Search

A combination of molecular modeling and structure−activity relationship studies has been used to fine-tune CB2 selectivity in the chromenopyrazole ring, a versatile CB1/CB2 cannabinoid scaffold. Thus, a series of 36 new derivatives covering a wide range of structural diversity has been synthesized, and docking studies have been performed for some of them. Biological evaluation of the new compounds includes, among others, cannabinoid binding assays, functional studies, and surface plasmon resonance measurements. The most promising compound [43 (PM226)], a selective and potent CB2 agonist isoxazole derivative, was tested in the acute phase of Theiler’s murine encephalomyelitis virus-induced demyelinating disease (TMEV-IDD), a well established animal model of primary progressive multiple sclerosis. Compound 43 dampened neuroinflammation by reducing microglial activation in the TMEV, Ministerio de Economía, Comercio y Empresa, Comunidad de Madrid, Depto. de Bioquímica y Biología Molecular, Fac. de Medicina, TRUE, pub

Chromenopyrazolediones have been designed and synthesized as anticancer agents using the multi biological target concept that involves quinone cytotoxicity and cannabinoid antitumor properties. In cell cytotoxicity assays, these chromenopyrazolediones have antiproliferative activity against human prostate cancer and hepatocellular carcinoma. It has been shown that the most potent, derivative 4 (PM49), inhibits prostate LNCaP cell viability (IC50 ¼ 15 mM) through a mechanism involving oxidative stress, PPARg receptor and partially CB1 receptor. It acts on prostate cell growth by causing G0/G1 phase arrest and triggering apoptosis as assessed by flow cytometry measurements. In the in vivo treatment, compound 4 at 2 mg/kg, blocks the growth of LNCaP tumors and reduces the growth of PC-3 tumors generated in mice. These studies suggest that 4 is a good potential anticancer agent against hormone sensitive prostate cancer, Depto. de Bioquímica y Biología Molecular, Fac. de Medicina, TRUE, pub

The orphan G protein-coupled receptor GPR55 has been proposed as a novel receptor of the endocannabinoid system. However, the validity of this categorization is still under debate mainly because of the lack of potent and selective agonists and antagonists of GPR55. Binding assays are not yet available for GPR55 screening, and discrepancies in GPR55 mediated signaling pathways have been reported. In this context, we have designed and synthesized novel GPR55 ligands based on a chromenopyrazole scaffold. Appraisal of GPR55 activity was accomplished using a label-free cell-impedance-based assay in hGPR55-HEK293 cells. The real-time impedance responses provided an integrative assessment of the cellular consequence to GPR55 stimulation taking into account the different possible signaling pathways. Potent GPR55 partial agonists (14b, 18b, 19b, 20b, and 21−24) have been identified; one of them (14b) being selective versus classical cannabinoid receptors. Upon antagonist treatment, chromenopyrazoles 21−24 inhibited lysophosphatidylinositol (LPI) effect. One of these GPR55 antagonists (21) is fully selective versus classic cannabinoid receptors. Compared to LPI, the predicted physicochemical parameters of the new compounds suggest a clear pharmacokinetic improvement, Ministerio de Economía, Comercio y Empresa, Comunidad de Madrid, Depto. de Bioquímica y Biología Molecular, Fac. de Medicina, TRUE, pub

Cannabinoids have emerged as promising neuroprotective agents due to their capability to activate specific targets, which are involved in the control of neuronal homeostasis and survival. Specifically, those ligands that selectively target and activate the CB2 receptor may be useful for their anti inflammatory and neuroprotective properties in various neurological disorders, with the advantage of being devoid of psychotropic effects associated with the activation of CB1 receptors. The aim of this work has been to investigate the neuroprotective properties of 7-(1,1-dimethylheptyl)-4,4-dimethyl-9- methoxychromeno[3,4-d]isoxazole (PM226), a compound derived from a series of chromeno-isoxazoles and −pyrazoles, which seems to have a promising profile related to the CB2 receptor. The compound binds selectively to this receptor with an affinity in the nanomolar range (Ki = 12.8 ± 2.4 nM). It has negli gible affinity for the CB1 receptor (Ki > 40000 nM) and no activity at the GPR55. PM226 was also evaluated in GTP S binding assays specific to the CB2 receptor showing agonist activity (EC50 = 38.67 ± 6.70 nM). In silico analysis of PM226 indicated that it has a good pharmacokinetic profile and a predicted abil ity to cross the blood-brain barrier. Next, PM226 was investigated in an in vitro model to explore its anti-inflammatory/neuroprotective properties. Conditioned media were collected from LPS-stimulated cultures of BV2 microglial cell line in the absence or presence of different doses of PM226, and then media were added to cultured M213-2O neuronal cells to record their influence on cell viability evaluated using MTT assays. As expected, cell viability was significantly reduced by the exposure to these conditioned media, while the addition of PM226 attenuated this reduction in a dose-dependent manner. This effect was reversed by co-incubating with the CB2 antagonist SR144528, thus confirming the involvement of CB2 receptors, whereas the addition of PM226 to neuronal cu, Ministerio de Economía, Comercio y Empresa, Centro de Investigación Biomédica en Red (CIBERNED), Comunidad de Madrid, Depto. de Bioquímica y Biología Molecular, Fac. de Medicina, TRUE, pub

[ES] Los nódulos tiroideos son la patología endocrina más frecuente, diagnosticándose malignidad en una pequeña proporción (∼15 %). La incidencia del cáncer de tiroides se ha triplicado en España en los últimos 25 años. Así mismo, afecta tres veces más a mujeres que a hombres y se incrementa su frecuencia con la edad, sobre todo a partir de los 40 años. En general, son tumores de gran pronóstico, con tasas de supervivencia cercanas al 100 %. Sin embargo, debido a la alta prevalencia de los nódulos en la población es necesario un triaje exhaustivo. La punción por aspiración con aguja fina es la prueba no quirúrgica de referencia para el diagnóstico inicial de los nódulos tiroideos. Sin embargo, el 25-30 % de todas las punciones son designadas como indeterminadas y dada que la posibilidad de malignidad, muchos pacientes suelen ser derivados a cirugía. Tras la operación, solo el 25 % de las lesiones resultan malignas, por lo que el riesgo de cáncer no es lo suficientemente alto como para respaldar definitivamente la cirugía. En retrospectiva, estas tiroidectomías son innecesarias y generan complicaciones que los pacientes padecerán durante toda su vida. Por lo tanto, existe una necesidad urgente de desarrollar métodos efectivos para discernir benignidad de malignidad en las lesiones foliculares de tiroides. El desarrollo de biomarcadores seguros y rentables podría ayudar en la toma de decisiones del médico tratante para manejar con mayor certeza los nódulos tiroideos. Si bien durante mucho tiempo se ha tratado de establecer el valor diagnóstico de las diferentes alteraciones genéticas ninguna parece ser totalmente resolutiva. La metilación del ADN es una de las principales modificaciones epigenéticas y su papel en el inicio, desarrollo y metástasis de cánceres humanos ha sido ampliamente descrito, incluido en cáncer de tiroides. Dado que las firmas de metilación del ADN pueden ser indicativas de la plasticidad molecular que surge en la tumorigénesis temprana, en este e, [EN] Thyroid nodules are the most common encountered thyroid endocrine pathology, being unfrequently diagnosed as malignant (∼15%). The incidence of thyroid cancer has tripled in Spain in the last 25 years. Likewise, it affects three times more women than men and its frequency increases with age, especially after the age of 40. In general, they are tumors with a great prognosis, with survival rates close to 100%. However, due to the high prevalence of nodules in the population, an exhaustive triage is necessary. Fine needle aspiration puncture is the gold standard non-surgical test for the initial diagnosis of thyroid nodules. However, 25-30% of all cytologies are designated as indeterminate and given the possibility of malignancy, many patients are often referred for surgery. After the operation, only 25% of lesions result as cancer, so the risk of malignancy is not high enough to definitively support thyroidectomy. In hindsight, these thyroidectomies are necessary and lead to complications that patients will endure for their entire lives. Therefore, there is an urgent need to develop effective methods to discern benign from malignant follicular-patterned thyroid lesions. Safe and cost-effective biomarkers could help in the decision-making of the treating physician to manage thyroid nodules with greater certainty. Although many attempts have been made to establish the diagnostic value of the different genetic alterations, none of them seems to be fully resolvable. DNA methylation is one of the main epigenetic modifications and its role in the initiation, development and metastasis of human cancers has been widely described, including thyroid cancer. Since DNA methylation signatures may be indicative of molecular plasticity that arises in early tumorigenesis, this study postulated that this epigenetic mark could be used as a marker for discrimination in difficult-to-diagnose cases. Initially, mutation analysis was performed by whole-exome sequencing in a retrospecti

Type 1 diabetes is a chronic autoimmune disease which results in inefficient regulation of glucose homeostasis and can lead to different vascular comorbidities through life. In this study we aimed to analyse the circulating miRNA expression profile of patients with type 1 diabetes, and with no other associated pathology. For this, fasting plasma was obtained from 85 subjects. Next generation sequencing analysis was firstly performed to identify miRNAs that were differentially expressed between groups (20 patients vs. 10 controls). hsa-miR-1-3p, hsa-miR-200b-3p, hsa-miR-9-5p, and hsa-miR-1200 expression was also measured by Taqman RT-PCR to validate the observed changes (34 patients vs. 21 controls). Finally, through a bioinformatic approach, the main pathways affected by the target genes of these miRNAs were studied. Among the studied miRNAs, hsa-miR-1-3p expression was found significantly increased in patients with type 1 diabetes compared to controls, and positively correlated with glycated haemoglobin levels. Additionally, by using a bioinformatic approach, we could observe that changes in hsa-miR-1-3p directly affect genes involved in vascular development and cardiovascular pathologies. Our results suggest that, circulating hsa-miR-1-3p in plasma, together with glycaemic control, could be used as prognostic biomarkers in type 1 diabetes, helping to prevent the development of vascular complications in these patients.

The second Transatlantic Early Career Investigator (ECI) G Protein-Coupled Receptor (GPCR) Symposium was an online scientific meeting geared at young GPCR investigators, with the primary goal of expanding opportunities for sharing research and networking among trainees in North America and Europe. Here, we discuss the format of our meeting, its impact, and the challenges and opportunities facing meetings like it in the future.

descripción no proporcionada por scopus