ILK mediates LPS-induced vascular adhesion receptor expression and subsequent leucocyte trans-endothelial migration†

Aims The inflammatory response to injurious agents is tightly regulated to avoid adverse consequences of inappropriate leucocyte accumulation or failed resolution. Lipopolysaccharide (LPS)-activated endothelium recruits leucocytes to the inflamed tissue through controlled expression of membrane-associated adhesion molecules. LPS responses in macrophages are known to be regulated by integrin-linked kinase (ILK); in this study, we investigated the role of ILK in the regulation of the LPS-elicited inflammatory response in endothelium. Methods and results This study was performed on immortalized mouse endothelial cells (EC) isolated from lung and coronary vasculature. Cells were thoroughly characterized and the role of ILK in the regulation of the LPS response was investigated by suppressing ILK expression using siRNA and shRNA technologies. Phenotypic and functional analyses confirmed that the immortalized cells behaved as true EC. LPS induced the expression of the inflammatory genes E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). ILK knockdown impaired LPS-mediated endothelial activation by preventing the induction of ICAM-1 and VCAM-1. Blockade of the LPS-induced response inhibited the inflammatory-related processes of firm adhesion and trans-endothelial migration of leucocytes. Conclusion ILK is involved in the expression of cell adhesion molecules by EC activated with the inflammatory stimulus LPS. This reduced expression modulates leucocyte adhesion to the endothelium and the extravasation process. This finding suggests ILK as a potential anti-inflammatory target for the development of vascular-specific treatments for inflammation-related disease

IL-10 released by a new inflammation-regulated lentiviral system efficiently attenuates zymosan-induced arthritis

We thank Dr Filip Lim for critical reading of the manuscript, and Dr S. Bartlett for English editing and helpful discussions. We also thank Drs. David Sancho and M. A. del Pozo for providing us with DCs and immortalized MEFs, respectively. AR is supported by Plan Nacional de Investigación Científica, Desarrollo e Innovación Tecnológica (I+D+I) and Instituto de Salud Carlos III (FIS; PI060122), the Spanish Ministry of Science and Innovation (MICINN;SAF2009-10691) and the Comunidad Autónoma de Madrid (S2006/BIO-0236 and S2010/BMD-2312). JMR is supported by MICINN (RECAVA RD06/0014/005) and by from Fundació La Marató de TV3 (Grant 080731

Mice Lacking Thyroid Hormone Receptor β Show Enhanced Apoptosis and Delayed Liver Commitment for Proliferation after Partial Hepatectomy

This is an open-access article distributed under the terms of the Creative Commons Attribution License.[Background]: The role of thyroid hormones and their receptors (TR) during liver regeneration after partial hepatectomy (PH) was studied using genetic and pharmacologic approaches. Roles in liver regeneration have been suggested for T3, but there is no clear evidence distinguishing the contribution of increased amounts of T3 from the modulation by unoccupied TRs. [Methodology/Principal Findings]: Mice lacking TR alpha 1/TR beta or TR beta alone fully regenerated liver mass after PH, but showed delayed commitment to the initial round of hepatocyte proliferation and transient but intense apoptosis at 48h post-PH, affecting similar to 30% of the remaining hepatocytes. Pharmacologically induced hypothyroidism yielded similar results. Loss of TR activity was associated with enhanced nitrosative stress in the liver remnant, due to an increase in the activity of the nitric oxide synthase (NOS) 2 and 3, caused by a transient decrease in the concentration of asymmetric dimethylarginine (ADMA), a potent NOS inhibitor. This decrease in the ADMA levels was due to the presence of a higher activity of dimethylarginineaminohydrolase-1 (DDAH-1) in the regenerating liver of animals lacking TR alpha 1/TR beta or TR beta. DDAH-1 expression and activity was paralleled by the activity of FXR, a transcription factor involved in liver regeneration and up-regulated in the absence of TR. [Conclusions/Significance]: We report that TRs are not required for liver regeneration; however, hypothyroid mice and TR beta-or TR alpha 1/TR beta-deficient mice exhibit a delay in the restoration of liver mass, suggesting a specific role for TRb in liver regeneration. Altered regenerative responses are related with a delay in the expression of cyclins D1 and E, and the occurrence of liver apoptosis in the absence of activated TRb that can be prevented by administration of NOS inhibitors. Taken together, these results indicate that TRb contributes significantly to the rapid initial round of hepatocyte proliferation following PH, and improves the survival of the regenerating liver at later times.This work was supported by grants BFU2008-02161, BFU2007-62402, SAF2007-60511, and SAF2007-60551 from MICINN; S-BIO-0283/2006 from Comunidad de Madrid and FIS-RECAVA RD06/0014/0025 to L.B.; and PI05.0050, PI080070, and the Fundacion Mutua Madrileña to S.H. RECAVA and Ciberehd are funded by the Instituto de Salud Carlos III. R. L-F. is supported by a fellowship from Instituto de Salud Carlos III. The CNIC is supported by the Spanish Ministry of Science and Innovation and the Pro-CNIC Foundation.Peer reviewe

The tumor suppressor ARF regulates innate immune responses in mice

The innate immune system is the first line of defense against invading organisms, and TLRs are the main sensors of microbial components, initiating signaling pathways that induce the production of proinflammatory cytokines and type I IFNs. An antiviral action for the tumor suppressor alternative reading frame (ARF) has been reported; however, the precise role of ARF in innate immunity is unknown. In this study, we show that ARF plays an important role in regulation of inflammatory responses. In peritoneal macrophages and bone marrow-derived macrophages from ARF-deficient animals, the induction of proinflammatory cytokines and chemokines by TLR ligands was severely impaired. The altered responses of ARF-/- cells to TLR ligands result from aberrant activation of intracellular signaling molecules including MAPKs, IkBa degradation, and NF-κB activation. Additionally, animals lacking ARF were resistant to LPS-induced endotoxic shock. This impaired activation of inflammation in ARF-/- mice was not restricted to TLRs, as it was also shown in response to non-TLR signaling pathways. Thus, ARF-/- mice were also unable to trigger a proper inflammatory response in experimental peritonitis or in 12-O-tetradecanoylphorbol-13-acetate-induced edema. Overexpression of ARF, but not its downstream target p53, rescued the ARF-deficient phenotype, increasing TLR4 levels and restoring inflammatory reaction. An increase in the E2F1 protein levels observed in ARF-/- macrophages at basal condition and after LPS stimulation may be involved in the impaired response in this system, as E2F1 has been described as an inflammatory suppressor. These results indicate that tumor suppressor ARF is a new regulator of inflammatory cell signaling. Copyright © 2011 by The American Association of Immunologists, Inc.This work was supported by Grant PI08.0070 from the Fondo de Investigaciones Sanitarias, MPY 1410/09 from Instituto de Salud Carlos III, and a Fundación Mutua Madrileña award (to S.H.). P.G.T. was supported by a Red Temática de Investigación Cooperativa en Enfermedades Cardiovasculares grant. A.L. was supported by Ramón y Cajal Fellowship RYC-2004-000367. Centro Nacional de Investigaciones Cardiovasculares is supported by the Spanish Ministry of Science and Innovation and the Pro-CNIC Foundation.Peer Reviewe

X Jornadas Internacionales de Innovación Universitaria, Villaviciosa de Odón, 11-12 de julio, 2013

SIN FINANCIACIÓNNo data 201

ILK mediates LPS-induced vascular adhesion receptor expression and subsequent leucocyte trans-endothelial migration

[Aims]: The inflammatory response to injurious agents is tightly regulated to avoid adverse consequences of inappropriate leucocyte accumulation or failed resolution. Lipopolysaccharide (LPS)-activated endothelium recruits leucocytes to the inflamed tissue through controlled expression of membrane-associated adhesion molecules. LPS responses in macrophages are known to be regulated by integrin-linked kinase (ILK); in this study, we investigated the role of ILK in the regulation of the LPS-elicited inflammatory response in endothelium. [Methods and results]: This study was performed on immortalized mouse endothelial cells (EC) isolated from lung and coronary vasculature. Cells were thoroughly characterized and the role of ILK in the regulation of the LPS response was investigated by suppressing ILK expression using siRNA and shRNA technologies. Phenotypic and functional analyses confirmed that the immortalized cells behaved as true EC. LPS induced the expression of the inflammatory genes E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). ILK knockdown impaired LPS-mediated endothelial activation by preventing the induction of ICAM-1 and VCAM-1. Blockade of the LPS-induced response inhibited the inflammatory-related processes of firm adhesion and trans-endothelial migration of leucocytes. [Conclusion]: ILK is involved in the expression of cell adhesion molecules by EC activated with the inflammatory stimulus LPS. This reduced expression modulates leucocyte adhesion to the endothelium and the extravasation process. This finding suggests ILK as a potential anti-inflammatory target for the development of vascular-specific treatments for inflammation-related diseases. © The Author 2009. For permissions please.This work was supported by Ministerio de Ciencia e Innovación from Spain, RYC-2004-000367 to A.L.; BFU2009-02161 and RECAVA to L.B.; PI080070 and the Fundación Mutua Madrileña to S.H.Peer Reviewe

Differential sensitivity to apoptosis among the cells that contribute to the atherosclerotic disease

Apoptosis plays an important role in a great number of pathological processes, including atherosclerotic disease. Although apoptosis occurs in the major cell types found in atherosclerotic lesions (e.g. macrophages, endothelial cells, and smooth muscle cells), the mechanism involved in this process differs depending on the stage, the localization and the cell composition of the plaque. In this study, we have compared the effects of different apoptotic inducers on the cells that form the atherosclerotic plaque. We have demonstrated that monocytes and macrophages are more susceptible to apoptosis than smooth muscle cells and endothelial cells. These findings provide insights about the potential role of apoptosis in the atherosclerotic disease and suggest strategies to treat vascular diseases by exploiting the differential sensitivity of cells to cell death. © 2007 Elsevier Inc. All rights reserved.M.Z. is supported by a Marie Curie OIF, P.G.T. is supported by a RECAVA and R.L.-F. by a FPU fellowship from the Spanish Ministry of Education and Science. S.H. is a FIS program investigator and is supported by Plan Nacional de Investigación Científica, Desarrollo e Innovación tecnológica (I+D+I) and Instituto de Salud Carlos III. This work was supported by Grants FIS (2002/3022) and PI05.0050 (2005) from FIS, and Fundación Mutua Madrileña to S.H. CNIC is supported by the Spanish Ministry of Health and Consumer Affairs and the Pro-CNIC Foundation.Peer Reviewe

Medios audiovisuales en la docencia actual

SIN FINANCIACIÓNNo data 201

Compuestos antitumorales

[EN] The invention provides a series of compounds with labdane-type diterpenoid structure that are capable of activating caspase-8 and therefore induce tumour cell apoptosis. The invention also relates to the use of said compounds for the treatment of diseases associated with an undesired cell proliferation and also to methods for inducing apoptosis in cells in culture and for identifying compounds with anti-apoptotic activity.[ES] La invención proporciona una serie de compuestos con estructura de diterpenoides de tipo labdano que son capaces de provocar la activación de caspasa-8 y, por tanto, inducen la apoptosis de células tumorales. La invención se relaciona también con el uso de dicho compuestos para el tratamiento de enfermedades asociadas a una proliferación celular indeseada así como a métodos para inducir la apoptosis en células en cultivo y para la identificación de compuestos con actividad anti-apoptótica.Peer reviewedFundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Consejo Superior de Investigaciones Científicas (España), Universidad Complutense de Madrid (UCM)A1 Solicitud de patentes con informe sobre el estado de la técnic