Double-edged lipid nanoparticles combining liposome-bound TRAIL and encapsulated doxorubicin showing an extraordinary synergistic pro-apoptotic potential

Although TRAIL (TNF-related apoptosis-inducing ligand, also known as Apo2L) was described as capable of inducing apoptosis in transformed cells while sparing normal cells, limited results obtained in clinical trials has limited its use as an anti-tumor agent. Consequently, novel TRAIL formulations with enhanced bioactivity are necessary for overcoming resistance to conventional soluble TRAIL (sTRAIL) exhibited by many primary tumors. Our group has generated artificial liposomes with sTRAIL anchored on their surface (large unilamellar vesicle (LUV)-TRAIL), which have shown a greater cytotoxic activity both in vitro and in vivo when compared to sTRAIL against distinct hematologic and epithelial carcinoma cells. In this study, we have improved LUV-TRAIL by loading doxorubicin (DOX) in its liposomal lumen (LUVDOX-TRAIL) in order to improve their cytotoxic potential. LUVDOX-TRAIL killed not only to a higher extent, but also with a much faster kinetic than LUV-TRAIL. In addition, the concerted action of the liposomal DOX and TRAIL was specific of the liposomal DOX and was not observed when with soluble DOX. The cytotoxicity induced by LUVDOX-TRAIL was proven to rely on two processes due to different molecular mechanisms: a dynamin-mediated internalization of the doxorubicin-loaded particle, and the strong activation of caspase-8 exerted by the liposomal TRAIL. Finally, greater cytotoxic activity of LUVDOX-TRAIL was also observed in vivo in a tumor xenograft model. Therefore, we developed a novel double-edged nanoparticle combining the cytotoxic potential of DOX and TRAIL, showing an exceptional and remarkable synergistic effect between both agents

Uso de plataformas microfluídicas para la valoración de drogas anti-tumorales

La nueva formulación de TRAIL basada en su asociación a vesículas lipídicas de tipo LUV (Large Unilamellar Vesicles) denominada LUV-TRAIL ha aumentado la capacidad citotóxica de TRAIL frente a distintas células tumorales. En este trabajo se ha comparado la citotoxicidad de TRAIL soluble y LUV-TRAIL en líneas celulares tumorales de pulmón (A549) y de riñón (Caki-1). La citotoxicidad ha sido estudiada tanto mediante cultivos celulares convencionales como mediante cultivos tridimensionales empleando plataformas microfluídicas. Los resultados obtenidos en ambos tipos de cultivos celulares muestran que los LUV-TRAIL presentan una mayor capacidad citotóxica que TRAIL soluble validando esta nueva formulación de este ligando mortal y demostrando que el empleo de plataformas microfluídicas puede ser muy útil en el estudio de diferentes compuestos anti-tumorales ya que permiten crear in vitro modelos tumorales que se asemejan a las condiciones in vivo sin el uso de animales de experimentación

ESTUDIO DE LA UTILIDAD DE LA IL-6 COMO MARCADOR DE LA ACTIVIDAD EN LUPUS ERITEMATOSO SISTÉMICO

Introducción: El Lupus eritematoso sistémico (LES) es una enfermedad autoinmune que cursa con una clínica heterogénea y fluctuante. Actualmente se emplea el índice SLEDAI para medir objetivamente la actividad de la enfermedad, considerando marcadores analíticos de actividad el descenso del complemento y la presencia de Autoanticuerpos anti-dsDNA en suero. La interleuquina 6 (IL-6), es una citoquina proinflamatoria implicada en distintos aspectos de la fisiopatología del lupus, por lo que este estudio busca conocer si puede ser un biomarcador del lupus.Objetivos: El objetivo principal es analizar la utilidad de la IL-6 como biomarcador en el lupus. Como objetivos específicos se plantea analizar la asociación de IL-6 con la actividad clínica mediante el SLEDAI, las manifestaciones clínicas, los parámetros inmunológicos e inflamatorios, y los biomarcadores serológicos de actividad conocidos, así como analizar si la IL-6 y los valores inflamatorios permiten determinar el tiempo medio en que se pueden iniciar las manifestaciones clínicas.Material y métodos: Se realizó un estudio observacional longitudinal prospectivo de una cohorte de 142 pacientes diagnosticados de LES según la clasificación EULAR/ACR, en el cual se incluían 587 determinaciones que presentaban la cuantificación de IL-6 entre 2017 y 2021.Se realizó un estudio descriptivo de todas las variables que forman parte del estudio, un análisis bivariante entre cada una de las 3 variables principales (IL-6, Autoacs anti-dsDNA y valores del complemento) y el resto de las variables, y un estudio de supervivencia para valorar si la IL-6 es un buen predictor de la enfermedad en el tiempo.Resultados: La IL-6 demostró tener una asociación significativa con la actividad del lupus, con los valores clínicos de artritis, miositis, neuritis, nefritis, serositis y afectación hematológica. Sin embargo, no se asoció con la afectación cutánea y las aftas. Respecto a los biomarcadores empleados en LES, tuvo asociación con los Autoanticuerpos anti-dsDNA, sin embargo no lo hubo para el complemento. Se demostró que una IL-6>5pg/ml tuvo una mayor actividad del LES en un tiempo medio inferior a 6 meses.Conclusiones: La IL-6 parece ser un buen marcador de actividad en diversas manifestaciones clínicas y analíticas del LES, y predictor en el tiempo.<br /

COVID GEAS: COVID-19 National Survey in Patients With Systemic Autoimmune Diseases

COVID-19 infection; Corticosteroids; Systemic autoimmune diseaseInfección por COVID-19; Corticosteroides; Enfermedad autoinmune sistémicaInfecció per COVID-19; Corticosteroides; Malaltia autoimmune sistèmicaObjectives: COVID-19 outcomes in population with systemic autoimmune diseases (SAD) remain poorly understood. The aim was to examine demographic and clinical factors associated with COVID-19 infection in people with rheumatic disease. Methods: Two phases cross-sectional survey of individuals with rheumatic disease in April 2020 and October 2020. COVID infection, severity of disease, age, sex, smoking status, underlying rheumatic disease diagnosis, comorbidities and rheumatic disease medications taken immediately prior to infection were analyzed. Results: A total of 1,529 individuals with autoimmunity disease diagnosis were included. Out of 50 positive patients, 21 required telephone medical assistance, 16 received assessment by primary care physician, 9 were evaluated in Emergency Department and 4 patient required hospitalization. Multivariate analysis was performed without obtaining differences in any of the systemic autoimmune diseases. Regarding the treatments, significant differences were found (p 0.011) in the treatment with anti-TNF-alpha agents with OR 3.422 (1.322–8.858) and a trend to significance (p 0.094) was observed in patients receiving mycophenolate treatment [OR 2.016 (0.996–4-081)]. Conclusions: Anti-TNF-alpha treatment was associated with more than 3-fold risk of suffering from SARS-CoV-2 infection, although in all cases infection was mild. Cumulative incidence in patients with SAD was up to 5 times higher than general population but with great differences between autoimmune diseases.This work was supported by SEMAIS

Intracellular delivery of biologically-active fungal metabolite gliotoxin using magnetic nanoparticles

Gliotoxin (GT), a secondary metabolite produced by Aspergillus molds, has been proposed as a potential anti-tumor agent. Here we have developed a nanoparticle approach to enhance delivery of GT in tumor cells and establish a basis for its potential use as therapeutical drug. GT bound to magnetic nanoparticles (MNPs) retained a high anti-tumor activity, correlating with efficient intracellular delivery, which was increased in the presence of glucose. Our results show that the attachment of GT to MNPs by covalent bonding enhances intracellular GT delivery without affecting its biological activity. This finding represents the first step to use this potent anti-tumor agent in the treatment of cancer

Real-time intracellular temperature imaging using lanthanide-bearing polymeric micelles

Measurement of thermogenesis in individual cells is a remarkable challenge due to the complexity of the biochemical environment (such as pH and ionic strength) and to the rapid and yet not well-understood heat transfer mechanisms throughout the cell. Here, we present a unique system for intracellular temperature mapping in a fluorescence microscope (uncertainty of 0.2 K) using rationally designed luminescent Ln3+-bearing polymeric micellar probes (Ln = Sm, Eu) incubated in breast cancer MDA-MB468 cells. Two-dimensional (2D) thermal images recorded increasing the temperature of the cells culture medium between 296 and 304 K shows inhomogeneous intracellular temperature progressions up to ∼20 degrees and subcellular gradients of ∼5 degrees between the nucleolus and the rest of the cell, illustrating the thermogenic activity of the different organelles and highlighting the potential of this tool to study intracellular processes.publishe

Importance of TRAIL Molecular Anatomy in Receptor Oligomerization and Signaling. Implications for Cancer Therapy

(TNF)-related apoptosis-inducing ligand (TRAIL) is able to activate the extrinsic apoptotic pathway upon binding to DR4/TRAIL-R1 and/or DR5/TRAIL-R2 receptors. Structural data indicate that TRAIL functions as a trimer that can engage three receptor molecules simultaneously, resulting in receptor trimerization and leading to conformational changes in TRAIL receptors. However, receptor conformational changes induced by the binding of TRAIL depend on the molecular form of this death ligand, and not always properly trigger the apoptotic cascade. In fact, TRAIL exhibits a much stronger pro-apoptotic activity when is found as a transmembrane protein than when it occurs as a soluble form and this enhanced biological activity is directly linked to its ability to cluster TRAIL receptors in supra-molecular structures. In this regard, cells involved in tumor immunosurveillance, such as activated human T cells, secrete endogenous TRAIL as a transmembrane protein associated with lipid microvesicles called exosomes upon T-cell reactivation. Consequently, it seems clear that a proper oligomerization of TRAIL receptors, which leads to a strong apoptotic signaling, is crucial for inducing apoptosis in cancer cells upon TRAIL treatment. In this review, the current knowledge of oligomerization status of TRAIL receptors is discussed as well as the implications for cancer treatment when using TRAIL-based therapies