD burden of disease by health authorities, the Decade of the Kidney initiative for 2020-2030 was launched by the American Association of Kidney Patients and the European Kidney Health Alliance. Leading Spanish kidney researchers grouped in the kidney collaborative research network Red de Investigación Renal have now applied for the Redes de Investigación Cooperativa Orientadas a Resultados en Salud (RICORS) call for collaborative research in Spain with the support of the Spanish Society of Nephrology, Federación Nacional de Asociaciones para la Lucha Contra las Enfermedades del Riñón and ONT: RICORS2040 aims to prevent the dire predictions for the global 2040 burden of CKD from becoming true

La estructura resistente de los edificios altos

If we only think about the resistance problem, a building could be considered as a great vertical console. The resistant exigencies of this huge console are as pressing as the building grows up and its fitting to the functional needs are more difficult. This study reviews the different resistant typologies, from the normal porch structure of medium size buildings to the "tube into the tube" and total laminar structures corresponding to the higher buildings which are already built, or are to be built. It also makes an analysis on the manner in which each typology resist, and shows examples of constructed buildings.<br><br>Si consideramos únicamente el problema resistente, un edificio alto podría considerarse como una gran ménsula vertical. Las exigencias resistentes de esta gran ménsula son más apremiantes conforme el edificio crece en altura y su acoplamiento a las necesidades funcionales del edificio más difícil. En este artículo se pasa revista a las diversas tipologías resistentes, desde la estructura aporticada normal de los edificios de altura moderada hasta el "tubo en el tubo" y estructuras laminares totales correspondientes a los más altos edificios construidos y por construir. Se hace un análisis de la forma de resistir de cada una de las tipologías ilustrándolas con ejemplos de edificios construidos