Nutritional Immunotherapy: targeting immunometabolism with bioactive food compounds

La immunoteràpia nutricional es basa en fomentar la salut mitjançant el consum de compostos bioactius presents de manera natural en els aliments, com les procianidines, un tipus de flavonoid, i/o l’àcid docosahexaenoic (DHA), un àcid gras omega-3, optimitzant la funcionalitat del propi sistema immune i millorant així el seu paper com a responsable de la preservació de l’organisme enfront desestabilitzadors de la homeòstasis. La recerca que aquesta tesi abasta es centra en el paper del perfil nutricional en la regulació de la interacció immunitat-metabolisme (immunometabolisme). Amb aquesta finalitat, en la primera part del treball presentat en aquesta tesi, es va determinar si els macròfags, les cèl•lules efectores de la immunitat innata, poden percebre de manera diferencial la composició de la dieta. Hem demostrat que els compostos bioactius dels aliments modulen, a nivell molecular, l’activació dels macròfags. A més, aquest efecte immunomodulador és dependent del compost, on factors intrínsecs com les propietats químiques o nutritives són determinants per la seva bioactivitat. La segona part va tenir com a objectiu determinar el paper primordial del patró nutricional en la regulació de la interacció entre el sistema immune i el metabolisme. Utilitzant un model d’obesitat induïda per la dieta es va inferir que, mentre una dieta hipercalòrica provoca un deteriorament de l’immunometabolisme, el consum de bioactius pot enfortir-ne la relació. A la tercera part, es va analitzar el paper de les interaccions gen-dieta en l’expressió fenotípica dels trets associats amb la obesitat. Per aquest motiu, dos races de rates congènites, fenotípicament diferents, van ser sotmeses a un desequilibri immunometabòlic com a conseqüència de la ingesta d’una dieta hipercalòrica. Es va deduir que les interaccions entre els factors genètics i nutricionals són fonamentals per a la susceptibilitat d’un genotip a l’obesitat induïda per la dieta. Hem establert que el perfil nutricional és una eina poderosa per orientar la funcionalitat de l’eix immunometabòlic. A més, arribem a la conclusió que compostos bioactius presents en els aliments poden millorar-ne l’eficiència d’aquest eix, promovent així un estat saludable.La immunoterapia nutricional se basa en fomentar la salud mediante el consumo de componentes bioactivos presentes de forma natural en los alimentos, como las procianidinas, un tipo de flavonoides, o el ácido docosahexaenoico (DHA), un ácido graso omega-3, optimizando la funcionalidad del propio sistema inmune y mejorando así su papel como responsable de la preservación del organismo frente a desestabilizadores de la homeostasis. La investigación que engloba esta tesis se centra en el papel del perfil nutricional en la regulación de la interacción inmunidad-metabolismo (immunometabolismo). Con esta finalidad, en la primera parte del trabajo presentado es esta tesis, se determinó si los macrófagos, las células efectoras de la inmunidad innata, pueden percibir de manera diferencial la composición de la dieta. Hemos demostrado que los compuestos bioactivos de los alimentos modulan, a nivel molecular, la activación de los macrófagos. Además, este efecto immunomodulador es dependiente del compuesto, donde factores intrínsecos tales como sus propiedades químicas o nutritivas son determinantes para su bioactividad. La segunda parte tuvo como objetivo determinar el papel primordial del patrón nutricional en la regulación de la interacción entre el sistema inmune y el metabolismo. Utilizando un modelo de obesidad inducida por la dieta se infirió que, mientras una dieta hipercalórica provoca un deterioro del immunometabolismo, el consumo de bioactivos pueden fortalecer su estrecha relación. En la tercera parte, se analizó el papel de las interacciones gen-dieta en la expresión fenotípica de los rasgos asociados con la obesidad. Para ello, dos razas de ratas congénitas, fenotípicamente diferentes, fueron sometidas a un desequilibrio immunometabólico como consecuencia de la ingesta de una dieta hipercalórica. Se dedujo que las interacciones entre los factores genéticos y nutricionales son fundamentales para la susceptibilidad de un genotipo a la obesidad inducida por la dieta. Hemos establecido que el perfil nutricional es una herramienta poderosa para orientar la funcionalidad del eje immunometabólico. Además, llegamos a la conclusión que compuestos bioactivos presentes en los alimentos pueden mejorar la eficiencia de este eje, promoviendo así un estado saludable.Nutritional immunotherapy is based on promoting health through the dietary intake of natural bioactive compounds found in food, such as the procyanidins and docosahexaenoic n-3 polyunsaturated fatty acid (DHA), optimising the functionality of the host’s own immune system and improving its role in the preservation of the body against destabilisers of homeostasis. The research that this thesis encompasses is focused on the role of the nutritional profile in the regulation of immunometabolism. To accomplish this purpose, in the first part of the work presented in this thesis, we determined whether macrophages, the effector cells of innate immunity, could differentially sense dietary composition. We demonstrated that bioactive food compounds modulate, at the molecular level, the functionality of macrophages by hindering macrophage activation. Furthermore, this immunomodulatory effect is compound-dependent, relying on the intrinsic factors of each compound, such as chemical and nutritional properties, to determine its bioactivity. The second part was aimed at determining the role of the dietary pattern within the complex crosstalk of immunity and metabolism using an in vivo model of diet-induced obesity. We determined that immunometabolic regulation depends on the nutritional profile. While a diet based on foods with a high energy content can weaken immunometabolism, the presence of bioactive foods can strengthen the relationship. Within the third part, we evaluated the role of gene-diet interactions in the phenotypic expression of obesity-related complex traits. Using a diet-induced obesity model, two distinct genetic backgrounds (phenotypically different inbred rat strains) were immunometabolically challenged. Our results revealed that interactions between genetic and dietary factors are fundamental for the susceptibility of a genotype to diet-induced obesity. We established that the nutritional profile is a powerful tool to target the functionality of the immunometabolic axis. We further concluded that bioactive compounds present in food improve the efficiency of this axis, thereby promoting a healthy state

Glycoprotein and Lipoprotein Profiles Assessed by 1H-NMR and Its Relation to Ascending Aortic Dilatation in Bicuspid Aortic Valve Disease

Bicuspid aortic valve; Glycoprotein; Lipoprotein metabolismVàlvula aòrtica bicúspide; Glicoproteïna; Metabolisme de les lipoproteïnesVálvula aórtica bicúspide; Glicoproteína; Metabolismo de las lipoproteínasIntroduction: The bicuspid aortic valve (BAV) confers a high risk of ascending aorta dilatation (AAoD), although its progression seems highly variable. Furthermore, the implication of lipoprotein metabolism and inflammation in the mechanisms that underlie AAoD is not fully established. The aim of this study consisted of evaluating the impact of the lipoprotein and glycoprotein profiles in AAOD as well as its progression in BAV aortopathy. Methods: Using 1H-nuclear magnetic resonance (1H-NMR), we analyzed and compared the lipoprotein and glycoprotein profiles of plasma samples from 152 BAV patients with dilated and nondilated ascending aorta. Additionally, these profiles were also compared for 119 of these patients who were prospectively followed-up clinically and by echocardiography in the long-term (5 years). Ascending aorta dilation velocity (mm/year) was calculated for this analysis. Results: Several parameters related to the lipoprotein profile including remnant cholesterol, small LDL and IDL-cholesterol were found to be significantly increased in the dilated group compared to those in the nondilated group. The glycoprotein A-nuclear magnetic resonance (NMR) signal, a novel inflammation biomarker, was also observed to be increased in the dilated group. After performing multivariate analysis, remnant cholesterol remained an independent variable related to AAoD. In the long-term follow-up, proatherogenic lipoprotein parameters were related to ascending aorta dilatation velocity ascending. After a lineal regression analysis, non-HDL particles remained as an independent predictor of ascending aorta dilation velocity. Conclusions: Patients with BAV and AAoD presented a more pro-atherogenic profile assessed by 1H-NMR, especially related to triglyceride-rich lipoproteins. This pro-atherogenic profile seems to contribute to the higher growth rate of ascending aorta diameter.This research was funded by “Proyectos de Investigación Clínica” from the Spanish Society of Cardiology, grant Bayer 2019. BA-G received a research scholarship within the Martí-Franquès Research Fellowship Program from the University of Rovira i Virgili

Different profiles of lipoprotein particles associate various degrees of cardiac involvement in adolescents with morbid obesity

Dyslipidemia secondary to obesity is a risk factor related to cardiovascular disease events, however a pathological conventional lipid profile (CLP) is infrequently found in obese children. The objective is to evaluate the advanced lipoprotein testing (ALT) and its relationship with cardiac changes, metabolic syndrome (MS) and inflammatory markers in a population of morbidly obese adolescents with normal CLP and without type 2 diabetes mellitus, the most common scenario in obese adolescents. Prospective case-control research of 42 morbidly obese adolescents and 25 normal-weight adolescents, whose left ventricle (LV) morphology and function had been assessed. The ALT was obtained by proton nuclear magnetic resonance spectroscopy, and the results were compared according to the degree of cardiac involvement - normal heart, mild LV changes, and severe LV changes (specifically LV remodeling and systolic dysfunction) - and related to inflammation markers [highly-sensitive C-reactive protein and glycoprotein A (GlycA)] and insulin-resistance [homeostatic model assessment for insulin-resistance (HOMA-IR)]. A second analysis was performed to compare our results with the predominant ALT when only body mass index and metabolic syndrome criteria were considered. The three cardiac involvement groups showed significant increases in HOMA-IR, inflammatory markers and ALT ratio LDL-P/HDL-P (40.0 vs. 43.9 vs. 47.1, p 0.012). When only cardiac change groups were considered, differences in small LDL-P (565.0 vs. 625.1 nmol/L, p 0.070), VLDL size and GlycA demonstrated better utility than just traditional risk factors to predict which subjects could present severe LV changes [AUC: 0.79 (95% CI: 0.54-1)]. In the second analysis, an atherosclerotic ALT was detected in morbidly obese subjects, characterized by a significant increase in large VLDL-P, small LDL-P, ratio LDL-P/HDL-P and ratio HDL-TG/HDL-C. Subjects with criteria for MS presented overall worse ALT (specially in triglyceride-enriched particles) and remnant cholesterol values. ALT parameters and GlycA appear to be more reliable indicators of cardiac change severity than traditional CV risk factors. Particularly, the overage of LDL-P compared to HDL-P and the increase in small LDL-P with cholesterol-depleted LDL particles appear to be the key ALT's parameters involved in LV changes. Morbidly obese adolescents show an atherosclerotic ALT and those with MS present worse ALT values

Glycoprotein profile assessed by1h-nmr as a global inflammation marker in patients with HIV infection. A prospective study

Plasma glycoproteins are a composite biomarker of inflammation and can be detected byH-NMR. The aim of this study was to prospectively appraise the clinical value of plasma glycopro-teins assessed byH-NMR in people living with HIV (PLWH). A total of 221 patients with HIV infection were recruited and studied at baseline and at 48 and 144 weeks. Patients were distributed into two groups according to baseline CD4 T-cell number below or above 200 cells/µL. Patients with fewer than 200 cells/µL were distributed into the responders and nonresponders according to antiretroviral therapy (ART) response at 144 weeks. Glycoprotein concentrations were determined byH-NMR arising from the protein bond N-acetylglucosamine and N-acetylgalactosamine signals (GlycA); and N-acetylneuraminic acid signal (GlycB) associated with the sugar-protein bond con-centration and aggregation state (shapes (height/width)). Basal glycoprotein concentrations were higher in patients with < 200 CD4 T-cell/µL (Glyc A: 1040(917.9-1199.1) vs. 950.4(845.5-1050.9), p < 0.001, and Glyc B: 521(440.3-610.3) vs. 468.6(417.9-507.0) µ mol/L, p < 0.001) being reduced by ART. The height/width (H/W) ratio was the parameter showing a better association with this clinical sta-tus. Baseline glycoproteins predict the condition of responder/nonresponder. In this study,H-NMR glycoproteins provide novel insights to assess inflammation status and have prognostic value in PLWH

Free triiodothyronine levels and age influences the metabolic profile and COVID-19 severity parameters in euthyroid and levothyroxine-treated patients

Metabolic reprogramming is required to fight infections and thyroid hormones are key regulators of metabolism. We have analyzed in hospitalized COVID-19 patients: 40 euthyroid and 39 levothyroxine (LT4)-treated patients in the ward and 29 euthyroid and 9 LT4-treated patients in the intensive care unit (ICU), the baseline characteristics, laboratory data, thyroid-stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), the FT3/FT4 ratio, 11 antiviral cytokines and 74 metabolomic parameters. No evidence for significant differences between euthyroid and LT4-treated patients were found in the biochemical, metabolomic and cytokines parameters analyzed. Only TSH (p=0.009) and ferritin (p=0.031) showed significant differences between euthyroid and LT4-treated patients in the ward, and TSH (p=0.044) and FT4 (p=0.012) in the ICU. Accordingly, severity and mortality were similar in euthyroid and LT4-treated patients. On the other hand, FT3 was negatively related to age (p=0.012), independently of sex and body mass index in hospitalized COVID-19 patients. Patients with low FT3 and older age showed a worse prognosis and higher levels of the COVID-19 severity markers IL-6 and IL-10 than patients with high FT3. IL-6 negatively correlated with FT3 (p=0.023) independently of age, body mass index and sex, whereas IL-10 positively associated with age (p=0.035) independently of FT3, body mass index and sex. A metabolomic cluster of 6 parameters defined low FT3 ward patients. Two parameters, esterified cholesterol (p=4.1x10) and small HDL particles (p=6.0x10) correlated with FT3 independently of age, body mass index and sex, whereas 3-hydroxybutyrate (p=0.010), acetone (p=0.076), creatinine (p=0.017) and high-density-lipoprotein (HDL) diameter (p=8.3x10) were associated to FT3 and also to age, with p-values of 0.030, 0.026, 0.017 and 8.3x10, respectively. In conclusion, no significant differences in FT3, cytokines, and metabolomic profile, or in severity and outcome of COVID-19, were found during hospitalization between euthyroid patients and hypothyroid patients treated with LT4. In addition, FT3 and age negatively correlate in COVID-19 patients and parameters that predict poor prognosis were associated with low FT3, and/or with age. A metabolomic cluster indicative of a high ketogenic profile defines non-critical hospitalized patients with low FT3 levels.PID2020-116146RB-I00 from the Ministerio de Ciencia e Innovación with European Regional Development Funds (FEDER), BMD-3724 from the Comunidad de Madrid, 202020E169 from the CSIC, 2020PANDE00082 from the Generalitad de Cataluña and Fundación Hay Esperanza

Determinación de marcas epigenéticas específicas de tejido para la identificación del origen de las vesículas circulantes

El presente Trabajo Final de Máster (TFM) pretende dar respuesta a la necesidad de identificar el origen de las vesículas extracelulares circulantes. Así teniendo en cuenta que las vesículas extracelulares contienen material genético de la célula progenitora, incluyendo ADN genómico, y que cada tipo celular tiene un patrón epigenético específico, el objetivo principal de este TFM es la aplicación de métodos y herramientas bioinformáticas en datos epigenéticos depositados en bases de datos públicos, para el diseño y el desarrollo de un algoritmo clasificador de marcadores epigenéticos capaz de diferenciar entre tejidos. Para alcanzar el objetivo principal se han analizado los datos referentes a la metilación de DNA de un total de 252 experimentos, agrupados en 24 clústers específicos de tejido o tipo celular, y mediante la creación de una función (BiosourceMapper) se han identificado posiciones o loci diferencialmente metilados para el tejido o tipo celular seleccionado, así como las regiones enriquecidas en posiciones diferencialmente metiladas. Además para mejorar la visualización e interpretación de los datos se ha creado la aplicación shiny Biosource Mapper App, que permite poden acceder y analizar los resultados generados durante este TFM a cualquier usuario. La principal conclusión obtenida es que mediante el análisis bioinformático del patrón de metilación con la función BiosourceMapper implementada en la aplicación Biosource Mapper App es posible obtener marcas específicas de tejido.The aim of this Final Master's Project (FMP) is to identify the origin of circulating extracellular vesicles. Taking into account that the extracellular vesicles contain genetic material from the progenitor cell, including genomic DNA, and that each cell type has a specific epigenetic pattern, the main objective of this FMP is the application of bioinformatic methods and tools on epigenetic data deposited in public repositories, for the design and development of an algorithm based on epigenetic marks to differentiate between tissues. To achieve this objective, the DNA methylation data from a total of 252 experiments, grouped in 24 tissuespecific or cell-type clusters, were analyzed and positions or loci were identified by creating a function (BiosourceMapper). Differentially methylated positions specific for the tissue or cell type selected, as well as regions enriched in differentially methylated positions, were found. Furthermore, to improve the visualization and interpretation of the data, the shiny Biosource Mapper App has been created, allowing users to access and analyze the results generated within this TFM. The main conclusion obtained is that bioinformatic analysis of the methylation pattern with the BiosourceMapper function implemented in the Biosource Mapper App identify specific tissue marks.El present Treball Final de Màster (TFM) pretén donar resposta a la necessitat d'identificar l'origen de les vesícules extracel·lulars circulants. Així tenint en compte que les vesícules extracel·lulars contenen material genètic de la cèl·lula progenitora, incloent ADN genómico, i que cada tipus cel·lular té un patró epigenètic específic, l'objectiu principal d'aquest TFM és l'aplicació de mètodes i eines bioinformátiques en dades epigenètiques dipositades en bases de dades públics, per al disseny i el desenvolupament d'un algorisme classificador de marcadors epigenètics capaç de diferenciar entre teixits. Per aconseguir l'objectiu principal s'han analitzat les dades referents a la metilació de DNA d'un total de 252 experiments, agrupats en 24 clústers específics de teixit o tipus cel·lular, i mitjançant la creació d'una funció (BiosourceMapper) s'han identificat posicions o loci diferencialment metilados per al teixit o tipus cel·lular seleccionat, així com les regions enriquides en posicions diferencialment metiladas. A més per millorar la visualització i interpretació de les dades s'ha creat l'aplicació shiny Biosource Mapper App, que permet podin accedir i analitzar els resultats generats durant aquest TFM a qualsevol usuari. La principal conclusió obtinguda és que mitjançant l'anàlisi bioinformático del patró de metilación amb la funció BiosourceMapper implementada en l'aplicació Biosource Mapper App és possible obtenir marques específiques de teixit

Specific circulating microRNA signature of bicuspid aortic valve disease

Filiació URV: SI DOI: 10.1186/s12967-017-1176-x URL: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5387230/Background: We aimed to determine the circulating miRNA expression profile associated with BAV and aortic dilation to provide diagnostic and prognostic biomarkers for BAV and/or aortic dilation. Methods and results: We applied a miRNome-wide microarray approach using plasma samples (n = 24) from healthy tricuspid aortic valve individuals, BAV patients and BAV patients with aortic dilation to compare and identify the specific miRNAs associated with BAV and aortic dilation. In a second stage, the expression patterns of the miRNA candidates were validated by RT-qPCR in an independent cohort (n = 43). The miRNA microarray data and RT-qPCR analyses revealed that the expression levels of circulating miR-122, miR-130a and miR-486 are significantly influenced by the morphology of the aortic valve (bicuspid/tricuspid) and could be functionally involved in the regulation of TGF-β1 signalling. Furthermore, the expression pattern of miR-718 in the plasma was strongly influenced by dilation of the ascending aorta. miR-718 expression was inversely correlated with the aortic diameter (R = -0.63, p = 3.1 × 10-5) and was an independent predictor of aortic dilation (β = -0.41, p = 0.022). The genes targeted by miR-718 are involved in the regulation of vascular remodelling. Conclusions: We propose that miR-122, miR-130a, miR-486 and miR-718 are new molecular features associated with BAV and aortic dilation principally by the activation of TGF-β1 pathway and vascular remodelling mediated by VEGF signalling pathways

Data on the circulating levels of endothelial microparticles are elevated in patients with bicuspid aortic valve and are related to aortic dilation

The data included here support the research article “Circulating endothelial microparticles are elevated in bicuspid aortic valve (BAV) disease and related to aortic dilation” (Alegret et al., 2016 [1]) where circulating levels of platelet endothelial cell adhesion molecule (PECAM+) endothelial microparticles (EMPs) were identified as a biological variable related to aortic dilation in patients with BAV disease. The data presented in this article are composed by four tables and one figure containing the clinical and echocardiographic characteristics of the patients (Alegret et al., 2016 [1]) included in this study, and summarize the results of multivariate linear analyses. Furthermore, is also included a figure showing a representative flow cytometry dot plots and histograms used in PECAM+ EMPs quantification is also included