TRAIL receptors promote constitutive and inducible IL-8 secretion in non-small cell lung carcinoma

Interleukin-8 (IL-8/CXCL8) is a pro-angiogenic and pro-inflammatory chemokine that plays a role in cancer development. Non-small cell lung carcinoma (NSCLC) produces high amounts of IL-8, which is associated with poor prognosis and resistance to chemo-radio and immunotherapy. However, the signaling pathways that lead to IL-8 production in NSCLC are unresolved. Here, we show that expression and release of IL-8 are regulated autonomously by TRAIL death receptors in several squamous and adenocarcinoma NSCLC cell lines. NSCLC constitutively secrete IL-8, which could be further enhanced by glucose withdrawal or by treatment with TRAIL or TNF alpha. In A549 cells, constitutive and inducible IL-8 production was dependent on NF-kappa B and MEK/ERK MAP Kinases. DR4 and DR5, known regulators of these signaling pathways, participated in constitutive and glucose deprivation-induced IL-8 secretion. These receptors were mainly located intracellularly. While DR4 signaled through the NF-kappa B pathway, DR4 and DR5 both regulated the ERK-MAPK and Akt pathways. FADD, caspase-8, RIPK1, and TRADD also regulated IL-8. Analysis of mRNA expression data from patients indicated that IL-8 transcripts correlated with TRAIL, DR4, and DR5 expression levels. Furthermore, TRAIL receptor expression levels also correlated with markers of angiogenesis and neutrophil infiltration in lung squamous carcinoma and adenocarcinoma. Collectively, these data suggest that TRAIL receptor signaling contributes to a pro-tumorigenic inflammatory signature associated with NSCLC

Chemokine C-C motif ligand 2 overexpression drives tissue-specific metabolic responses in the liver and muscle of mice

Chemokine (C-C motif) ligand 2 (CCL2) has been associated with chronic metabolic diseases. We aimed to investigate whether Ccl2 gene overexpression is involved in the regulation of signaling pathways in metabolic organs. Biochemical and histological analyses were used to explore tissue damage in cisgenic mice that overexpressed the Ccl2 gene. Metabolites from energy and one-carbon metabolism in liver and muscle extracts were measured by targeted metabolomics. Western blot analysis was used to explore the AMP-activated protein kinase (AMPK) and mammalian target of rapamycin pathways. Ccl2 overexpression resulted in steatosis, decreased AMPK activity and altered mitochondrial dynamics in the liver. These changes were associated with decreased oxidative phosphorylation and alterations in the citric acid cycle and transmethylation. In contrast, AMPK activity and its downstream mediators were increased in muscle, where we observed an increase in oxidative phosphorylation and increased concentrations of different metabolites associated with ATP synthesis. In conclusion, Ccl2 overexpression induces distinct metabolic alterations in the liver and muscle that affect mitochondrial dynamics and the regulation of energy sensors involved in cell homeostasis. These data suggest that CCL2 may be a therapeutic target in metabolic diseases

The role of chemokine (c-c motif) ligand 2 in inflammation, oxidative stress, aging and metabolism

El metabolisme i el sistema immunològic estan estretament relacionats i les seves interaccions juguen un paper important en l'homeòstasi sistèmica. L'activació, proliferació, diferenciació i polarització de les cèl·lules immunitàries representen un important estrès metabòlic, que pot comprometre el metabolisme cel·lular, l'homeòstasi i les necessitats energètiques. Aquestes condicions poden promoure l'estrès metabòlic crònic i les anomalies metabòliques en patologies no immunes. No obstant això, encara no queda clar com aquests canvis en el perfil immunològic afecten el metabolisme sistèmic. Cada vegada hi ha més proves de que les quimiocines exerceixen un paper crucial en tots aquests processos. Especialment, l'augment de la quimiocina C-C motif ligand 2 (CCL2) en malalties metabòliques suggereix la possibilitat que aquesta jugui un rol en la regulació del metabolisme. La pregunta clau és si l'augment de CCL2 és la causa o conseqüència del problema. Per aquesta raó, explorem l'efecte de la deficiència de CCL2 en el metabolisme de ratolins amb hiperlipèmia, esteatosis hepàtica i síndrome metabòlica. A més, es va investigar si aquest efecte podria estar condicionat per la dieta. Els resultats obtinguts es presenten en el primer estudi. Aquests resultats obren la porta a altres preguntes. Per exemple, la funció CCL2 va més enllà de la seva capacitat d'atraure cèl·lules immunitàries? Pot aquesta quimiocina afectar el metabolisme energètic? Per a respondre a aquestes preguntes, generem ratolins CCL2 cisgenic. Els resultats es presenten en l'estudi 2. En l'estudi 3 es va avaluar l'efecte de la sobre-expressió de CCL2 en un model de ratolí de la síndrome de progèria de Hutchinson-Gilford, un model d'envelliment accelerat. La cerca de dianes terapèutiques per al tractament de malalties metabòliques és un punt important en les recerques actuals. Els resultats d'aquesta tesi suggereixen que la CCL2 podria ser una diana terapèutica important en diferents malalties metabòliques.El metabolismo y el sistema inmunológico están estrechamente relacionados y sus interacciones juegan un papel importante en la homeostasis sistémica. La activación, proliferación, diferenciación y polarización de las células inmunitarias representan un importante estrés metabólico, que puede comprometer el metabolismo celular, la homeostasis y las necesidades energéticas. Estas condiciones pueden promover el estrés metabólico crónico y las anomalías metabólicas en patologías no inmunes. Sin embargo, aún no está claro cómo estos cambios en el perfil inmunológico afectan al metabolismo sistémico. Cada vez hay más pruebas de que las quimiocinas desempeñan un papel crucial en todos estos procesos. Especialmente, el aumento de la quimiocina C-C motif ligand 2 (CCL2) en enfermedades metabólicas sugiere la posibilidad de que esta juegue un papel en la regulación del metabolismo. La pregunta clave es si el aumento de CCL2 es la causa o consecuencia del problema. Por esta razón, exploramos el efecto de la deficiencia de CCL2 en el metabolismo de ratones con hiperlipidemia, esteatosis hepática y síndrome metabólico. Además, se investigó si ese efecto podría estar condicionado por la dieta. Los resultados obtenidos se presentan en el primer estudio. Estos resultados abren la puerta a otras preguntas. Por ejemplo, ¿la función de CCL2 va más allá de su capacidad de atraer células inmunitarias? ¿Puede esta quimiocina afectar el metabolismo energético? Para responder a estas preguntas, generamos ratones CCL2 cisgénicos. Los resultados se presentan en el estudio 2. En el estudio 3 se evaluó el efecto de la sobreexpresión de CCL2 en un modelo de ratón del síndrome de progeria de Hutchinson-Gilford, un modelo de envejecimiento acelerado. La búsqueda de dianas terapéuticas para el tratamiento de enfermedades metabólicas es un punto importante en las investigaciones actuales. Los resultados de esta tesis sugieren que CCL2 podría ser una diana terapéutica importante en diferentes enfermedades metabólicas.Metabolism and immune system are closely interconnected and their interactions play an important role in whole-body homeostasis. The activation, proliferation, differentiation and polarization of the immune cells represent significant metabolic stress, which can compromise the cellular metabolism, homeostasis and energetics requirements. These conditions can promote the chronic metabolic stress and metabolic abnormalities in non-immune pathologies. Nevertheless, how these changes in the immunological profile affect systemic metabolism are still not clear. Growing evidences support that chemokines play a crucial role in all these processes. Specially, the increase of chemokine C-C motif ligand 2 (CCL2) in metabolic diseases suggests the possibility of this chemokine to play a systemic role in the regulation of metabolism. The key question is if increase of CCL2 is the cause or consequence of the problem. For this reason, we explored the effect of CCL2 ablation in the metabolism of mice with a background of hyperlipidemia, hepatic steatosis and metabolic syndrome. In addition, we investigated whether that effect might be conditioned by diet. Obtained results are presented in the first study. These results open a brief to other questions. For example, does CCL2 function go further to its chemoattracting capacity? Can this chemokine affect the systemic energy metabolism? To answer these questions, we generated targeted CCL2 cisgenic mice, which overexpressed CCL2 in all tissues and results are presented in study 2. In study 3 we evaluated the effect of CCL2 overexpression in a mice model of Hutchinson-Gilford progeria syndrome, a model of accelerated aging. The finding of therapeutic targets for the treatment of metabolic diseases is an important point in current investigations in our research group. The conclusions of this thesis suggest that CCL2 could be an important therapeutic target in different metabolic diseases

The role of chemokine (c-c motif) ligand 2 in inflammation, oxidative stress, aging and metabolism

El metabolisme i el sistema immunològic estan estretament relacionats i les seves interaccions juguen un paper important en l'homeòstasi sistèmica. L'activació, proliferació, diferenciació i polarització de les cèl·lules immunitàries representen un important estrès metabòlic, que pot comprometre el metabolisme cel·lular, l'homeòstasi i les necessitats energètiques. Aquestes condicions poden promoure l'estrès metabòlic crònic i les anomalies metabòliques en patologies no immunes. No obstant això, encara no queda clar com aquests canvis en el perfil immunològic afecten el metabolisme sistèmic. Cada vegada hi ha més proves de que les quimiocines exerceixen un paper crucial en tots aquests processos. Especialment, l'augment de la quimiocina C-C motif ligand 2 (CCL2) en malalties metabòliques suggereix la possibilitat que aquesta jugui un rol en la regulació del metabolisme. La pregunta clau és si l'augment de CCL2 és la causa o conseqüència del problema. Per aquesta raó, explorem l'efecte de la deficiència de CCL2 en el metabolisme de ratolins amb hiperlipèmia, esteatosis hepàtica i síndrome metabòlica. A més, es va investigar si aquest efecte podria estar condicionat per la dieta. Els resultats obtinguts es presenten en el primer estudi. Aquests resultats obren la porta a altres preguntes. Per exemple, la funció CCL2 va més enllà de la seva capacitat d'atraure cèl·lules immunitàries? Pot aquesta quimiocina afectar el metabolisme energètic? Per a respondre a aquestes preguntes, generem ratolins CCL2 cisgenic. Els resultats es presenten en l'estudi 2. En l'estudi 3 es va avaluar l'efecte de la sobre-expressió de CCL2 en un model de ratolí de la síndrome de progèria de Hutchinson-Gilford, un model d'envelliment accelerat. La cerca de dianes terapèutiques per al tractament de malalties metabòliques és un punt important en les recerques actuals. Els resultats d'aquesta tesi suggereixen que la CCL2 podria ser una diana terapèutica important en diferents malalties metabòliques.El metabolismo y el sistema inmunológico están estrechamente relacionados y sus interacciones juegan un papel importante en la homeostasis sistémica. La activación, proliferación, diferenciación y polarización de las células inmunitarias representan un importante estrés metabólico, que puede comprometer el metabolismo celular, la homeostasis y las necesidades energéticas. Estas condiciones pueden promover el estrés metabólico crónico y las anomalías metabólicas en patologías no inmunes. Sin embargo, aún no está claro cómo estos cambios en el perfil inmunológico afectan al metabolismo sistémico. Cada vez hay más pruebas de que las quimiocinas desempeñan un papel crucial en todos estos procesos. Especialmente, el aumento de la quimiocina C-C motif ligand 2 (CCL2) en enfermedades metabólicas sugiere la posibilidad de que esta juegue un papel en la regulación del metabolismo. La pregunta clave es si el aumento de CCL2 es la causa o consecuencia del problema. Por esta razón, exploramos el efecto de la deficiencia de CCL2 en el metabolismo de ratones con hiperlipidemia, esteatosis hepática y síndrome metabólico. Además, se investigó si ese efecto podría estar condicionado por la dieta. Los resultados obtenidos se presentan en el primer estudio. Estos resultados abren la puerta a otras preguntas. Por ejemplo, ¿la función de CCL2 va más allá de su capacidad de atraer células inmunitarias? ¿Puede esta quimiocina afectar el metabolismo energético? Para responder a estas preguntas, generamos ratones CCL2 cisgénicos. Los resultados se presentan en el estudio 2. En el estudio 3 se evaluó el efecto de la sobreexpresión de CCL2 en un modelo de ratón del síndrome de progeria de Hutchinson-Gilford, un modelo de envejecimiento acelerado. La búsqueda de dianas terapéuticas para el tratamiento de enfermedades metabólicas es un punto importante en las investigaciones actuales. Los resultados de esta tesis sugieren que CCL2 podría ser una diana terapéutica importante en diferentes enfermedades metabólicas.Metabolism and immune system are closely interconnected and their interactions play an important role in whole-body homeostasis. The activation, proliferation, differentiation and polarization of the immune cells represent significant metabolic stress, which can compromise the cellular metabolism, homeostasis and energetics requirements. These conditions can promote the chronic metabolic stress and metabolic abnormalities in non-immune pathologies. Nevertheless, how these changes in the immunological profile affect systemic metabolism are still not clear. Growing evidences support that chemokines play a crucial role in all these processes. Specially, the increase of chemokine C-C motif ligand 2 (CCL2) in metabolic diseases suggests the possibility of this chemokine to play a systemic role in the regulation of metabolism. The key question is if increase of CCL2 is the cause or consequence of the problem. For this reason, we explored the effect of CCL2 ablation in the metabolism of mice with a background of hyperlipidemia, hepatic steatosis and metabolic syndrome. In addition, we investigated whether that effect might be conditioned by diet. Obtained results are presented in the first study. These results open a brief to other questions. For example, does CCL2 function go further to its chemoattracting capacity? Can this chemokine affect the systemic energy metabolism? To answer these questions, we generated targeted CCL2 cisgenic mice, which overexpressed CCL2 in all tissues and results are presented in study 2. In study 3 we evaluated the effect of CCL2 overexpression in a mice model of Hutchinson-Gilford progeria syndrome, a model of accelerated aging. The finding of therapeutic targets for the treatment of metabolic diseases is an important point in current investigations in our research group. The conclusions of this thesis suggest that CCL2 could be an important therapeutic target in different metabolic diseases

Nutrients in Energy and One-Carbon Metabolism: Learning from Metformin Users

Metabolic vulnerability is associated with age-related diseases and concomitant co-morbidities, which include obesity, diabetes, atherosclerosis and cancer. Most of the health problems we face today come from excessive intake of nutrients and drugs mimicking dietary effects and dietary restriction are the most successful manipulations targeting age-related pathways. Phenotypic heterogeneity and individual response to metabolic stressors are closely related food intake. Understanding the complexity of the relationship between dietary provision and metabolic consequences in the long term might provide clinical strategies to improve healthspan. New aspects of metformin activity provide a link to many of the overlapping factors, especially the way in which organismal bioenergetics remodel one-carbon metabolism. Metformin not only inhibits mitochondrial complex 1, modulating the metabolic response to nutrient intake, but also alters one-carbon metabolic pathways. Here, we discuss findings on the mechanism(s) of action of metformin with the potential for therapeutic interpretations

Galectin-3 in Peripheral Artery Disease. Relationships with Markers of Oxidative Stress and Inflammation

Galectin-3 is a modulator of oxidative stress, inflammation, and fibrogenesis involved in the pathogenesis of vascular diseases. The present study sought to characterize, in patients with peripheral artery disease (PAD), the localization of galectin-3 in arterial tissue, and to analyze the relationships between the circulating levels of galectin-3 and oxidative stress and inflammation. It also sought to compare the diagnostic accuracy of galectin-3 with that of other biochemical markers of this disease. We analyzed femoral or popliteal arteries from 50 PAD patients, and four control arteries. Plasma from 86 patients was compared with that from 72 control subjects. We observed differences in the expression of galectin-3 in normal arteries, and arteries from patients with PAD, with a displacement of the expression from the adventitia to the media, and the intima. In addition, plasma galectin-3 concentration was increased in PAD patients, and correlated with serologic markers of oxidative stress (F2-isoprostanes), and inflammation [chemokine (C−C motif) ligand 2, C-reactive protein, β-2-microglobulin]. We conclude that the determination of galectin-3 has good diagnostic accuracy in the assessment of PAD and compares well with other analytical parameters currently in use

Plasma Energy-Balance Metabolites Discriminate Asymptomatic Patients with Peripheral Artery Disease

Peripheral artery disease (PAD) is a common disease affecting 20–25% of population over 60 years old. Early diagnosis is difficult because symptoms only become evident in advanced stages of the disease. Inflammation, impaired metabolism, and mitochondrial dysfunction predispose to PAD, which is normally associated with other highly prevalent and related conditions, such as diabetes, dyslipidemia, and hypertension. We have measured energy-balance-associated metabolite concentrations in the plasma of PAD patients segregated by the severity of the disease and in plasma of healthy volunteers using a quantitative and targeted metabolomic approach. We found relevant associations between several metabolites (3-hydroxybutirate, aconitate, (iso)citrate, glutamate, and serine) with markers of oxidative stress and inflammation. Metabolomic profiling also revealed that (iso)citrate and glutamate are metabolites with high ability to discriminate between healthy participants and PAD patients without symptoms. Collectively, our data suggest that metabolomics provide significant information on the pathogenesis of PAD and useful biomarkers for the diagnosis and assessment of progression

Metformin Potentiates the Benefits of Dietary Restraint: A Metabolomic Study

Prevention of the metabolic consequences of a chronic energy-dense/high-fat diet (HFD) represents a public health priority. Metformin is a strong candidate to be incorporated in alternative therapeutic approaches. We used a targeted metabolomic approach to assess changes related to the multi-faceted metabolic disturbances provoked by HFD. We evaluated the protective effects of metformin and explored how pro-inflammatory and metabolic changes respond when mice rendered obese, glucose-intolerant and hyperlipidemic were switched to diet reversal with or without metformin. Mice treated with metformin and diet-reversal showed a dramatically improved protection against HFD-induced hepatic steatosis, a beneficial effect that was accompanied by a lowering of liver-infiltrating pro-inflammatory macrophages and lower release of pro-inflammatory cytokines. Metformin combined with diet reversal promoted effective weight loss along with better glucose control, lowered levels of circulating cholesterol and triglycerides, and reduced adipose tissue content. Our findings underscored the ability of metformin to target the contribution of branched chain amino acids to adipose tissue metabolism while suppressing mitochondrial-dependent biosynthesis in hepatic tissue. The relationship between adipose tissue and liver might provide clinical potential for combining metformin and dietary modifications to protect against the metabolic damage occurring upon excessive dietary fat intake

Immunohistochemical Analysis of Paraoxonases and Chemokines in Arteries of Patients with Peripheral Artery Disease

Oxidative damage to lipids and lipoproteins is implicated in the development of atherosclerotic vascular diseases, including peripheral artery disease (PAD). The paraoxonases (PON) are a group of antioxidant enzymes, termed PON1, PON2, and PON3 that protect lipoproteins and cells from peroxidation and, as such, may be involved in protection against the atherosclerosis process. PON1 inhibits the production of chemokine (C–C motif) ligand 2 (CCL2) in endothelial cells incubated with oxidized lipoproteins. PON1 and CCL2 are ubiquitously distributed in tissues, and this suggests a joint localization and combined systemic effect. The aim of the present study has been to analyze the quantitative immunohistochemical localization of PON1, PON3, CCL2 and CCL2 receptors in a series of patients with severe PAD. Portions of femoral and/or popliteal arteries from 66 patients with PAD were obtained during surgical procedures for infra-inguinal limb revascularization. We used eight normal arteries from donors as controls. PON1 and PON3, CCL2 and the chemokine-binding protein 2, and Duffy antigen/chemokine receptor, were increased in PAD patients. There were no significant changes in C–C chemokine receptor type 2. Our findings suggest that paraoxonases and chemokines play an important role in the development and progression of atherosclerosis in peripheral artery disease

Effect of radiotherapy on activity and concentration of serum paraoxonase-1 in breast cancer patients.

Paraoxonase-1 (PON1) is an intra-cellular antioxidant enzyme found also in the circulation associated with high-density lipoproteins. The activity of this enzyme has been shown to be decreased in breast cancer (BC) patients. The aims of our study were to investigate the changes produced by radiotherapy (RT) on activity and concentration of serum PON1 in BC patients, and to evaluate the observed variations in relation to clinical and pathological characteristics of patients and tumors, and the response to treatment. We studied 200 women with BC who were scheduled to receive RT following excision of the tumor. Blood for analyses was obtained before and after the irradiation procedure. The control group was composed of 200 healthy women. Relative to control, BC patients had significantly lower serum PON1 activities pre-RT, while PON1 concentrations were at similar levels. RT was associated with a significant increase in serum PON1 activities and concentrations. We observed significant differences in serum PON1 concentrations post-RT between patients with luminal A or luminal B tumors. Serum PON1 concentration post-RT was markedly lower in BC patients with metastases. We conclude that benefit from RT accrues to the BC patients not only through its direct effect on cancer cells but also indirectly by improving the organism's anti-oxidant defense mechanisms. In addition, our preliminary evidence suggests that the measurement of serum PON1 concentration post-RT could be an efficient prognostic biomarker, and may be used as an index of the efficacy of the RT